c - HPS Repository

-I-
YOL
Y CO '
• B llard, 1rofessor of
•
onald
• . ostello,
iology,
1ofessor of
ool
y , in
in ton ln
,liktor Hamburger, J-11of s .. or of Zoolo y,
c rge of co rse •
rt
ut
Goll
y, Univetsi y of
1
rth C ro-
lina.
• O~penheimer, Asslst nt
College.
J ne
·ol
m.A. ~ c,u.:.r. f'.t~Uc-u
E.e~~ J;~,
{'~~ ~
rof s or in
/
-s
~~,.AA. ~~J ~ ~
,
r
-).-
bryology Co· rse
.roods Hole ,
1945
Beach, Janet - U. of Conn.
Bernier, Germaine A. Berry, Beth SinclairiPyaB:t, Ltmi enee 0.
Carter,
:arjorie E.
u. of ontreal - (Instructor)
~ockford College l(g ~ ~
= U.
of Fa.
Ga. State \'omans College - ( B.S., College
of iilliam and Mary ; 1 • .ft ., U . of Va.)
Chirico, Anna I!arie - Seton Hill College
Clark, Carl Cyrus - Amherst College - (.u.s., .. orcester Polytech. Inst.
Copinger, Anne Stevens - Goucher College
.t!.ihrlich,
uri am
-
Knox College - (A. B., Knox College)
Izzo, lary Jane- U. of Rochester
Kell, Arn:y - Central Col:.ege, University of Ill. - (.A . B., Central Col.)
Levin, Ilene Blanche - Goucher College
Lodico, Dorothy G. -
u.
of Rochester
Lovelace, Lollie Roberta-
u.
of N.C.- (B.S., U. of S.C.; M•• ,
U. of S.Co)
&arker, lviuriel Josephine - Colby College ( f\. l5 , t2-oL.. I'
y
Mezger, Liselotte - Bryn IJ1awr College
1liller, Helma Catherine - The Johns Hopkins U.
Perkins, Barbara Burnham-
u. of
Conn.- (~ -
s.- '4- o\ Co~C\)
Raymond, Barbara - Swarthmore College
Rice, Hary Esther - Drew
Roberts, Elizabeth
s. -
u.
U. of Pa., (A. B., Dickinson College)
Ruderman, Claire- U. of Rochester- (A. B., Hunter College)
Thorby, Jean Adelaide -Rockford College
Uphoff, Delta Emma- U. of Rochester- (A. B., Russell Sage College)
-J-
Tentative Schedule
bryology Course, :oods Hole, 1945
Thu .
Fri.
Sat.
Sun.
..on.
Tue.
v·ed.
Thu.
Fri.
Sat.
Sun.
~
11
oon)
e~is)
. e~is)
ew :rv;oon)
Mon .
Tue.
v~e d.
Tb.u.
:E'ri.
Sat.
Sun.
I..on.
'fue;.
VIed.
Thu.
Fri.
Sat.
Sun.
Mon.
Tue.
Wed .
Thu.
Fri .
Sat .
Sun.
Mon.
Tue.
Wed.
Thu.
Fri.
J"une
June
J"une
June
J"une
Jun.e
June
June
June
June
J"une
June
June
June
June
J"une
J"une
July
July
July
J"uly
July
J"uly
July
J"uly
J"uly
July
July
J"uly
July
July
July
July
J"uly
July
J"uly
J"uly
14
15
16
17
18
19
20
21
22
23
Squid
Squid
Squid
V.H,~
V.H.
V.H.
Teleosts
Teleosts
Teleosts
Teleosts
Coelenterates
Tunicates
J.O
J.O.
J".O.
J.O.
chinoderms
chinoderms
chinoderms
V.H.
V.H.
V.H.
B• Bi t4f'ttt..
B.B .
24
25
26
27
28
29
30
1
2
~~~~~~F (~I
I
Fertilization
t\
D.C.
c.
~P't;llj~'!ft!"~~----~• .
3
4
5
6
7
8
9
Cell lineage
Cell lineage
Class Exp. Echinod.
Class ~xp. Echinod.
Annelids
Annelids
D.C.
D.C.
V.H.
V.H.
D.C.
D.C.
Mollusca
10
1 ollusca
11
Coelenterates
Tunicata
J.O.
J".O.
B.B.
B.B.
Class Exp.
Staff'
12
13
14
15
16
17
18
1920 ·
.
e-~
-~ -
RYOLOG . CO
SE
1945
Exper· ental Period
Group I
_,xial gradients and :Ph. siolo . . ical dominance
Tubularia - Dr. Ballard
n
Berry, Chirico, Izzo, Kell, Levin
Grouu II
Control of Polarity in Tubularia -Dr. Ballard
Carter, Perkins,
Grouu III
arker, Rice
Isolation of blastomeres in !Toreis - Dr. Costello
Ehrlich, Lovelace, Raymond, Uphoff
Group IV
Isolation of blastaneres in IIydractinia . Polar lobe
extirpation in Ilyanassa - Dr. Oppenht::ime r
Beach, Copinger, Lodico, 1illeri Roberts
Group V
Centrifugation and other experiments on
Dr. Hamburger
Clark, Eezger, Ruderman, Thorby
'chinoderms -
-sEY.BRYOLOGY COURSE 1945
Lectu1~s
Thursday, June 14 9:00 A•. l .
Embryology of the Squid I
Dr . Tamburger
Friday, June 14, 9:00 A.M.
Embryology of the Squid II
Dr. Hamburger
Saturday, June 15, 9:00 A.M.
Reprodurtion and Spermatophore
of the Squid
Dr. l-famburger
Monday, June 18, 9:00 A. M.
Embryology of Teleosts I
Dr. Oppenheimer
Tuesday., June 19, 9:00 A. M.
Embryology of Teleosts II
Dr. 0}-penheimer
Wednesday, June 20, 9:00 A.M. Gastrulation in Vertebrates
Dr. Oppenheimer
Wednesday, June 20, 8:00 P . M. Seminar:
Thursday, June 21
No Lecture
Embryonic Induction
Dr. Hamburger
-ID BRYOLOGY COURSE, 1945
LECTURE::>
l<'riday, June 22, 9:00 a.m.
Embryology of Coelenterates
Dr. Ballard
Saturday, June 23, 9:00 a.m.
Embr1ology of ~tyela
(Tunicates)-Dr. Ball
Monday, June 25, 9:00 a.m.
Embryology of Echinoderms
Dr. Hamburger
Tuesday, June 26 1 9:00 a.m.
No Lecture
Wednesday, June 27, 9:00 a.m.
Experimental Embryology of
Echinoderms -Dr. Hamburger
Wednesday, June 27, 8:00p.m.
Seminar: Organization of the
Teleost Blastoderm
Dr. Oppenheimer
- 7y
r~d
r
y,
0;.8, 9•
y, Ju
:n
29, 9:00
J
y
5
• •
• •
J.c.C
/J .v~.
.
30
y 2,
:0
• •
c
•
1
0
-a-
Cour
bryolo
u s
y, July
r ,
:0
•
rt
•
, 1
no
• rtiliz tio
n ni
r . Co t llo
edna day,
uly
o L ctur
th,
Thursday, July 5tl,
~ : 00
xp
••
n
for.
th
111 r,
hn Hop
in
Di"CU ~ o o
g noo in d velop nt
8 : 00 p . m.
~
Dr . Cost llo , Dr.
Frid y , July 6th ,
Developn nt of
9 : 00 a . m.
Dr . vOStello
Saturday , July 7th
ro es'or
n
n1v ro ty
.,
No Le c tur
rolo
nbu:r er, Dr .
nnelids
f
1111 r
-.u..ffiRYOLOGY CO "'RSE
1945
Lectures
.onday, July 9th
9:00a.m •
Embryolo y of •~allusca
Dr. O:p . . enhe imer
Tuesday, July lOth
9:00a.m.
Ex:perimen ta 1 ..!.mbryoloc-y of
nnelida and .ollusca
Dr. Costello
Pfednesday, July 11th
9:00 a.m.
Problems in Coelenterate Developmen
Dr. Ballard
Thussday, July 12th
8:00 pm.
Seminar: ~etabolism and Development
Dr. E. J. Beall, Assoc. Professor
of Zoology, Yale University
9:00a.m.
Asexual Reproduction in Tunicates
Dr. Ballard
Friday, July 13th
No Lecture
Saturday, July 14th
No Lecture
- ;o -
LECTURES
1945
1 onday, July 16th
No lecture
TUesday, July 17th - 9 : 00 a .n! .
Expc1· ntn
t des of the
different at o of tl
car eetod n:n
in the salamander.
Dr . C. L. Yntema, Asst. Professor of
Ana omy, Co ell Univ rs ty edlcal o h
New York
lednesday, July 18th - a .m.
No lecture
8 : 00p . m. Seminar : The influe1 e oft e nervo,
syst m .n re enerat: on 1
h
1 •on le
amphib ns
D • ¥arcus ~ n er. Asst . in Anato:ny,
I nrvard Medical :::>chool
Thursday, July 19th
Friday, .July 2J th - 9 :Oo a.m.)
2l 00 p . m. )
o le t re
Repore of students on the r exverim nts.
Report of Students on Group Ex eriments
Friday, July 20th
g:OO a.m. {1) Isolation of blastomeres in Fydractinia
Miss l\·iller
{2)
1ethods for isolating blastomeres in Nero is
.iss U hoff
(3) Development of centrifu ed eg s and egg
fragments of rbacia
iss Ruderman
(4) Effects of lithium chloride and alcohol
solutions on the development of Fundulus.
l\iss Haymond
(5) Disintegration gradients and oxidat on Reduction gradients in Edr inoderms
Mr. Clark
(6) Axial gradient in
re~1lation
rate of Tubularia
Miss ~ arker
2:00 p.m. (7) The influence of dominance on reginera tion in
Tubularia
Miss Carter
(8) External factors in regeneration of Tubularia
lVlis s Levin
3:00 p .m. Demonstrations
~-
1944
D,v~lo~~e~t
of tte m2leost
1.....::...• t~·::-es of ee,c;s v:ill usuclly be st dj.e<i;
non - vela<.:ic ee, .... .
• The non-)elaGic egg.
E.~~.::.:-:l)les:
Fundt,_lus het~;rocli tus
FUl dulus L'" j 2lis
Opsanus tcu (Lhe toad fish)
Fist arc usunlly broug~1t
AJ. ter strip _>Ll[
-.1 e"'isferred to a discard tnEk.
I.t' netericl
·-c-, keep :.1ales and fe;::ales i rr s.3p rrota tsnks
·.P. lE-ss likely to s~lcd their ee_:,; s.
J. .
CAR3 OF Fls:I •
J :,t,s of w::les 2nd fe;y les .
to
of
is
cs
t,
e 9elc.:..ic
g: end tl1e
the lC'bor::- t :- r • in r::ix: d
eggs end sy .rms they nre
lirui te it ~ . s bot ter
t1e:: f-3I:ltlcs \'J ill then
G. F . ...c..?A?.A.~.'IOE JF CU.!../EJ'"RES . '.I".-0 cultures s:!ould be str.rtod by either
.stud..Jnt or instructor {!.uw)'...!IlCu.nent \'ill b<; !:'!c.de) n the first dC\y,
n~a at 10 a .n. and tla second ns late in th0 avenin[ rs prac~icnhlu.
P.rJ·:-.n~lJrt;!;ts should also be :~led.:: nt eac~1 tDble to stcrt et lot st one
cul tu1 e for ir.r..:0d1:1te study of fertilization .
a) Strip 0g,gs into a clean bowl which :1as bG·-D ~.:oistened b" sea
wu.t.sr . Stri.J r.:ilt and. nix v•itll egc,s v.rith bc1oly suffici,:;nt salt wrt .... r
to cover the eggs. Eg,__ s s~1ould not be E'llowed to stf'nd :.:ore t 1an 5
r:~inut0 s '0a for~ for til i zing.
After ten r.:linu-ccs add e.nd chancE) t ·eo wt1tc:r
and lGava in shallow wct~r (not ~ore than l/4 inch deep). Ke ~ ) bowl
cov0rcd ni th glass :plato . Do no t at an.' tir.~e allo\: eg~ s to ·!lump
or accur.1ulate in ~s~
Label oach lot vi th exact ti:ile of i:ertili~ation.
Chango watar at le8st once a doy .
b) The eggs should first be studied in the condition in whioh thoy
are sp awned llut for ronny purposes it is dE>sir·r-blv to :_::>re;pcre the ·l€;[ s
as follows for ~icroscopic study : roll t~e; egcs on a piece of fi~ter
pfiper until the jally end outer fibres ~re ronovsd le rvinG t~o surface
of tae o·J.tCjr Ll3ra.breno smooth e.nd cl·J an . Pl e ca in sea v1at~r in c ul turo
slide in ,·,•lnch ti1e d.;;pth of tha depres s1. on is slightly los£ tr1~n tlle
diam:;t·.::r of the egg .
( Thssa c.re prov1dcd in t:1e lflboratory equipment)
Th;:; eg[. may now be rotc.tad by moving the cover slip. Another ..'!!e thod is
to count on an o ~d in e ry [l~ss slide in sea watar unda r a thin flexible
pit;C0 of r:uca peeled fro:.:-11 the thicke;r mica sheets thf.1t are provided.
Drew off the wator until capillary attraction caus~s a pressure on the
cge, and rotate it as 1n the :precedin€:, method .
T".till OVUl.:.
To study unf e5r tilized ogt..s they should be obtained from
a fom.ale tl1at hes been k0pt in fresh wr- ter for about 20 minutes. Strip
nnd keE-p the egg in dilutt; su1 n:.ter ( ?0'/o fresh wat;;:r, 30% sea water)
to ratnin the norphological chAractoristids observP..le at time of
extrusion. Note details of structures of th...; unf :rtilized mAture ovun.
l~1GSO include yoli~ plntes , oil drops, protoples1:1, !,er.lbrAnes, ni crop yle (
etc . (The micropyle ::1ust b0 ob3c.rved before r~mov('l of c.aorionic jelly J
If ilJ... .. c...ture ova llr8 pre:s::::nt cor:..parc tllbSG r•ith m:'ture ove.
3.
4. FERTILIZ.t_ri'IO:?. !Jet·.; .:::xect ti.:-.1e of fertilizetion and be _yrepr:rGd to
::;r:.udy i:o.c!•:;dioto c :!ant.'-'s . ~Jotc ti:.1c of' c .:.ta:l(..O of yolk plrtcs; of formetion of p0rivit0llin0 spaco. ~~ot rr~ your conclusions in r ~~rrd to th
r n, idity of the activation of tho ec:L::? If ~) rrcticebla find :".1icro:,>Yle
on u.nfurtiliz..:.. d ee;g . InscunnrtG ond ~~0t" s:Ho3Cdi!l(. of fert1.lization
- 2·r:. c.ctio~~
fro •.: l.:>cus to r::1.cropyle .
. .t.1 .
.
1:ot. .... th..; r ')du£ l c;::c ;-.ulo c1o~1 of
--- c::· ~·....::.. !3.L.C .::.':22DISC
. ........
.- ro'to?L.J :..:..1c ~t:
hl.s is t:1s bla~+:oJi.sc ,,r (!~r:·.~ disc .
Co.c::pl'ro
pD r ::na. l"t..:.r·-"·1. \'i-.::~ =s .
?ol:r bodi-:s r..r'y rw ·,.):·. e vm t< l.-)usl:· st:.ldied
ii.. G.~-: t ::.c ::r.~s.
.Jwt is th-: r...:lrtt. i n '"'f tl.-.:. }>Ol. of t .ll-.. ovura to rr ~;:ity'? :rov: .:10'-'s ~.is c·~p3::: \'lch t:1· condL\i~on ~n t:!u frol. .:;Az; \'itl
tl.C C.'.iCk it;g? :i)q f.., r •• rOC .~S t...:S i-f.;(:., 'Jl~CD 1!1 t~h. U ...1i'Crtili20d .JL
::;1Lil :.r tc c:::>s..) .:.r:. t. t ... f._;:··~iliz~.-d c'=.c?
· ,
, . ..., _ 1 .....
~T_}'AVAGE .
·;; .:tc.:.i for t:L r-:1.:? ;rJ c::;nct: of - [-roov2 1n tho surf~c0 o ...
T:.is t.;.SU (>lly
3 bl<:.,s toC..i.sk-t~12 it dicc.tior: o"' l-:1s f:i.I·f; ~ c)"' cv ~ o )ltn.-3 .
.c·__ .,. fros t\:o co t·1r0~ ~o1~1s <-'l~c-:r :'-...rtilJ..~~ tioT' . '.D~~ r·t~.. of
.V-.~0pi~·...:i1-'u Vf.ri~t ri't:;:· t r.: t'"'.:~y.:rr:turc •
. Joto til._. [...;OI":.o.;.t!'lC f''i1d 'ti~~O
. , Jl<: ~-ions ·. n' b.• ..; sut,s~(:ucnt cle: ··· nt..os . Do th..; cl...:av:.1~.,; p lr:10s di vido
..
<
·.,
~•
-tirJ ovuru.?
'Th." -..!ltl..:'v
bl'"~..:todi~c?
:Jt.u'i"1~.
~r
t3r}dn·... Slf. t
.:~
nuc-
. J. :tl'·::. so::.---·tl· .. :...s v1sibl--- .
Dist · ::cc1ish t,:;t•. ·o0n eo!1tr ..:J. ::md : rr.:;1nol
C~llsA
F~llm; c:::..cr.vae;;c c~ ... \-f ull~r to t .. L; 32-~~21;. stc r;~ .
rotc
" l'• ·J(-'ll.::r itL·~>s .
,,.hv:'t do nori7o,lt, 1 cl.:..r.v.:- (S.: pl 'J.10S f.~.J'st C.'YPC r? Do,
~~~~
.nstodu c inol'8aso i'1 siz~ or· ~1t0r .t n for:'l?
s
;; 1
· i..ht.t cl.._rvo,-L;f' COiltiP'J.e for r. C:).~ :"id.:rc1JJ.,:; "'t-riod ~··it 0' 1 t
cl·- r. .:;;v ir~ forL fro;:.. t ·1 t of tll-:. or1e;in ·1 blc stod.::.sc . This ir.
c<llvd ti.t3 )0I'io6. c.f ·~_t(. hi,~.. lnstulr. ( S::>.j 0::>, onllvira...:J' ' .30 for
('~•TO.l1·'JlO.J.C'1l t..)'Cds) .
\.'!.;;;n c:u . . s tl.'-' ch.:nge of for::1 to tlh; "~lrb:'
\::::...cs bulu occur ?
~oL.,
l~l,Ch
i:-ot.o : As clevGlOpL.e:ntcl r Pi'.G v ...::r.i.,s ·.·:it1 t~:.:; tJ "l)Gro ·t.ure: no t.-::10 C.'rolJ.,_.lot;y C£..1 be CiVvn for st:Jf:.,OS 01· funriulus fS 0..;G-..TV0d t:nd .r v r:~'rint:.
t0rperut.ur·:;s conclitions . For· list of sto.€_.,s s0 ·J :;c,p...Jl' by 0?~)Cl1h )if."',c r
' 3?; for c:n·onolu~2 at f:50C s •. c :;;:>Lpor by Solo~rg ' 38)
7.
T.~.S l):!:J.Il..L '.ST .
( First L )!.)3<I'8ncc is fro·~ lu to .::Ll .... ours aftt;r
f-,rti li2.ction) .. 'Eo unc 1--- :;v ..;C:.. .)r :.'to:pl:-.sL ,.:: ounJ t:-.. ..:: .HJ.r [ i '1 of tho
( ):OU) of bl0sto1nurcs is Cfll lcd the :s2ort; in -~ l -:::>.:..rlbl<st; t:v· ~ 'uc.n...Jn'Ch
t~~ blesto~~sc (not visibl~ 0xcapt in sections) ; thv c~ntr2l per~plrst .
In t1.,:; le.t i) blestodlsc (18 - 20 hours) obs0rvo particulorly t!~..; boh · vior
of thw ~a r ginel cull s snd distinguish bc.tv·~~n circul~r ~nd ~~dirl cl~c­
vnGos . T~1.;. lr-rt;;o pi{lkir:ll nuc l .:::i of tno :-.:;;riblcst cro ;Psily visible .
Not: !1m tn...; nucl0i of tlh.. : .:;..s.re:.inr.l ro' · of c .:.l ls b::..cor 0 fr,;...; froi·11 Cl:.'ll
o~ttlin·.:.s , contLLlv t .(ijL divis1ons cnQ !;li ( r ... t -~ into t~~c rM'l'( ::.n l :;cri.l'Jlr st , conv~n tin ~,:; it i::Y~o a nucl.n:tcd out non- cellulsr struoturo .
Note t .-:. contir.'J.;-tion of' thG ::~cr:L bl3st .s tructurc :;.n l · tc.r stre::es .
8.
T:IE G....a · _ RHW .">LD .c:.:....: EX'l\;,l'(S.iOl~ 01!" J.':i~~ BL\S 1'GDISC .
( 18 to 48 hrs )
s .~lic::3Q.'10 ~ltl~' to tl1. 11'10L:rtion of the ::;>oriplcst note; tllc C.•.lPng0 1n forl'l
;.:~dJ. siz ... of' t:.z, 'btr.oto(h,r:-:. ( tlrst'..llc ).
Soon t11e mcrgin of tL0 disc
'i 1p0<1rs rol ::;'tively thich:Gr ( corm. i'ing ) d·l.tv both to rr1 :·ct·1.. 1. tr.lclcvnitlb c:.:'lO. GO L•• i.. t~l.:.lillin , of tli.C C..; ntrrl prrt Of th3 disc .
I''i.c t..~rl<J.
riq<: cr.:1 best b..:. olw;rvv cl in :F • ..•~jclis . Durlnf. ti.1e n.,xt f .. ' hours
t 'l..:; L_,i:. I' •. ~ ri!'.t_ fi'O\JS C0..:1plvb0ly OVOI t:~~ SUlf'•CO Of th: :·olk r.lr S" •
'rne uncov·~rod ::~o ... tion o'f' t·.h:; t:.L3 is th,; blr.stoporc . Tho fin e l oov~rinc
of tl~c; 'Tol~ or t _,c clo:~ur .- of th..;; blc stopor0 occurc nftcr tllc f1r st
s C0cGS or t~l, for ~-:: .'. 1,lon of tl• '~ 0.n~,r: 0 . Undvr f"vor f'blc. c::>nd \ t ions tbo
·J.:;t.ln .lllf:- of ['[ .strul tion :..··:.· ~:8 obscrvJd in the ~p')O<I nncc of r:
... , ight ind·.3:'1tr.tion 1 t t~lC.. uci['G o' t;IlC. [,wr~l r-L1' ,.,t n t.t.I .i.i '' 1!vll t!1•J
vo.1.k is cb0:1t l / 4 cov0r0d . Strinint- VJit.h n ... utr"l rud ury ·.:e l'!) in
- 3:d0n~ific ction
c-
of
~~ru ri~ 0 •
(Usc 1
r 2 dro s tor
s·r~cu~~
d1sh of
:,s. , ·-t.:r) .
24 tc 36 hrs . )
9.
. . 1·1 ..;- tvi!
, _ ...._ l...:.J.
- -·•.• r ·-·-.;~"r
J.C::O
........ ,. ..... " ... 'J.r.
n--o·n
,...,_ ••
(; ··o1'
) .,.....
.... 'JAI..I..i'!U
u:_
-~·
•• 1.1
,·
A, .._v .. \.;; r"nr•"·'•tJ."on
f t~0 o~ ryo tr~es J1cca . I ts first in'ic~tlon is ~ co11t1"r thic~on­
::.n[; :~1:0··1 :-s ti~.... o:....br c.nic s .. L..,l~. CC'.lsed by :' 1010 o tiv": :..ovc::1.... nt of
c 01 ~s L o.:..... K.:'t of tl1..., e>...:r;·. r1n'- .
.J .
T:i:1is for.~.~ ti0:1 .:s 1.ni ti ·· c.:.d v:;"~n t:.-J '1 t' stod .... :rr.:. •• -- s ~ovcr0d fr
.. ·..lt--:rt,r to ..'!~c t..:irC'. of' t: .... ~'..lrf-:::e o:' tl10 :·ol.c
By t .. w t.i:. c t·",t
•·: .: ·)lr. stod,_,r·~ ~· s cov..:r.:d ....... out on"' h.:1f of t~".c :·ol·;: t::... ~.,:..br:;onic
: ; 11 ;:~c. ::.:;s bc..co::c r ·)l''"ltl:· tric.<_,ulr>.r rr0c ext.JndinL~ fror. the r.vJrGin
~ .:· )?'lt- ~or·t.ioi.: of tlL blcstodar;·.: to n-.. r tl.c center of the blnstod0r::1
: :1 :. .;,;.tr:jonic Slli..:::ld c,...:1 1:1ost r ·:::P ~ .tl:· b-J id-: :1t.:..fL::d r•:: n s ... un L~ profi1o •
..s ~::e: bl-.stod..:.E.; s::;>r.::~'ds ov.:r t.J:i:-; su:ftc-:: of the. ·oL~ tl-:.: u::.b: ;·"' t7 .. 0'"s
T<:)i.d .ly i.li 1-.:.me..,tr• •
'!'3
i) . ) O.:s..:..rvo th•3 i.'holo O£.[ in prof.llo vi.Jv, so thct. ti1"" :...r:1br.,ro is
.::.J~l in ..,r.bi tt r<i. opticc::l section , ~·,.•~n t l o yolk is 1/3, 1/2, B/3 ond 3/4
vOV-:;TLd . \::~r- t :noportion QOCS tllG l._;ngt.:l of tlKl c:.:br:·o ba, r 'tO the
di "''-'-'tG:r of the blcstod.J:rr.1 rnd to t :.:; 1Gn£ til of t~:..; .>I : r11:.. in .... c ••
,.
cf thLS-
s~ccessivc
steges?
c . ) Aft ...,r tho yolk is 7/8 cov(:r ·~ d, look for ~ lrr·..; cl.:Jcr vositliO hind end of T-lw onbr~·o . (d o not confus.:: this \"it f'l
c1·1s~ .... r of s::inll oil dr-o:ps frcq_uentl:' fo•u<i L.1 P. sh.ulrr nosition) .
r,n ·_s is K..1_rfft::r ' s vesicle.
cl c
~1vf
d .) Th€- vi",lbr:-·o ;ys b 0cor:.... s,,6mont,;d . This sogi,:Jntction is confinL.d
to tlle; m~sod..;yn v:hlc h lias on :::rc:1 sido of th3 r.xis ofthc <1i:.lbryo for:· ~ ing
L.osobl2.stic soJ.:it;;s . Hov• l.~.::-ny sonit.es do :·rou find ot th0 tin;.; of t 11....,
closur-a of tha blastoporo ?
5), Lo .Jk for tho notochord . Study it in lont:)t.;.dinsl t:nd trrnsvarsc opt .i.e e l s0c tions . \·.~1Gr.J doo s 'it t ornin rt...: ent~riorily rmd post
criorly?
10.
E:r.. d
L t I':..::t: D...Y~LO?.l :Z~~T .
Obtcin s0rl0 s of 0tltr··ros of 2, 0, 4 , 5 , 6 drys
d...;t :-:1lod cor.rp:::r::~tiv\..: study cov..;;rin{ th-.; follO\,'inc ·,?oir~.ts .
e) Sor::itos . l\bt..; fi1."'st epp0ornnce e.'.d det., rmi!10 nulilb,,rs forr.:ed
en the; s·-.1cccssivc.; deys.
b). r h t.. . br;·in C"J.cl S.Jnsc ore:,cns . Find ln 6"I'l~r str . ....:s the
:)9tic v .:;s~.clo0s , 1 -::;:ls , nc~tromercs, n id-for-:; - cnd hL1d brf'in r-:;g.:.ons r.:nd
Lr-~ c.:.. tlltl d-:;v8lo:;:::7le-nt into curcbrun, O!jtic lobes , c.:rc. ' GllUJ.."'. , r.~odullc. ,
otc .
( DLJ\!incs c t 24 hours intdrv t ls \.:ill t i v:; c< c ood r .:,cord of
C..;:;vul.:J) ..lOllt of :;-r_ ts .) Stud_ .. the; d-:;velo]ne:lt of Lie ey0 , olf"ctor~r pit
ror.d otocyst . Hov· is -cl~o lens of the oye for r1od?
i".1C.l-.c
'-'
1944
T.tis b l. -... l· o:;:. r cp: ...: co;1t0ir~s onlv <:- fey; f t.:e ol' r ')"'urs . (F r
r:; f:.r-t:..-:: ces s~., c. £31 or 1 ,. rks f\nd bi li.::l(.rt-:_).!i0s i:1 prpers
f:.t. r: ---~t.--- . )
-; CTv
C(~:'J l c ·c.;:;
I.
--Ge-:n0r
- -l-..-eference
---
r;ort.. s
·?.!':::.c:~st ,
19.35 Tr::: 1 t~ J.t.., ... bl·::oloeie des v ert.!:u r~s.
2n e •
lS Jo ( e-.iitor) Hrn ... uch C...,r vor.l-2:icho <len unu 3.· ..,ri ~~--:ltallen ~:rtw:.c,;:lu .. u8lt.:.J.l't; u.-..r ',;ii·bt:l t1~re.
J t)nr:. ·
Je ~:-L:so:r. , J . \;. 1913 Ve-rt..:. r e te e:!br:·olog:' . 0).; .;::: c. , Cl )ren · ~!1 Prccs
·,~r::-: , ,T.G,
1 Sl9 TextboJic o:' E·~br~oloe_y, Vol . II, Vertebrrtes, 1., c
c:_ 11 n..ii C,: •
;,~0~ :->..;:r. , T.H .
192? Expsri:uentel Embr,rology, r ew Vork .
~- ,"J')3nh.&i.:: ..:::r , J.:.J. .
1906 Histor1c e1 l~troduction to tle study of
talaost~cn ~evelOpDant.
0~~liS 1 Z, 1936.
s-: .l~i _'~ , 'ii. 1929 . Di0 dct.;;r .. ~in.r-tion der Pri:.. it.ivenC\'!ic.dunt., L .,;.;zit: .
~5ecler , H. E .
1902 L~ h ~buc h aer v ergl . E~ -~~i c k1 un gsgaschichte dor
niedcr e n \,'~r bel tiE.re .
A.
-~--rt ··ib, 0.
li ._S...::_.;:ci~t l
g<l'3iZ , A . c.:d. \,'hl t •.~nn
R..;..:::rences on_Descri· tive E :.hr.ro loe:y
,
C.O. 1884 On the cleve1o_:"lnunt of so:·."e
Pt:lat;ic Fisl1 eg g s. Proc. A ..• Av -.. C . Arts
Science s , 20 •
.. --------------------------- 1885 T::.e d;:,ve1o?::-Jent of Oss~ous F.i.~ :1es .
Iviem. Lus. C on.~ . Zool. C2.::1brid. ._ e , r.:f:\ss . 14.
_(.:._, scr ong , P . B. lS36 Ft.:c:.~-·JJ iS L of li::- tchi nt. in Fundulus hete roclitus.
B... o. Bull. 71.
Cl£.)~, C. I,i .
l 6S l
SoLe _p0i11ts 1:1 the ueve1o~r.2.1t o:;: t:te tord-fish ,
Jou:r. Lor_;.'ho lo~y, 5.
---------- 1 39£
Ti1e leterPl li n~ s yst e t. of Bntrf1chus teu.
Jour.
Uor?i ., 15
Cur:. , l ~lL t!:'r. , J. T .
1385 on t..c r c l r t L:c~ c of t::..e yolk t o th E:: [ .. strulr
in T.. l0:)ster:r.s E .~d i n other Verte br Pte type s. "- ·J. 1· .s., 6.
His, ·,; . 1878 U.:t."'r suchunLcn l!l ·~er di.:~ BilC:ung des K::1oc henf isch0 ...br :ro s.
Arc 'r!. Aaa t. u. Entr:.
Ju r...c s, R. ~ ; . 1939 Ar.£: l :· sls of the devclopr. a n t of fish er:.br~ros by L:t. ens
of t~~ citotic i ~ tiex V.
~1e processes of er rl y d.ifidrentl~t­
l. Ofl of orgPns in Fundwlus l1e reroc1i tu s , Tr r-,n s. Art':. . J,:ic . So c.
58 .
Rpo s 3n -Runce , E.C. 1938 OH th e GPJly devt)lopment, bipol e r difforenti~'-­
tion e ...1G. c,l c;r v c"t;e of t..le :Z.ebr [l flsh , Broc hydc:r1 io revio.
Bio .
B,ll . vol. 75.
R~dar , J.A.
1 832 A cont1·ibUG1o n to the e ~ .r~ology of os s eous fis ~e s
with s~eciel r3f~rence to the cocl. ReDort of Con . of Fish
'
snc.: F :.. si10r .Le s. 1.:r-·::.:.,:nt_ton.
1884.
Sur.m..::r , F.B. 190 .) K:.:.,,1.'f0r ' s Vesicle: rncl its r.:;l rtion to rP.s tru1 rtion
e.nC.. c j:: cr ~sE:.l C3. k0ill . N .Y. Ac ed . Sci., 2.
----------- 1£')4 A .stuciy oi ecrly f1s h dev s 1o -~Jlr.8nt, 3X:!)crii7.e.ntPJ r nd
r::or-p:wlc>t_ .!..cr:l .
Arch. EntvJ . 17. 92-ll S ,
S;:.C.c:\:eru. , C.R. 1Sl5 A stuC..:· of v·aadorln e; r.~ese.10:1yr..:r l cal ls on t li. e
livE<, ~'OL\: s a c. .A:.--:. . Jour. An a t. , 18
,,-ils:>n , H . V. 1 891 T.. -3 Gr:tbr:·olo['y of t he Soe B, ss ( Serr enus otrarius)
Bvll. U • .S. Fis .• Com., 9.
-
- 2III . Stege Serias •
Opp0!1.I".LGL.or, J . ..: . 1937 Th~ :1or~.2l strt;e~ of F:.c~- lts !.ct0roclutus .
Arret . R· c ., 68 .
pelluet, D . 194'± Cr1teric fort:.<:; r-..:co .:_!.:t io1 of ~v vclo:"l::.e:t"l st ~es
in ~ha SElr..on ,
(SeL.tO S '"'lr r) . J • •.vr.)h., 74.·
Pric . . . , J . ·.:· , :934 Tne 2:·. ryology of t:·i;;; ".ita fish, Cor0t.:,i)nus cl perforr:lis . r . rr . Ol.io,J . Sci., vol. ... 4 .
--- ---- -- -- 1935 sm·:e , III . ibid . , vol . 35 .
solbcre, , A. N. 1938 Tile deve:lo:·lEt:nt o1 <' bonv fish (Fu .. d 1us) Pr · t,r.
Fisa CU.:.turist , rn Do.:.,.t of C r:... .• BuraRu of Fisl'• • , '.:•. sh.D.C.
bu.r 6 er , J . \; , 1~31 Soce ex:podrli..vnts on t: v r~;;1:'tion of the ~.:xtc:rni':l
Gnvlrom:l·3nt t-:> t1:-3 sn::!'.:1Pto.' ~n .... tic c ·cla ,) - FunC:u1us
h~tvroclltus .
Biol. B~ll , vol . 77.
C' Teie;-Bennet
l£01 A. T.. c. re;)roductiv.:;, c:·cltj of the threc-soi1wd
s tric lo brc.l{ , G< s tz:rosteurous acu1artus, Ll.u •• • P ••. _. Tr :,:.s .
F. .... J·. Soc , Lvu:.. . B , ...,1£' .
he·., ..ar.n , N .. H .
1., )7 Sprv·ning behe:vior end s.;; xual dh1Jl'pllis:::-u of Fund t1us
hst~roclutus End rl1ied fish .
Biol . Bull. , vo1 . 12 .
•
J.T.cS"~::one:;,
..L
.......
"'
l ., vl F''-mctl:)ll<,l r-:::Gctions i:l tl..::J G!.br;-·o1ic l1e'1rt
eccv.l:lpr.nying tlk int,r-ov·th . . nd <leve1o ·,r_::mt of tl.lo:- V..,t us
inn0rvction . J . E . Z., 58 .
p·r.- ,;.,:llCJ , ·:r .J . l S32 A t~h.;sioloe> lc cl s tuC.y of th.:: ir1:1erv- tion of tL0
he 2rt of fish er:L.r::os . ?hys . Zool. , 5 .
F:c:;egru. , F . P . 1917 EZ).Jrii...-:entrl stuC.iQs in the ori :dn of th:: vrscu1f'r
andotnGl i u~ end of a r ythrocytas , ~ . . J~ur . An~t ., 21 .
sn~ 0 rar- , E , },~ .
1930 Stuc:!.l...;S ,):1. tn.s Oi.~:bryoloc· of t 1.~ c irculr-tlon in
fisll0s , I. < .. ~.·- II. /'J...... :.r . J .._.~;..tr • .f'..,. n"t ., 46 .
Stoci :-rd. , C . R . 1915 . An 6X)vrir;.entP1 f'n,"'1:rsis of tu;; orlt ln of 'oloocl
<n<l vcsculrr eaC.otl'is1iuc . J,iaL: . \:ist::r I .. ;:::. . Aa~.: t . Bt0L
No. 7 Also A""· • J -.:,ur . Anc:t . 18 .
P . D,
VI
•
E , . .,... ,, ~· 1' I" 1 ·•n .·.Ur'.',..L. _"''OrK'
v_,_.....!...
.i"io.JYV~
.... _
f..le:xanetsr· , 1 . ..1. , 1942 TiL c F· pacity of tje ~Y~ cup of F~nJu1uE tct~ro­
c1utus for- induct i on and rc..gen ~. r>tiou of 1vnses . J. 3 . ·L.. , 91 .
A . . 'ovtBOn , 1,;. R . cr.d .Arr:.s t rong , P . n. l Sc~~ Tl:..s res]irPtory Letrboi.1s:::
of Fundulus het .... r oc litus dur .1.n; e;::~br~ronic C:.evvlopu.vnt .
J . C . C . P . vo l. 2 •
. ."..r· •. strone; , P . b . 1 932 . Th:; c. .. b r ~'onic or~t_in of 1'uact1on in the. '!) r oEephros t h r ou~ .. : c:i f feren t irtion rna. pPr-3nc hy:c,:-v::-sculrr
E'.ssocir.t.i.on . 1'u..0r . J our . !u..r:t . 51,
c::J1cl , C. !,i. l94Z Ox::..a.::tiO.l - roG.uctio!i ~)Pttcr1l i l: d.1VL.:10J.l:~·.l t of r:·
~~l .... ost .
Phys . Zool ., vo1 . 16 .
Cl::)p , C .i,:. lJ98 T:~ -..., rGlr tion of t:.c ox1s of tJ.1~.:: ...::·~bryo to the
first c1c~vr~2 plPn~ . B~o: . Lect . k . L . L .
·c;. ~irt , R . L:: . H~3S R-Q.on<;l ci.cLrEi.i ~--tion in t r1e C.-:;velo_9r.1ent of the
trout . A.~: c.;. . En.t . Ee c h . loS .
F'ile-sov.' , n . 1900 :LntvJicnl.un()si.:lsch::::.lischd Unt..:r~uc:mnu:: :1 rn E ll.>ryon..;n
von J.Ci):::ilSc,r r.; ulG.enstr ·: ..Cll . Arc.~~ . ::::nt . ~ .. Cl! ,, 122 .
- 3-
::1.:lric:1s , l.:. A. &. G~ .. thar , I • .:' . 1~31 Ul- "'violet r ' i~ ion&. ~~1e
prv ction of t\'·i.:;.s "'n<i do utle r.onster s. P.ys. Z -.. 1. val. 4.
fi·J·:::.~lc:' J L.
1S28 0. w~.e locrliL.'='.tion of ~cvc1L_9.t:ent"U~ otencL,e l.n the
e ...brv·o of Fun '..llus ;-.e:tar oc1itus . J . L .Z.., v .... l. b2.
- --- · - ----- 1928 Viscosit:· c:J~nfes C.urin ~ e~rl~ cl,.-· v~ • e st:---:..cs of
Fun~ul~s c~~s.
H~Dn,
Scia nc~ 6S
(~r· 409)
L . . ~ .... 921 l'.1>J •..0t t. )vlic e;r.~Jicnts of V~;; r tebr"'tv e.: ·;.:yos. I.
:'.:1~os t e;r.:br:·o s. B... ol. Bv.ll . 40 .
Kt.e:..n, B . l·~·
1935 Ti1e fe:rt ili zrblt:. _:er iod of the eggs of Fundulus
·. atc.roclltus c:nc sor..;; f' s so ci ~ted p _....;i1.):-.~enf' . B.:..o1 . B tll . 4· •
.K:::. 1 _,c h , Fl' . ... 896 E)...-... ~·~ant-e ll s U .. ~.-..... :r~uchur:_en uab~.... <ian iC;: ...... h::ut r rnu. .:..e r Selr..o:1_de n Vorh . /..."1ft . Cri.s. B~rlin .
----------- 192? o~:;. tlic~.e Vit r lf ;t.r, un .... , bei S\;J}li tf:ernbryonl.3'1
L.:....ltscLr. ftt:r ,j_J<.:;:. f-,.~"1c-t . Forse:•. 101.
1 .. ·, ~!::, \; . H. 190£ 1L~,erE.-..ante1 .1roduc tion of c:·clopi in th0 fish
enbryo . Anet . Rvc. 3 .
------------ 191 2 Ex.,e;ri~GI~ts on locrliz n ti n in t:10 eg of fundulus
het-erocl.:..tus . Anct . R ;c. 6 .
I~tnar , ~ .
1 936 P v ~~~s~r~~unl an rn isoliertan T0~lct~cken 0r . Fv oll enkeL. .• sc.lcib~ .
t,.,).Jl • .t.cz . S'.lJp l·; 9.
--- ------ 1936 Aust c-u sc h von prl'sui~.pt ivar E:. . ider:::is unJ. M~, ' ul f'r:"lrtte
bel'. .. Forellenkain . !....r~:...Er.t. 1Iech. 135.
:~c E·... :.
R.S. 1~30 T.• e eer1·· develo':)r..ent of H~-v ... icur ;)mi s bir;rculrta
\:j t:1 spc:ci.:-1 r af6renc6" to fectors deter.:1i n1.ng th~;;; e:::br y>nic rxis
;/ ..... J.::.' . ~w1ur;:.: • . encl P ;:71·sio1.
vol. 49.
li·..J. _, .:;.r. , T . :a. ! 89 3 EA :~ :r i.:'.:snt-r-1 stud1.e s in tho teleos t eggs . An£1t .
j'_l""~L.. . 8
... --- -- ------ 1 8£:6 Tn-::: fQrr.£. tior. of the fi sh er:.bryo . J. !k:r.;h . 10.
I. J8.:lJ~_C's , J . ::- .
1.~42
R(jt_\.42.. E1 tion e.nd reconstitution in Fundulus.
J·.E. Z . v ol . 90
u ,.._.:_e. ir:..o r) J . 1.:. 1 S 36 e. . The devalo_pr.:ent of i sol r-ted bl £1 stoderr s of
Fundulus he ter o clitus . J. Z .Z. ~ vl. ? 2 •
. ----------·193Gb Tr t.l~ S.:. : l~"l ltr ti on CX~I ;; rh.:.ants on developing t J 2.vvs '.;s .
J,...!.,Z, v~l. 72
---------- 1936c P.~...;cess ~s of loc c.ti z rt:on in d~valopLlg Fu ndu lus.
J . .i;.Z., 'JOl. 73.
---------- 1£38 P·:-:"'c:-:lci~s fur dif::cr~elltiPtion i n tho teleost~cn t>err.1
rin g . J.L. ~ . vol. ?9.
---------- 1939 Thv CP] e city for diffe;rentirt .on of fish e•·:b r'ronic
tissues ir:~p l en t el:. 1.nto c..:.:p'dbi 0n etlbryos . J . .E . L.. , vol. 80.
Pc Gteels, Jecn 1937 Etu des sur les EAst r u l e tion des V .rtabras
l.:...:rob1::- s ti (~UGS. I. Tc2.cos te ~ ns, AJ.·Ch. Biol. vvl. 47.
--------------- 193? :C:t- .... - es sur 1 ~ ge strul r: tion des V; , G3'~-! rGS :r··.0 r obles ti q_ues . I V, Cv .. Cl.lSlOns e~ n~ r ::- l.::s.l.l' C _ . iv cs clo B... o1. vol. 4 8 .
-------------1940 Un r:p e r·c u co::.'lp< r c tif C.e lP t.rstru l r tion chez los
Ch~r4es .
B~~l . R~v. vo l. 15,
Pt .... l.:..)c, F.S.
l'J40 O:~~T ....e:1 cons u:·..!)tion rnli its inhibition i r. t ' a dev 8loy~i0i~.t of FunCi. ' l.l u s & v -..·. - ~ou s ..:elr.gi c fish eggs . Biol. Bull, ?8 .
Solb..:;:rg,
1938 Tu.:. suscr))tibility of Fundulu s het~,..roclitus er:.br.ros
tuX- r . di~tion . J11. "- • .· . ;l. 78.
·
Sp0rL :.
19::.5 Die bi:;;-;·· l r.r..:J Dif te.·,:0 nz i-;:..runc d~s r-:!otoplE's:..rs C.as
~~ L ~Jst - ~ r ·E~ ~s und ihr8 Entsta~un~ . P~oto)lesEr , v o l. 18 •
.S v oc~A.r-d ) J ,F . lGlO
Tb.c o;fe,;cts of "lcoLol < nd )tL.e.r fmestllctl.cs on
or.i.br~{cmic C.:.e vel o·,)r.. ont . J::~ . J :-ur . f.. <' t .
10.
----- -- -1 921 D:..:V'E:lJ)l.~ent r <t::: :1ncl stnuct'J.rrl ex·.; r ~ssion . J .....er .
J . .U • J•. L . t . 28
- 4-
v
~
1.;.3o·
....L s :-_o u ·ez.~r. t ~ .::n.:l.~ ·-... -'-'1 v- .• 'le"' ;-u cours d e
_ ::- ["'S'\;r~ll " ~::,i0n c:.ez S..;:·lli 1 •• Cf'.liC:.ll- .
·.rcl•. :.!.c s.:..ol ., ~? .
~
:.;; :r 0:·. , v.
Utt-:.rsuc!lun,: s: u-g_r ·~.. 1 • .lr.:.e:.)l/"~1 bei.~
-r
R-- .~ ,.(',
.,..,sl· c ......~ t 'e'·
.L"'""'"'r 1......, 1.·1,.,.n,
C: "
"•J. c.' ~'.... c...
...
cl;ntrr ler.. Anlc.gezo .... -:·n . . •.~,- t . ;.l·J ::. , ? c .
·::~11lsy, L.G •
.:i£·33
D ..: velo.'.·:~nt of tie eg:_ of tiL :r;c''SD 1 rt 'i fer 3nt cvr:Gtro:u, te1.~nE.:.rr. tnrt:-; Jcur. Gen. ?h~·s . 16.
\-~J.sae!!bc r g,
V .R.
19:: -';,
·-,
..- .... """L-c
'
n.L .. -,{· · ·· ·•
.. -.. ..
u.:.£.
!'·~s
·- ·...... v
...-
.1 .. ,
•
F· •. uulus ~-:e:~:roc.:!.itus :.:"' b0 rc""'\l:.lr c.~..:>sse \'! it.: Fun·ulus
:.::jc:lis c::u. t .. d •.:";Ckvrol (Scorub.:r sco ..:' rus) . E:;.'1r:·os cr be. €:Otten
fl'.:>!:. r· ci::roc.::>l c:osse~ ~r F . . l~ct~.:r~~l~tus x F . I;~~jrlis Y'llic.> live
· t •• tll £l~ut t~l<:: t1~..8 01 fl"tC.:l.:1[. ,
\,lt~ t."· ~...ec".:.r13l , ;. \',! r.._.r, o:_._y
~r:c conbi.." tlo:l i5 vi--bh: , Fur6.1lus
x Scomt~r
, ""'n · t.:ese :;::Jpy ·e ·;:;lop to c ,v::r..cGd t:: .. br:·oni8 st~g0s.
To c.r·:'-e Fu:1du.lus hetbrocll.t 1 S x F . r:1· jrlis cross ... s, ore sh ld
us.:: (j_,'"' sc:Ld proc odure os · n f ~rt-il i zinJ F . r.~.ct\;rccl it\ s ..;c_ s. Tr-.:..
:.<:c.·:.-::-101 cross :.~u5t ',:>t; ~-.:r:d~ ~t t~1;::; fish tre.. s, ir. rs! .•uc. "'S r.:r-c:-o;;r .... l
C:o :w:. li v ·~ lone c- ft~r b6l!'l. cc.•ut:,ht . f.:;; tlt:: s~ •. '..., tL:10 t trt ::-.ybr1d
e;r ;)SS..:.. s era r;_ "'d0 c o:1 trol c .ll turc;s fr.:>r:: Dot •. pr;r-Jnts s~ :o uld e
str.rtcd . All c ~ ltur~s Lust ba ·rrisc' r-t tjo S"'!'lu t...:r,n ' rrture .
Ii.. stud,:inc.: t:lv 6.-v'"'lo;n.:ant of th0 ,1ybrid.s r tt ... ntion s:1ou:bd
bo paid to cow.pcrr ti ve r F t0s of ut:velopr:l..;n t , e.p:;>G nronco ':l.nd shn;>o
0 :· orgen p1·ir::or<.lic. , I.lorpholot,.y rnC: t pc:s of ]itr:Knts cells, etc.
----------------------R~f er e n c es
Df'ncroft , F. v; . 1 Sl 2 H..:r ... C.it:r of 9i&n~€:ntntion in Fundulus hybrids .
J. E . Z. 1 2 : 15 0 - 1 78 .
GJouri c:h, H. B. 1 935 The develo:!mont of heredi t y color pr'ttorns in
f i sh . A.... . r, ,t . 69 .
F::'.;!:len , H . H. 1 908 Tjt::. .>roc.::ss of he1·ed.ity ~s GXhibitcd by the
devc 1 op!::;.ent of Fundulu s hybrids . J . E . Z.) 5 : 503- 561 .
----------- 1915 D;;,;,·e l o .. )r.:l311t 2nd hc r ~di ty in hct .... ro[enic T8:~Jst
h~rbrid . J. ~ .z
1 8 : v.ll - 576 .
---- - ------ 1 918 ~I~,r br id s be t wet::n Fundulus 2~16. 1>~~ckerel. J .:Z . Z .
Go :.j£1 - 421.
Russel l, Alic e 1 939 P:. :: -er.t inheri trnco 'i ru the FunG. ' lus -Scor.lbL.r .1ybrid
Bi ol. Bull ., 77 : 42 3- 4~ 1.
.. ,
C.'clopi c on d. othar c bnorr:-.ali ties of the i· e['ld ncy be obt: inuC.
Fun,ulus ei:...bryos \·:it: clcohol ,;;['lrly iu evelo~r:1ent
{Stocr:::-r .... '10) . Ey6 abnormclities olon~ cen be obtrined . r trc"'t;:ent
vd tt ~..:gc1 2 •
o·r
tr~"tinc
coas1stently t:,ood rc.sul ts nc.y b0 t:.ottan v:i th dil·ttc s::>lutL:ms
or' r.lcohol in s-.;c v·rt-.;r { :J-9'~) .
60-100 egfs shot.:.lCi b.: pl·~ced in
.fi.r g_8r bo\>'l containing 3~~ ~ lcohol, l'nothvr 60-100 e c.. s in 4/b, etc.
__:, 1ring ecrly cle cv:::ge st~e;os .
Eggs should not be cexposud to this
wrac tman t for L1or.:; then 24 or possibly 36 h.Jurs. Thv_r should ·t.mn
be rer.:oved end plcced in ser \ ·c:ter for ·fUrth-.r develo n.::mt.
Best res_ul ts V!i t~1 l;gCl. 2 ere ot.tf' 1ned by using solutions of 1622 cc. of 11.:. hl[:Cl 2 nede U) to 60 cc. b~· "d· in£ so f' wct.or. Ecgs shou · '.r~
oe plcci.ld ln I.!.::Cl2 svlution shortly eft:r f er tilizr~tion (8 cell stf' ··c )
crd lvft in it for 3 drys, cftsr ~ ich tloy sh~uld be returned to s· e
Vli: ter for further d.::::velopr::ent .
It is advised ti1c:. t results bG r0cord0d in t cbul r fort1 listing
tl10 vnrious ebnornclities obtcinod VJith Gach :'(.0n-v end with G· ell
c_nccntration .
Auel~nnn,
H. B.
1936 .
C.nild , C.1-.l.
1941 .
T.u.::.
:;!TObl0~~
of cyclopic .
( Good bibliogrephy)
~ . R . B . 2 :161-182 , 284-304.
Po.ttorns . nd. Problcns of De velo:1nont .
J?:_) . 270 , 282.
StociG::rd , C. R .
1909 . Th\3 dovE.Jlo:)r..o nt ::;f --:rtific H'1 1.y produced Cyclo_p0:m fish"the Mc.t)nesiu.ru E;::.1bryo ". J. EAp . Zool . 6:285-337.
1910 .
Influ~nce
1-:Lnt .
19 21 .
of e1cohol r nd ~th~r rn~a sthotics on dovalopAm . Jour . Anr' t . 10:369-392 .
Develo:!r:-1o~.1t c l
m12t . 28
retE; c nd.
structur ~:> 1
oxryr0 ssi on .
{Good Bib1iogr~phy)
Px1 . Jo'.lr .
--
• woter
water
'· water
to show the ll'J.mhe:t• of 1iey~ ~ •.:4 b.ou1· D j r <.l'.l ·r t hy !..-!!!!!.1._a ·1e .t::.:;.r-:r.;;.o.;;.;o:;;.;:;.,;:::.::.:~
yos to reach d1tfeJ. ont •3t gef, :· d. ~ve lopm 1 1; l':s.. .n 1ih
1 "'c.. 1le o t te
lation l ) f tho .swim b~adder jt t Lfl.)re hatchin..s 11 tn· V13r ins con11 t1.o
;)f
teraturt3, sali'li ty, ~)to •
1ble
1
.el Me:-r lman, Osbo.~ n Z• olog:loal LL ·)ore ~n · s.nd
·, Yale Uni vera1 ty.
==
..__. .. o;a;~
. . . . . .~--
.,.
..
--
..
~
---
-
1nt;ha:n 0 t;;Cnog:raphlo Labor
If - ~u ;;._·e
; ...... ~
....." ....
r•r·
-~·c..
.. .. ·•
::~c~d
en
'rlO'V 1':1:.::..1:. Rr ~
, · "v ".. '.I...L' t• t.....t:
' e v , .. ~
.. to
·t
it . . "': . . e
. ....L. 1•.:>
...
LJes .
{n
t
..t...
~e~o~s~rrc·o~
M
t::;....
\,i
'!1" t.o__,,. nf th.;.. .... L ~ t SL"J..d
.lSU'"C -.e.'! s ec i::.e!
~ .ic."
s
"' •
···1
... ~, .. -. 4t.J
1
'11-
1..&..
i., .1.U
'"'S)
...,
•
l:'lt: s6'·:t.-s PJ:'3 c.is~i;..t:·u.s! ~ le .
'J:l-l-.. tes,.~.s . .· ti" =>
•· •• ·:,c; tl:cH~ L-- v.e ~J:t.l~ ~t_. t.c yos·.:.eli::
~:t;
s._ 2.l.er -<.:,,.r·.1 ":. .•€' •.• :-l:~s, · .. d ::;.; " sl.ortt..r ' 1
c:o· .oer.
:-1 ::v"" c s ... "'ll )
c.i<et -..i
··.its S~"'l..: L_ t. e c ll·-:1
·er
c:Ft ti.e '1·e . . . Q'•t'- o:~ t.c .,:-ntl~ .
of ..•e fe:."ol0 ere r.:;a j,:.:-i!:~. ~he
se 3 1 thl'OUtJ. t .. f::
1
! •.
ule s ·• ·\. s
'-
:. wlly
' en
~
v
.;.
T .. ·: cccessor:·
re(:;;di~1 <~ s~cs
n
~P. .... l.la.
T-.. S8CJ....-..... e6 ·-: sliM C"("':_ t .. e ...C:!'t:e fr01 th~ .:.Jp:101 t.:> vi'e
th .
(U:...0 l!'1l[L. scu::>scrs1 . '.·.~..;:.~_ force:)s tenr t:! . .-... i~· '·''-·Jl ,;
t.1 :; ovcry L.nd. ':l.lo·,· 'v .. t:: "'t'L s to escr.~e L1:o "dis\1 .n' s2 \'fltJr . r~·
th.:> eggs r.re: f• J.ly :... ~J.YG t .• e_- se.~"r'te ~-..;:<dil} fro-1 t: 0 vn ~~ ftnd
C'V.J8r r be<.:-\Jtif'...<.ll:r vl': J.1S..J"l c lt. < E' e;L. RS .
Ir...~:~tt:r0 t.- t E' ru·v . ;.)t,
tr,->ls:pr.:ro .• r, ~~:1 \.l v.ill no-:; c..: valop .
I.
11orp wlogy of
Stu.ciy Jr:.:.:;ture
ee::~s
t~e
Eg3 .
taken from the; OVf'ry of t: w s C(tid .
1.
'l'h,:; ::.gt.. 1s sur ...·o u:.d0d hy
Lpll]cd to lt .
?..
trans_pE're:1t caorion Plli.cll is clos .... ly
2.
;,t thv ~j_int-:.d G,'d, fll'lu D depression end t:
one h ·· ly -~h rOllt.h it .
Thi.s is tl1e micrO"'J'rle .
~:-.1nutc
c um.. l
-JXtonJ:l.@:
3 . Note tL~ 2olerlty ( blunt Pnd pointed poJ~s) PnJ thu bllrter~lity
of u.c:;:; ,;~,J by tur.JElt. it uvvr .
T~w ~·:.or3 con-i'cn: side of tn .... ,:;f:_( is
t.L futuru " ant5rior " or !..'i.,~clt.l :::icld of t11~ o• .•br~"O .
4.
'I~1i~
l',hc: tJ.1in .2_~2 of cytool · ~ !.1 cov.:;rlnf. t:ltJ ~' olk et thu 9ointc:d
t~1·:.. c..J~c.t-n't oft: ..:-; cytoplrsr:~lc c r·p ty rollinf~ tL0 vf.( .
CJtto~las.,l ~·.illl L5.v"' l'ic_. ·to tnv 0 ..... b:c:ro:r.tic f·t-T:l :JtLll'O~ .
l~ot . . .
poL; .
Stud:
Study a st r iJ:l~ of E:.f:;gs .
TI1.:: e[;., s ~ r.:.. enbeddod in r c~lrtinous
;·..e t :n.x which is produc0d. 'by glr.;r:C.s of tl:~ oviduct , Pnd c o\:L- rod by a
j.:::ll~.' :.:Je:Jbrc:n-.; proJ,w-:..d by tlD nidn":l~n.tnl t:.,l>Jnds .
II
F.S.liT~LIL...ATJvJ ;.~::
1..'T-..:K '-T-::0N
Artiflclr.lly :· . . . r t.i .. l2.ed t..;e,_gs 2:::.,:; i ..o1e frvor cablc fOJ t:1 .j stud::
uf -c:~a first .?h: SClS of d.svolo_)>:•-3li.t t.hcr-. cr<:: those lcid by tho femel0
bvcGasG t~ey l rck the jGl_y envelopes . F-rtilizntion rnl cJ~rv go
C<El bG observed r...:rdil~· if'.. th1s "8y .
HOV.'vV..:.r , tlt0S,; ~f!!-3 . r~;; Vvry
c01"Si tiv..; end ..lUSt 00 .\:v)t ln ·- l1 rt_0 volur:.c of y•rtGr .
QJ.Jt..;n e. ;·;lele. c!.i1d a l'E..r.i.f·l . . sq_uid by noking E l r:mr it:1d.ir.e l
~c.::: tior::. t..a·o .l~)l the .....f'n tl c. ' loac tl1G postori or ( fun 101) s.Ld<:: .
Remove
't£• ..:; l.n!~ s,,c .
In tL..; f...;r...:.-:1" cJ.t tL..:. ovr:r:r o:-cn ""Ll s.u ::"' F~ll trrns-
,.
- c..., -
Pleco ::;~1e :;;s::.cs 1.!. E
solution . 2.li~- t"-.J£'0U@.'-1J..'y .
d.:;pr ..... sP~ . .m s.:'. .:!.C.0
·l· ss
ul.... . .
i'TrD:::il.··r ~. fc\ "'---'ts i
'.r.d vD,,~r.J'l v.1.: ~ r~j_::_l:..n~_-2.!~ u. ckr
Y:
tc!l
"!ld
: s. rr1
·~...1y 1 •• to
:11 L ~ ! ... r:
2 . Al't:..r c1 ...:.crt. rla.•~;, t>: c~t.:))1t.s:-r..ic C9_.) '.51l •·i,.: l'"' f·.'Ol.
chorion, n:r;.6.
c1•:; "~--~~:_.L__:?· ~~l~_inc C'i.)rc~-' i1 · qrpu ~l' .
Thl!'1 is n
1 nC::.ic::n:. Lon o~· f>.lC~&r...sf •lJ. L,-r· .ll l" t1on .
t~ 0
~i-,.s...::::_ f2~"1·_ body_ F;)e'·rs '')ou~. ?,; ~!· .lTtes PI'te ...• ft.:rti.0C .. f;IV8 tl.e 1)_ er ..:::cc•3 yj,' t.l-:, R8C.) (i_ ')01' !_ bvc.j· 1.1d f U.'tL• r
d . . v_si.:>LlJ Gi vLC. )vlLr uoC.Jas (~ -_; ·ioeJley , l'o.~OJ.
o, 1'1__,
1 iz.~1t 1 ..m .
- -----
Plc.ce ::,nc ;_rt:r.ter )f'I·t vf tL!l. fe rt i1 iz0<1 ee:,r::; i.ntr) e l nrce d. ~;lL
.nt.i.1 clc;p·~ see ~·0L-=-r ., ... c' d:) not disturb thd :. for ::,bout 2r-3 l.rs .
T\1t;:· cru set nsi de ::. cr "td!e o·!::s•~rv,1tion of cle . vr ;;e .
fll~8d
III
CL..:; _t V !\GE
I n order to JotaLl a polc.r view of the c·rt.J)1rsi,ic cap Yhicl-t
alolle ·.a:..l unC.\':r·go cler Jat;e it is nbcessr.tr:,.- tu r"o'.l•it t1J.a UfSS ill u:n·ignt
?O~ition .
PL.eG ~- si.. . ell ~ortion of v ose1rne or. e · ry de:;Jrvssion sl1de ,
f..i. 1 .2. it \ll tn :cter und l"lour.t t .. c; e(!,r,s v?ith a h; ir loop so tlPt t:1ay
s-r:,:.ncl. U) . . ..:Ount o ··1 2 ...;gcs i'lhich have both polt.r bou.i';S Ior::JlJu , en(t.
·w tc:l c.: eru. '··t. si10rt i nV: rv uls .
Obc..::rve tlu first ,9l GPVP..L3..§. r>.nd note th..;jr lr'G1etiont!bo the
a:A0s of syn1<.1t::tr; of thi' Gf:/::. • Th:- first c1 er:vr•co:~ 'l l r'ue coHlCldds vrith
tw.. i:'J0dien :.;>lan.:; o:t· t.!...; fut·.u-v c:.loi·yo .
(C o:J.:-.,n1t ~l1 .... fi[s . of \.'r.tf'sC , 1
1 8S1) . The clee.vc~·~ E. is ~~ei'oblc:,s tic, rnd. not s1;1ir<'.J. , in contnet to
other t~ llusc scgs .
IV.
Th0re is
Til.j. T:J P .. w OF
::-·~
du\~ to temp.;rf·ture dii'f'·:rrencus
1-.rE: S 0 r ou~:, cp 1roxi··.1:'tiol'l of th0 tir::.:;s
r~rcheC. .
c oDs::.d_rab1t: vnri:"tion
-: nc<. tlw fo11oy·in b s c:1-::d.ule
V'hlci1 c u:r"Cr:ln
st::[.,~::s
t.r.:;
<.:.
DE~T.GL OPiill~n'
,
-3-
ri __
\J
c. ~+v ..; r
.,1..
.1.~~rt'li7·.t1'on
~-
20 :.;..
G
1 h :
1'
t::s
2r~d
3 ho·rs
;) 1 "'I· body
1st c lc '::..V"' ~~3
12 hou.rs
24 hours
"GestruJ'; ''
= thi..J!~c,
,..
~er~ .•. cr,~l
rlll·"
2 days
3 dc.ys
3~\.
"'
d'1ys
uf' s.11J 11 Clf'lld nnu
aye stPlks.
A~1'~.:-flrDl'.Cu
5~ dcys
SipholPl folds en~ erm~
ey:-s )roj"'ct.
Bi
de.ys
Siphonol folds fusdd into e
Eye st~lks prominent
11
-
Hatching.
12 days
~ppa:r ;
tub~.
v.
SPRbADilJG OF I'HE BLASTJD:i:Rl..
Study eggs about 24 hours aft0r f~rtilization . Lct0r blcstodGrm
stot:,GS v1ill bo sup,lied . No to th'"" r;rsduf 1 cxtGns ion of tho blestodt.rm
about the yclk. The "blBstocone-s u v:~1ic?l f'I'C supposed to gi vo rise to
th-:. yolk cpitll0lius are not very distinct in Loligo . Note. the
thickaning of tlw r:~e.rgin of the blastodsrru ( formction of the .:mtom0 s odcr.m = "gas trul o ti 011 ". )
VI
ORGJN FO!L_.IlTION
It is c onvanient to c 811 tne pol0 whcr e the s;,1ell t lend end nc- ntle appa er "dorsal " nnd tha .,·olk pole "v8ntral ", the siphon s i de "post8rior " end the rttouth sido "cnt-::rior" , nltthough thcsa dosi€,nctions
are not correct from th0 com.porrti va 011etomic c,l point of viGw . Study
a se; quenc·o of at 10 Pst 6 st&[;.a s os rcpr J s-:.nt~d on tl1e ch r rt . Study
the ombryvs from c.ll sides . Mcka drE'v•int_ s of diff0rt:mt st&ges .
Ee.rly s tege s .
( Chart figs . 1 end 2) cf . elso tht3 toxt books of
McBrido and Korschelt~
l: ) sae.ll glc.nd ot dors£11 pole .
2 ) Mantle primordiuru , c::. n e:ctod..;rmel conc<:..ntric fold bonvrth
the shall g~and . ( fig . 2 )
- 4-
3 . r..Ic t.h
--tions
0.:1 tue 1cstarior
-
5. A.Dterl'ii r .. ' )O:.:>t~rior si ~J hor.el folds . Thv L.L'.;lC1 ' . '"
t!1~ prl:J.cl. di.., uf -:_1~; si):.on ;-tht- -Yrt-ter ·:: 11 •'c .. its
c tor .:.~'l:.>c:..-: s.
6.
__
-·8tc~;.)r•
_.... .·sts-
7. Gill
---- ?rl~o~0ie
--·- - - .
8 . The oriGordl£ o:Z' the
~·
9. Note the rhythmic el contrcctions of thz volk e\.itll~J.:l\1]..:..:..
Tney serve.. the yurposa of c1rculet1nl... t:.e llq_u.:lfi _;cl , ell-::
m:=-t-:::riel in the ~·olk src vessels. The r::rt..;;rH·: is crrr1~0
into t he er.brronic tissuas in this '''ey . (Sec ?ort.;lfmn . 1' ''C)
t'iediun. St E't;a s (Chart fie_s. 3 ~nd 4). Noto tl·.e gredurl constn.cti').r
of the yolk sac. The l e. ttt:r continu0s into t i!e er•. br~ro r;hich is thus
forced eround a core of yolk mess.
Obs :.;;r ve:
l) Grovrth of mantle ond of fins.. (The s,_ e 11 elPnd is mannwhile corr-.pletel~r invacin rte; d and not visible).
2) The 0ye
stel~s
e re prominent.
On t ne posterior side:
3) The for Lw tion of the siphon b:· concrasc o nce of tl1e
an tar ior si:p: one.l folds.
Tlle )Osterior siphonol folds
continue e s ridges to the enterior side •.
4)
The
~,
betw.::;en the gill prinordie. •.
Note th;:; further grov:th :)f
yo lk sac, etc.
Old st nte s
ot~10r
prit1ordia; contrections of the
(b0fore hctching: Chrrt figs, 5 and6)
1) The eya st r:l ks E'ra v e ry prominent.. They cont rin the "?ri ::.-.ordia of the optic and ooro orcl go nglia, the so-cAlled
~ihite b odies", Plso c cJntrcl mass of yolk.
2) Tne lens.
T'ne inne- r sec tor v:nich is forr:1ed by the ou tor
~ert of the optic vesicle (~ifferont fro m Ve rt cbrctes ) is
cloqrly visible es e club s l.cped rod extGndinc into the
eye vesicle.
- 5. . . r~o:-~s ,
·.:.:!\: •
. )co
:Jvvel•n.::aL~ o:t
-v:v S.,:. id.
1:-;-;:'<rs .3::>st.ol S .c.
rll
o)
DrJ ·· , G.A. 1 9:.1 S-X'"."'l flcti 'l.ti-:;a f th..; S(1·ti • (L l i o P.:alii)
I. -::op l c.t io, ~;;;l_::_ 1:.-:· il~ r .1d r c r tili.zr+_dl. J. I."or:) .,vo1. 22
··---------· 1 Jl9 II - T.1: s~ " r ~.:!t op~1Cj:e; i s st r u.:-t·.r , n ,H'cu:ctl.on
E'n.: ~o-c_.!"'_::c"1 .
J-='ur. o~ .t:or~:1., •o l. 32.
·f ~:r-c . ~li st .
_,.11 _~c~-, •.
( ;::;_
uv"''-'·r~
\,..;
(G,oJ. Fi['
~-~
.a..~ ·
c:>n ;:.<::.. t. ~-'- UT3.-3
c.p0.·: c~ 0 ..;!""~c:r·•~ . •
!?:! :la-y , L .
r0s 01.
r.,..,..,...... .,.... .,""- 11......Cl-"\.,;, d ''~I "'01 ,_.r..
in ;tZ f'): a-..t:-..:..1~ o-: ... :1-3
.~v\ ·
.t.~
..,
-~'I._
'']
V\;:;~o.
~tr
"', ·C'-.;t
l
.a. 1
_ ·,
;·.. ·-of the
l ·'L·C
l Jc·r~t fo.,...l...<t ... n.l ?tld. ,}2 n~.· 'h1~~ ~ft.'·.: ·olr.r
bo.:.~- :i .:; ~ l :i •) 1l:l j.~r..at:J~t-:.r:..t
s.
..)L"'J .
. •11. ,,c:: , 58
([ _, ·;,, :~~ . ,. '-·~ .n-<'L l l.LS.:.I.UI.f.tl,':} ,
F .... t.'£<.. .? ..... t;···.c~.;.l'L:_ '.3s Dor .....\:tn..,J..s l,:·d ~':!V::t.r:·t·t ... ~:t cL:'"'
.
!~..::-schGlt_.
C, ·).ic.}O )),,c n .
::\:.s~scLrift
f\!fr
Lvuk~\ rt
(G;J~ fl~~ · o: ad vclop:wnt o~ :PtGstind)
- ---·--- ----- l. C•3E "Tf~. r ~l icrh. . ~lCl::l ~nt.\nc: •.l"ngsl ·~ sc 'li(".l t·'J dcr
vol. 2 po. 908·-) CJS. ( Co:J. tr in3 Bi bl iOf.rppLy)
r~ ~.;;J.:.
M· c n:s.ld-.JJ E.·.;, 1914 T-:3xtb.)o:c of 3l·:b r~·ology. I. I,;.v ~rtelJ r [.tn .
r·;-'0f' A. 1928 Dia C.:o;· ~·l ':' .Lopodan, vol. 2; l:;.u..tr::olot:; io. F:-unn u Flo~
d.:;l Golfo C.i NE:_t)oli.
I
(COi.J._~ l0t~ s0rios of fit..urcs of d.ev0l o lr.Dnt of !..u1 i o C''l
plat -- s I- VII; s0rie:. ion of st"' £,8 8. Consult _:li"TT,icu~. · 1l.\'
pl c tG 7, :tit..,S . 4 rnd. 4a for na·v,.ly h :'tc :. .:; sc;u i.C. . Nvt6 t 1. ·1
N<-<:1' dvscribes tfi.:; l~I~~G. it0rrcnvcn s:p"' ci c:.s , L. v . . . lt:_,ris , ':!.l·c;;
d..:v..slo_ls mo:...:: slowly then L. P. )
Portmcnn, A. lq26 D.)r e;..lbr:: onc l e KJ:-......... ::1::-.ui' unrt div Dott~rr-.Jsor:;tlOlt
boi L.:-.liLO· Zuitscllrif't f. 1\~vr~•hol. u. ffl{ol. vol . 5
( 0 ood fi:s. of JOlk s Fc circul ~ ti on).
?vr tr:l8nn, A. end Biider l £28 Y:)lk a 'csor.}tion in Lvli ...o nnd t '10 function of the amtr~·oni c li vcr. ~u e rt. Jour. l\I.L.Cl'O. Sci.
vol. 72.
R"nzi, s. 19.31 Svllu;:)o <li ;>F.rti isolate di oubrioni di C(,:f'~lo1"odi
Pubbl. C.cll c S :.::.zione Zool. di N:._Joli, Vol. II.
---------
1931
vol.
Duplicit as cruci c t e in e1·:r.brioni di C0f(' lododi. ibid.
II.
1934 Ueber Lo.l.e bi)Ol c re Diff-....renziarung clos Cv~hrlo :-reden -und
Prosobranchiar-Eies. Roux. Archiv. v ol. 131.
\":1 tast: , S.
1991 Studi v5 on Ct~,l)lle.lopods. I. Cl eev ac;e , J. of Iv;or~h.
vol. 4 (S~e fisures)
v:illieUJ.s, L.V;,
The R:latomy of the Corr.:..10n Squid. {Ex cellent fi~,..ures
of the structur e s of tne ~du lt squid.)
Spe;lc , J.
CRUSTAC~
Libinia, (The Spider Crab)
Examine fenale crabs which are carr,ying eggs. The color of the eggs gives
sooe Jndication of their stage of development, the youngest eggs being bright vernill~on while older stages are bright red.
The oldest chocolate colored eggs are
not suitable for study.
With forceps renovo a few hundred eggs (each student should take then from
his own specimen) and place thee in a finger bowl from which tho rest of the class
oey take saoples.
The earliest stages of division cannot be seen in tho living ogg. Protoplasmic division occurs after the fourth nuclear division at uhich tioo the nuclei
have migrated to the periphery. Find several stages of cloavage. Are the nuclei
visible?
Later stages of development can best bo seen by reflected light under low
power after the eggs have been placed in strong HgC12 which whitens the embryonic
area. Find embryos which show:
c)
d)
the 5 embryonic rudiments (examine by roflectod light)
the nauplius stago with the first 3 pairs of appendages.
Are any biramous at this tine?
later stages hnving 5 or more pairs of appendages. In these the
stomodauo, ventrnl fold, dorsal shield, telson, and ganglia
may be found.
tho Zoaea stage (from cultures prepared by the instructor). Quiet
the larvae with lAgS04 or other a.no.esthetic. Under tho high
power, details of muscles, tho coopound eyes, the contrRctile
honrt and intestine can be observed.
If Hippa females are available, remove and exnmino eggs as for Libinia.
Cleavage stages are more frequently found for this form.
Balanus eburnous (The Barnacle)
With u strong knife break open the shell and find the egg lnmelln at the base.
If the eggs are ripe and in the process of development, the lamella is firm and
can be taken out entire. Place it in a watchglass of sen ~ator and tense out a
few eg~ s v1i th a needle.
Fertilization is internal. In the youngest stages nvnilnblo you can observe
the polar bodies, and the approach of the germ nuclei. This is best soon in preparations that have dried a little so that tho eggs nre slie-htly prossed by the
cover glass.
Study the cleaving egg up to the 16-cell stnge. These eggs are delicate;
usually the sace lot cannot be \vtltched through all of those cleavages. tf they
die, find other egr,s in the stage that you need.
Later stages of interest are: a) gastrula; b) tho 3 aegcont stage; c)
c) the 5 segment stage; d) the nauplius just hatched; e) tho nRuplius after
tho first and second moult.
In tho nauplius notice tho appendages. tho labrum. the median eye, paired
Ganglia or brain, the digestive tract.
- 1-
If goose neck barnacles are available, remove the ~ egg lamella from the
body by breaking open the shell, and observe as for ~~lanus . During development
the blue pigment in the eggs changes thru violet to pink in the nauplii.
Literature List on Crustacea
Bigelo~. 1902, Early development of Lopns. Bull. !!us. Comp. Zool. Harvard, 4o.
Binford, 1913, Fertilization in Monippe. J. IJorph. 24.
Brooks, Embryology and Metamorphosis of Uacrourn.
Dnvydoff, Truite d 1 Embryologio comparoo des Invertebres (with good literature list.)
Croom, 1894, Early development of Cirripedia. Phil. Trans. Roy. Soc. Lond. 1~? B,ll9.
Herz, L. E., 1933. rho Morphology of the lntor stages of Balanus cronntus.
ol.
Bull., 64, 432.
Korschclt and~cider, 1899. Textbook of Embryology II.
HacBride , 1914 Textbook of Embryo-logy.
guller, F., 1864, Ffir Darwin.
Richards, Aute, 1931. Outline of Comparative Embryology, p. 162 nnd Chnpter !Il
(rccnpitulntion theory). John Wiley, New York.
Thompson, _J. v. 1835. Discovery of the Metamorphosis in the 2nd type of tho Cirripedes, ate. Phil. Trans. Roy. Soc. London, II, p. 385.
Artomia (crustacean)
Th..; "brine shriop 11 ArtomiP. is o. primitive (Phyllopod) crustnccun which is
found throughout those po.rts of the world which ero sufficiently arid to havo lakes
and other bodies of wator which are more saline than tho sea. The adults are about
half an inch long and are very abundnnt in such saline bodies us tho Great Snlt
Lake (Utah) (7 to 11 times ns sa.lty as the sea), Mono Lnko (Calif.), etc. They
nlso occur in abundance in salterns in which sen water is evaporated in the sun to
manufacture snlt, as on the margins of Snn Francisco BQY, uhore the cysts used in
the Laboratory wore obtained.
Artemia dsvelop and live well in sen wntor, but they nrc rarely found in the
sen in nature, probably because they are defenseless and are eaten. Their tolerance for salt is so great that they can thrive in strong brines in which they have
no enemies .
The cysts, or 11 eggs" as they are sometimes called, are produced in groat nunbars at certain seasons and float on the brine so that they may be blown in windrows on tho shore by wind. These cysts are actually nauplii in a dried and resting
condition within the original egg shell.
They keep for years. Whan plo.ced in a suitable medium, such as soa water,
they absorb >70.ter and hn.tch in 20-45 hours, depending on temperatura, etc.
Exn.~ine somo of tho dried cysts under the nicroscope.
They are ~rk brown
or sepia colored and do not transoit light. When dr,y, they nro tho shape of n
sphere which has boon caved in on ono side. After soaking in sen water for an
hour or two they absorb enough water to round out .
Examine some enbryos in the process of hatching. Tho hard, outer shell crncks,
ar.d the ~~uplius energos in a quiescent state within a delicate membrane or sac which
renains a.tte.ched to the shell for some hours. Tho nauplius begins to move and finally breaks out of tho sue to swim about. Examine so~c recently hatched nauplii.
They arc so active that it is nocessnry to o.nesthetizo them, and they aro so hardy
(the adults live o.nd swim for some time in Bouin's fluid) . that it is difficult to
a~csthotize them, but they will finally come to rest in strong alcohol.
Observe
tho 3 pairs of cephalic appendages , the single median eye , and on tho ventral side
tho enormous labrum or upper lip. The digestive tract may also be soon.
- 2-
Observe sooe larvae ~vhich h..".Vi... oc.1 !~~tchod for 2 or 3 days. Evon if tho
lnrvne are not fed, tho first ooult t~~ c s 1l~~c to foro o notnnnuplius. This differs froo the recc!'ltly hatched nnupli ~ • 1' 1:\C;.. nlly in oing longer nnd larger nnd
in having the buds of 3 ooro puirs of s ·~ ·none; ~p endnt;os (on tho thoro.x). No to the
stronG positive phototropisn of the stock culture.
If the older stages arc available, observe then also. Adulthood is reached
after 14 or 15 moults (Heath) during which the number of appendages gradually incrC(l.ses grant ly.
Bibliogrnphy for Artenia
1.
2.
3·
4.
5.
6.
7.
Bond, Richard M•• 1933. A contribution to the study of the n~tur~l food cycle
in aquatic environnents. Bull. Bi~han Ocean. Coll., 4, article 4.
Boone, Eleanor nnd L. G. 14~'\s-Bo ing, 1931. Sii!t effects on eggs l".nd
nauplii of Artonin salina L.J. Gen. Phyeiol., 14, 753.
Fransoooir, Luise, !939. mrngc der HorkUnftdes Uetannuplialcn Mesodorns
und dio Segnentbildung boi Artemia salina Leach. z. wiss. Zool., g52: 439-472.
Heath, Hnrold, 1924. Tho external development of certain Phyllopo s, J. l4orph.,
38~ lli·
tOchhead, J. H., 1939. Molting and fo~.tion of tho exoskeleton in tho brine
shriop, Artooia gracilis Verrill. Anat. Roc., 75: 108.
LochheA.d, J. H. and Margaret S. Lochhead, l94o.
egg shells of the brine
shri~, Artcoia. Anat. Roc.,~~ 75-76.
VThi taker, I>. 1.1. 194o. Tho toloro.nco of Artemia cysts for cold and high vncuum.
J. Exp. Zool. ~· 391.
--n.e
TUN I CATES
1944
A comprehensive survey of developoental processes in the tunicates would include exaoinQtion not only of oges 8nd embryos but ~lso of ~etnmorphosing larvae,
vcrious types of vegetative reproduction, and regenerating forms. Because these
developmental foros must be constantly related to the structure of larvae and adults,
it is neces~ry that the student should first of all review in a standard text tho
anatony and life history of the common types of tunicates.
A.
SIMPLE ASCIDIAUS
DEvELOPMENT TO THE TADPOLE STAGE.
Styela.
(Old name Cynthia)
Conklin 1 s classic description of the development of Styola eggs (1905) provides the morphological bo.ckground for the oxperi~onts (Conklin 1 31, ~ 1 34,
Dalcq 1 38, Rose '39• Tung, Ku & Tung 1 41. etc.) which are actively shaping our conception of the organization of Tunicate eggs and enbryos. Conklin's figures should
certainly be referred to during the following studios.
a..
Mathods
Though truly hermaphroditic, Styoln is ordinarily self~sterile like several
other Ascidinns (Horgan 1 38). It sheds tho eggs and sperm between 4 and 7 p.m.,
and fertilization takes place when ripe gametes froo two different individuals cot
mixed. It is easy to have eggs shod and fertilized on nor~l schedule in tho l~b­
oratory, but this entails the disadvantage of having to study tho migrations of
yellow pigment ~ithin them by artificial light.
The classic method of obtaining oggs and embryos from Styela has been to
mince the goneds from a number of large individuals together in a dish of sea water"
- 3-
This liberates all stages in the r.~turation of eggs and sperm, and usually a few
eggs will be fertilized, whatever the time of day or night, ~nd will commence norcal development.
s. IJ. Rose '39 has developed a method of controlling the natural spawning in
the laboratory by illumination, and this is the best way to get fertilization for
experimental material . lt works W·J ll oxcopt for o. fow ~Teaks in mid-sumner, whon
the animals are spent. By such control, tho same batch of aninnls can be induced
to shed a number of times on succossiva dcys.. They are kept darkened until eleven
or t~olve hours before fertilization is desired, und thon nn artificial day is
started by turning on en electric lig~t. A 4o \'lntt bulb 18 inches from the o.nimuls
is sufficient. Eggs and sporo ore discharged in clouds at the dosirod time.
b.
V.ature Unfertilized
Egg.
Sketch the mture unfertilized oge, Phich should show the following:
Chorion, n tough monbrnnc ,.,i th perhaps a fov: follicle colls ndhering
to its outer surface.
Small spherical inner follicle cells ( 11 Nurso colls 11 ) between chorion
and egg itself. They contain yellow granules.
Peripheral layer of egg, n clenr layer containing minute yellow
granules .
Central part , consisting of gray yolk platelets.
Germinal vesicle, a very large eccentricr~ly placed nnss .
c.
Post-Fertilization Rearrangenents.
This is a difficult process to observe es nost eggs havo too little pignent,
and feu are fertile in 11 mincod 11 cultures. It is better to onit th~s section and
to concentrate on clenvage .and gastrulation. Then, if tine is available and if
tho eggs this year contain sufficient pigment, these renrrnngenents nay bo studied.
Carefully watch for the roarrangonent of egg substances starting within 2-8
oinutes nfter fertilization. Tho cloo.r yellowish peripheral nattor strenns to tho
lower pole over tho yolk, follo~ed by the clear protoplasn from the animal pole .
If the yello~• pigment is not visible, try a ooro brightly colored ogg . Use only
daylight for illumination, and have the diaphr~~m on the microscope wide open.
Roll the egg around by ooving the cover slip with a noodle. Tho yellow inner
follicle cells may nlso be migrating; watch tho egg cortex.
Tho gray yolk rises to occupy the upper pole, all except the space that surrounds the maturation-spindle complex~ Soon tho yellow substance nccunulates on
one part of tho lower hemisphere, where it assumes crescentic form. Immediately
above the broad part of the yellow crescent is a layer fornod by the clear cytoplasm.
Tho most remarkable characteristic of Styeln is that now at tho conclusion
of those movements the position of tho future lnrvo. is visibly marked on the surface of the uncleo.vod egg. The broadest part of tho yellow crescent is at the
posterior pole, and the horns wrap nround the right nnd left sides. The nninol
pole, where the germinal vesicle lay, becomes the ventral-anterior side of tho
lnrva. The vegetal pole, where tho spermatozoon entered. , is tho future dorsal
side. To avoid confusion, fix firmly in your nind the relationship bct~oon eg~
orientation and definitive axis of cobrJo.
d.
Cleavage
The following approximate tine schedule for tho embryoloGY of Styeln is from
tho great oonograph of Conklin ( '05a). Follow the events of clo1wage in ns 1:1uch
detail as possible . Observe gnstrulation and watch tho tadpole take shape. Skotcn
-4-
a succession of cleavage stages, showing bilateral synoetry and location of yellow
crescent oaterial. Sketch at least 2 stages of gastrulation and 2 of elongating prot d ales
(This schedule is for normally shed eggs. If eggs are obtained from
''~in~edll ~ultures, cleavage is dela..ved, the eggs apparently maturing at variable intervals after striking the sea water.)
First cleavage to 2 cells after 4o minutes .
II
II
II
II
II
4
30
2nd
II
II
II
II
II
8
30
~rd
II
II
II
II 16
20
th
II
II
n
II
II
20
5th
II
II
tl ~~
II
20
6th
II
II
II
II
11 112
20
7th
II
II
n
11 218
20
8th
To neural plate stage, "2 oore hours.
Fully foroed tadpole 12 hours after fertilization.
"
"
(1)
First cleavage. Equal, separating the two horns of the yellow
crescent froo each other, likewise bisecting the clear protoplasm
anterior to tho yellow.
{2)
Second cleavage. Nearly equal, vorticnl, at right nngles to tho
first. The two posterior cells (E3 on the left and B3 on the
right) contain little yolk and practically all the yellow eroscent substance. Tho two anterior cells (A3 on the left and ~
on the right) contain much yolk, and practically nono of the yellow crescent substance. Tho clear protoplasm goes equally to the
four cells.
Third cleavage. Horizontal, the upper quadrant (cells a4, a4, b4,
and b4) sooewhat smaller than the lower quadrant(cells A4, !4, B4
and~.
Tho yellow crescent substance is alnoGt entirely confined
to tne t'rlo posterior dorsal. cells (B4, E4).
( 4)
Fourth cleavage. The planes of cleavage vary in different quadrants, but the new cells do not overlap the sagittal plane of tho
eobryo. Two of the antero-dorsal cells and two of the posteroventral cells of the 16-cell onbryo are crowded awqy fro~ this
sagittal plane, but all other cells touch it. The dorsal and
ventral hemispheres at this stage are mirror images. The yellow
pigment lies in four posterior cells (E5.1, B5.2, E5.1 and E5.2).
Fifth cleavage. Cleavcge in the dorsal (vegetal) hemisphere precedes that in the ventral (nnimal) hemisphere, and cleavage in the
anterior part of each hemisphere precedes that in the posterior
part ..
At the 32-cell stage, the yellow substance is almost entirely confined to six dorsa-posterior cells, three on eaCh side of the
midline (E6.2, E6.3, B6.4, E6,2, E6.3, B6.4). They give rise to
mesoderm and ~esenchyme.
Six yoLl<:-filled cells at tho vogetal (definitivofurso-posterior)
pole anterior to the yellow mesoderm cells, will give rise to
endoderr.1: A6.I,. A6.3, E6.1, A6.1, A6.3, B6.1.
Four cells at th~ anterior border of tho embryo just below the
equator (A6.2, A6.4, A6.2 and A6.4), nnd two just above tho equator (a6.5 and a6.5) wrrr-give rise to tho notocord and neural plato.
All the rest of the cells are ectodermal.
- 5-
(6
Later cleavage, gastrulation, neurulation. The gastrula passes
through disc-shaped, saucer-shaped and cup-shaped stages, starting with the 7th cleavage. As it finally bacooos egg-shaped,
the gastrula's blastopore, found at the sna.ll hind end, becones
~shaped, the stem of the T bordered by the yellow cesodermoasenchyno cells.
The cells overhanging the crossbar of tho T-shaped bl<stoporo
constitute its dorsal lip. They ovorgroq it, finally engulfing
the yellow cells which froo then on are only seen dimly through
the translucent ectoderm,
Watch the gradual elongation of the embryo and tho appearance of
definite tadpole form.
~olgule..
The cges of Molgula follow almost exactly the pattern of development s en in
Styela, but they do not have pigment. The animals are self-fertile. A few fertilized eggs c1o.y be obte.inod at any tioe by mincinr, tho gonnds in sea wnt er, although
spawnin norl:'.n.lly occurs at daybreruc.
Rose's method of controlling the spawning of Molgula or Cionn is to put a.
few individuals in a largo dish of sea wo.tor and store them in the dnrlc, for instanre
in r. desk drc.wor, until egr,s arc needed. Molguln '17ill shed 15 minutes or so nftor
boine; brought out into the light, Ciona ir:unodiately.
Stnrt sone Molgula eggs developing, and follo'l7 the cleavnge through gastrulation. If developing eggs nre isolntod in a watch glnss, tadpoles and young st~es
of metamorphosis nre obtainable. Tadpoles arc fully forced in 8 hours. (Conklin
t 05a)
(Grave 1 26).
B.
1.
AMA.ROUCIUM:
COLONIAL ASCIDIANS
TADPOLES, l.1ETAMORPHOSIS , EPICARDIAL BUDDI NG.
(Usually not r.vailable until July)
a)
l-1ethods.
Tha larvne of Styoln and lAolgula are so small that study of their intcrnv.l
orgnnize.tion is difficult . The compound nscidian Arnaroucium is viviparous, and the
large tndpolcs are easily obtained fron pnrent colonies, provided the nnterial is
ripe and has been quite recently collected.
Squeeze a ma~s of fresh Amarouciurn over a little sea '17ator in a fingerbowl.
M['.ny highly colored frngncnts of the adult individuals will be ejected, together
with eggs and embryos. Fill tho bowl with water nnd pour off the coarser pa.rticles
whirling ncar the top . Tadpoles and eggs in all stages of development may presently
be found at the bottom.
Tadpoles usually leave tho parent colony shortly after sunrise. These undamaged P~d fully developed individuals can be collected in the laboratory, and they
arc tho best naterial for the study of swimming and of metamorphosis. They collect
at tho top of the vater on tho side of the tank nearest tho lit;ht .
If colonies are kept in a shrouded aquarium tho shedding co.n be postponed
a more seasonable hour. Swarns of active tadpoles usually npponr within fifteen minutes after bringing ripe colonies out of the dark. About ~ third of th
tadpoles will emerge within half an hour, if illuminnted first at nino o'clock in
the morning; if tho colony is kept dark until mid-afternoon about throe-quarter&
of theo uill emerge within half nn hour. Nearly all of them will commence metamo~ ·
phosis within an hour of being shed.
~~til
- 6-
b.
Early stages.
Fro~ the debris in the f ingerbonl, sdect and sketch a series of embryos and
eveloning larvae that shm1 the gradual energence of tadpole forn. None of these
squeezed tadpoles \'lill s•1io innediately, and few 11ill attach noroally or develop
further. ~witching oovenents like those of vertebrate enbryos or fetuses will be
observed.
c.
Tadpole structure and Behavior.
Watch ~rouciuo tadpoles swarning in a dish of s en water. What are the rea cti ons to light and gravity? How do they propel themselves? Place a few tadpoles
on a slide in a snail drop of water and study their anatony un er th nicroscope .
(Grave 1 20, 1 21.)
Not all structures can be seen in a living tadpole. Noto the thick test with
contained test-cells, the adhesive papillae, the atrial siphon (near the tnil), the
oral siphon (usually l a rger),; and tho sensory vesicle b etween tho two siphons. Tho
5 ~,11 pigment cup with lens is a light-perceiving organ, ~nd th ero is a snnll round
black st~tic orgnn also within the sensory vesicle. In tho tnil, note the muscl e
cells ~nd notocord.
Sone Amarouciuo tadpoles have been fixed in Bouins and stainvd with bornx
carmine. In one of these stnined tadpol es , identify tho pharynx ,.:i th its visceral
clefts, tho centrnl yolk mass, the intestines and the nervous system. (Grave 1 21 ).
Sketch e tadpole, showing structural details seen,
d.
l4etamoryhosis.
Isolate a dozen or so tadpoles in sepnruto drops of wat er in watch glnsses,
for study of behaviour during metamorphosis. When in the course of the next hour
the tadpoles have firmly attached to the glass, add moro sea water to tho dish . After attachnent (sometimes before there is time to ~ ttach, the flesh of the tnil is
wither ed nnd drawn in, tho test s11ells and metanorphosis has started. Within n couple of hours, movencnts of tho body mny be obs erved. Th e results of the extensive
and rapid intornnl reorganization that is going on can be observ ed b est two days or
a week later. (Grave 1 35). Make several timed sketches of the ext ernnl aspects of
metamorphos is.
c,
Later Stages.
Observe and sketch netamorphosed Amn.roucium individuals i7hich have be en
f or four days or so after nttachment. They are fastened to watch glasses
which have been stored in frames under water. Gently flush debris from them nt th e
t?.p , and avoid tipping off their cover of water, After l!lf'Jdng your records you ·;fill
r eturn the specimens, still living in their watch glasses , to the fre.mos they cnmc
from.
gro~ing
Examine the specimens under low p0\7Gr first, for orientation . If they a re
upright they may be flnttoned out by gently lowering a cover glnss on them.
l If groivn on cover slips, turn it over) •
~rowing
At one end of this animal are the ntrinl nnd ornl siphons, at the o th er is
the po st-abdomen, with tho heart at its tip. Wntch the hcnrt for periodic reversal
of beRt. Identify the epicardium , c usua lly pigmented strand of tissue running
throughout the postabdomen from pharynx to heart reg ion. It is nn ngent in nsexual
r eproducti on and colony formation.
Below tho siphons is the ph~ rynx with its thr oe rows of numerous stigna t n
( viscernl clefts). It opens into a short esophagus \7hich conn ect s \7i th n ro,l: d .•rc~ ­
l ow stot:~ach r.1arkod by muscular bands. Tho int es tine turns sharply nfter l c::-.v". :1•; tJ. £>
stomnch, and ends ncar the atrial siphon. The endostyle is sharply indicated or.
- 7-
the r,all of the pharynx, delimiting the two atrial pouches.
the pharynx, and open to the exterior at the atrial siphon.
f.
~icardial
These pouches surround
Budding.
Asexual reproduction of ne7 individuals may be seen in laboratory cultures
about 14 or 15 days ~fter attachment of the tadpole.
r, swarms of buds in all
stages of gronth and migration cnn usually be found at the bases f the tiniest transparent fingerlike lobes of a large healthy colony . Demonstrations will be m~de of
them, which should be sketched at low me~nification.
Asexual reproduction is accomplished by strobilization, i.e. segmentation of
the post-abdomen \7hich contains the epicardial strand. The buds consist at first of
inner vesicle (from epi~dium) r~d outer covering (from parent epidermis). All internal organs of the nmv individunls foro fron the epicnrdium tissue, which nEls n
phnryngea.l derivative, i.e. endoderm .
This method of asoxunl reproduction is distinguished from others in Tunicates
by being called Pharyngeal or Epicardial Budding, (Kownlevsky 1 74, Berrill '35).
The epicardial buds while differentiating into new zooids movo up and truce their
place around the ~~rent . During the strobilization of the ~~rent' postnbdomon, tho
old heart is isolated and dcgcnoratos, and ~ new honrt is regenerated in tho parent .
2-
BOTRYLIUS:
TADPOLES, UETAMORPHOSISl J.TRIAL BUDDING .
Botryllus is enother compound Ascidi~n, which is found encrusting on ~ocks,
wharves o.nd floats. Certc.in colonies, brought into tho laboratory and placed in
dishes of sea ~tor, will liberate ~ny tadpoles. Tadpoles and nll stages of cleavage and development ony nlso be obtninod by nincing tho colonies and hunting in the
debris. If the normally shod tadpoles are placed in a l i ttlo son wrtor in n r:ntch
glass, they soon nttnch to the dish and conmenco their rapid metamorphosis . (Herdman
F, C. '24).
a)
Structure of Tadpole.
The tadpole is not as large as that of Amnroucium, but shows nn interesting
new feature . Just under the adhesive papillae is a ring of 8 ampullae which aro
diverticula of the body wall, destined to be parts of the as yet non-functionnl and
incomplete circulatory system (Grave and Woodbridge 1 24). In metamorphosis they
unfold like petals of a flower, and spread around the edge of the developing tunicate. As tho colony grous these increase in number.
Xn addition, identify the statolith, a densely black cup suspended in the
sensory vesicle by a slender stalk and closely associated with light-sonsative elements (Grave and Riley '35); the t wo siphons; tho pharynx with several stigmAta;
the stomach and intestine. Sketch tho tadpole.
Tadpoles attach to a substrate within an hour or two after hatching. Tho
metamorphosis is extremely rap id. (Grave '35· Grave and Nichol 1 39). They often
netnmorphose without attaching and stick doun a day or so later.
b,
Two-Day-Old form.
Atrial Budding.
Tadpoles have boon allowed to nttcch to watch glasses nnd grow for two days.
They are now larger and clearer, and usually so oriented that the observer looks
directly down into the atrio.l and oral siphons . The lnrgo pharynx is in tho shape
of a truncated cone and bears up to three rows of stigmo.:~n (visceral clefts) Vlhich
let Hnter pass out into tho atrial cavi tios of the two siC.os. Tho endostyle lies on
tho under side of the pharynx and thus appears ~s a. rod under the mouth.
The stooach ordinarily appears as a yell0\7 body under the atrial opening .
The intestine, after turning to one side fror.1 tho s ton.:'lch, returns to empty nonr
tho ntriun. The pulsating heart nnd its vessels o.re of particular intorost,
- 8-
espocinlly tho flow of blood nround tho oouth
~nd pharynx.
A now bud (First blnstozooid) appears from tho sexually devel ped nnimnl (the
oozoid) ns nn ovnginnticn of tho atrium at one side. It is furnished qith a blood
tupply, and presently tho diffcrentinting parts cnn be distin&Uishon. Coming from
\he atrium, the nholc bud, end all parts of tho no~ in ividual, nre derived from
octodern. This is the Atrinl type of budding. (Borrill 1 41). Sketch an oozooid
showing buds.
c.
Week-Old form.
In these animals, general structure already studied may be easily seen under
power. The pharynx of the oozoid has developed 4 rows of stigmata, and the
first blastozooid may also have three or four rows. Blastozooid buds of the second
and third order oa.y be present.
lo~
The first blastozooid bud in Botryllus is single, all the later ones are in
symmetrical pairs. The same organ structures may be seen in all the individuals,
notwithstanding their diverse embryology, with the minor exception that the oozooid
does not develop gonads. Through rearrangement of individuals, the completed colony
shou~ a comnon atrial pit in tho center, with separate oral openings nt the periphery.
For the anatomy of the zooids and structure of various stages in bud development co~are the living preparation in tho watch glasses with fixed-stainod-mounted
preparations thnt aro uvc.ilablo. These \7oro made from n colony that wo.s nearly
transparent whon living. Most colonies develop so ouch pigment in tho tunic that
~hole mounts aro useless.
Ench bud consists at first of a disc, then n sphere.
The sphere extrudes sox cells at one or both sides and becomes partitioned into
three vesicles, tho lateral ones foroing nt real chnobers, the middle one tho
pharynx. h~tor stages show differentiation of the rest of the organs fro~ tho
pharynx-vesicle. (Borrill, 1 41).
3.
PEROPHORA:
SEPTAL BUDDING.
Perophora is a little green-colored ascidian, which by oeans of stolons forms
loose colonies on ;y-harf pilings, etc. Pieces of the colony mv.y be gently stuck to
watch glnsses ~ith vaseline, and stored in running sea water. After a d~y or so,
stolons ~ill be sent out over the surface of the glass and non blnstozooids ~ill be
formed ut intervals. E~nmine a watch-glass culture of Porophora ~hich hns been
growing for tno weeks,
Notice the brunching pattern of the stolons. Tho tips show explora tory tendencies like s~ll pseudopodia. Tho outgoing and incoming blood strenms in the
stolons are separated by a mesenchyme septum. All stagDs in the formation of new
individuals will be found, nrranged like pumpkins on a vine, nith tho youngest nearest tho tips. The youngest buds consist of an outer vesicle derived from the epidero of the colony ~nd an inner vesicle formed by the splitting of the mesenchymatous stolon septum.
Organogenesis takes plnce in the innor vesicle, which is derived fron mesenchyme (Bcrrill '35). This method of buddinb is distinguished as the septnl type.
EMBRYOLOGY OF IJOLLUSCA
Gastropoda
1.
The Voliger Larva.
Study the typical Veliger larva of Crepiduln fornicota. Obtain material by
breaking the ~imal from tho substrate to which it is attnchod. You will find the
yellowish eggs {ondosod in transparent capsules} attached to tho substrate or in
- 9-
the sheil of the ::~other. Tease the eobryos out of the capsuitht; Obtain swimming
larvae in different stages, particularly old ones 1 ith the yolk re&orbed, They
are transparent and show the inner organs. Consult Conklin (•97) figs, 80-82 Rnd
the textbooks of Parker-Haswell vol . 1 (1928) fig. 657 on p. 734, or MacBride figs.
263 ff. on pages 361 ff, or Korschelt, 1936, Yol. 2, pp. 881-891. StudY dorsal,
ventral and lateral views. Qouth and foot are on tho ventral side, the anus is
on the right side.
_•ote:
1.
2.
a:
5.
6.
7.
8.
9.
10.
11.
Velum with powerful cilia.
Head vesicle (dorsal)
Eyes with lens (dorsal)
Foot (ventral), study it in lateral view.
Statocysts (ventral, on basis of foot).
Mouth with J?Owerful cilia (ventral, above the foot.
Oesophagus (dorsal), stomach, liver. The different parts of
the intestina onn be distinguished only in older embryos
which have resorbed the yolk.
Anus (on right sldo).
External kidneys (lateral to foot.)
Heart (dorsal) •. Observe tho heart beat in older enbryos.
Transparent shell.
II.
EARLY STAGES OF THE VELIGER LARVA
There is no typical trochophors stage in Crepiduln. Study s~os of direct
transformation of the gastrula into tho Velieer larvn. (Conklin, '92, figs . 77-79).
Observe the grudunl development of sholl gland, shell, velum and foot.
Literntu:ro;
Conklin, E. G., 1897. Embryology of Crepidula. J. Morpho!. vol'. 13.
Morih, c. E. • 1938, Tho Anatomy of the Gastropod, Crepidula .adunca.
Univ, Calif. Publ. Zool., vol. 43, 83-92.
1939. Organogenesis in the Gnstropod Cropiduln adunca. Ibid. vol. 43, 217-248 .
La.melli'branchia (:r:'olec:rpodn)
III.
The Voliger Stage.
Study the Volger lnrvn of Cuningia. Study larvae 2, 3• 5, and 8 days old;
mount them with a few shreds of lens paper to entangle then or use Chlornlhydr~to
as an anesthetic. It will be necessary to find more or less quiet individuals
~ith veluo extended.
Contracted individuals will not do.
Those •nhich are expanding and contracting the voluo are best for study
since this moves the digestive tract, Use tho low power first nnd after a good
specimen is found turn on the hieh power. ll.'1.ko out tho following:
1.
shell.
The general shape of the body with straight hinge line and semicircular
2.
The ciliated velum and apical tuft,
The digestive tract consisting of oesoph~s, stomach and intestine.
Is the latter coiled or straight?
4. Position of mouth with reference to the velum. Also position of anus.
5. Ciliation of the digestive tract. How much of it is ciliated? What is
the mode of feeding? Add Chinese ink to the water ,
6. The primary body cavity is a rather large space seen above and below the
diGestive tract. It is not ~true coelom but perhaps a persistent cleavage cavity .
7. Extending from tho hinge lin0 to tho velum nrc several strands of muscle
fi bros.
3.
- 10-
IV.
Tho Trochophorc S tcgo.
Study the Trochophore st~e of Cumingia (12-17 hrs. after fertilization) and
its transformation into the Vellger larva, (lS-24 hrs, after fertilization).
The trochophore is of short duration, (less than ten hours). It is best
studied 12 to 17 hrs. after insemination. Mount some trochophores in a light suspension of india ink, and entangle them in shreds of lens paper.
1.
2.
3.
Observe the general shape in side and polar vieus.
Position of the band of cilia,
Apical tuft?
None of the internal organs can be seen because they are
Draw side and pol.1.r vie\7S in outline, showing ciliation,
Litarature!
obscur~d
by yolk,
Grave, B. H. 1927. Natural History of Cumingia tellinoides,
Biol. Bull, vol. 53.
Morgan, T, H, 1910, Cytological Studies of Centrifuged
eggs, J. Exp, Zool, vol, 9.
REGENERATION IN TUBULARIA
In Tubularia we have a system in which each part is able to reform or reconstitute into a differentiated hydranth such as develops normally from the egg.
When the stern is cut through, the opening closes and the stem cells are converted
directly into a hydranth without further cell division. There are, therefore, no
special reserve cells which supply the cells for tho hydranth nor any outgrowth
from the cut surface, Thus, a hydranth can form in two entirely different ways:
1) by cell division of tho egg with the orderly fornation of a larval stage (the
actinula) and 2) by a direct transformation of adult stem cells into a hydranth.
By studying the second process we can separate the various embryonic processes and
deal with differentiation nlone,
The process of differentiation by definition being tho process of becoming
different we must either start ,.,1 th a system in which there are differences or
create differences in a homogenous system, Tubularia stem shows some differences
but these differences arc such that the chemical environment can override them estnblishing a neu polarity. If the stem be cut into n number of small pieces each
piece will form a hydranth at the distal end but none at tho proximal end, thus
exhibiting a polarity. However, if each distal end bo placed in a glass tube this
polarity can be reversed and a hydranth will develop at the opposite end, the
proximal , Obviously then both cut ends tend to forra a hydranth, but tho tendency
is greater at the distal end and this in some way represses the form~tion of the
proximal hydranth (Doninance).
Dominance is important embryologically as it allows for the formation of a
single structure from u mass of tissue which is capable of forming more than one
structure. The region uhich has tho greater tendency to form a structure represses
adjacent tissue fron forming tho structure. This night bo explained on a nutritional basis with the region of greatest activity drawing materials away from adjacent regions. In a crowded mass of cells as obtnined in the gastrula stage there
must be a keon competition of tho various colls for nutriments and a mutual inhibition by cells caused by their excretory products, In both of those phenomena
the more rapidly metabolizing cell has the advantage and is most likely to differentiate.
The above situation can be brought about experimentally by taking a mass
of cells nnd plncing a barrier to free diffusion on one side of the mnss. This
has the dual effect of an increase in excretory products and a decrease in oxygen
-11-
tension and the result is that differentiation is inhibited on this side while the
hydranth forms from the cells of the opposite side.
In the natural environment with the perisarc around the cells it is highly
probable that this perisarc acts as a natural barrier to free diffusion and that
the hydranth forms at the cut end because these cells are released from an inhibition caused by both excretory products and low oxygen tension.
COLLECTION AND CARE OF TUBUIARIA
Theoretically Tubularia is a solitary form but practically so many individuals grow together that a dense tangled mass usually results in the older forms.
Young short stems are tho best for experiDental ~ork and can be obtained fron floats
and rocks in swift current. In gonoral it is bost to collect your own stems. Since
the stems need running water and a low temperature (18 C) they do not keup nell in
the laboratory. In nature the hydranths drop off about tho end of July and tho
stens r emain dormant until tho water cools down in the fall. At Woods Hole the stems
appoe.r in mid Juno and can be used unt~l August. However, sin: o tho wnters of Capo
Cod bay on the North shore are much colder, Tubularia may bo obtained from tho North
end of the canal throughout August.
'.
The steQs as collected vary in length, thickness and in gonoral physiological condition (sone are crushed, some starved , others very old with large gonophoree).
Therefore, we cut the stems off and sort thon out in a large finger bowl b eing caroful not to crush them . For most work tho short stems about 10 mm in length are
suitable. Those are solocted for uniform dianetor and nppoarnnco and the hydranth
is cut off a fo~ ~from its baso. It is necessary to cut off some of the stem with
the hydranth as this part of tho stem docs not regenerate consistently, especially
in older stems .
Of course, even after selection the stems ~ho~ some variability in regeneration nnd so it is bost to p9ol all the stens for one experiment nnd select at random for tho various parts of the experiment. Thus, if you are treating tho stems
in 4 different ways you should separate the stens into 5 lots at random using one
lot as a control. Tho number in each lot depends on the nature of the experiment .
~ny experiments aro of tho all or none nature and 10 stems in each lot are suffici ent. In experiments where rate of regeneration is compared under different conditions it is best to use about 25 stens in each lot. This gives a very high nccuracy in averaging rates.
The stems nrc kept in running water and cool by placing them in syracuse
watch glasses nhich a.re first submerged in large fingerbo'.'Vls through which sea
\"later is running. Some care rust be taken that the stems are not wnshed a.wey .
The instrunonts used for cutting nnd fu~ndling are a pair of sharp scissors,
a pnir of forceps and a medicine dropper. In using the forceps care must be takon
that only tho parts which nrc finally cut off and discarded are handled. After
the stems have been cut to size they are transferred with a pipette, and must not
be h~dled with forceps.
VThen tho temperature of running sea \70.ter gets above 25 C it is necessary
to keep the stems in a refrigerated bath or regenoro.tion will not be consistent.
The Warburgs nppenr to be an excellent place for Tubularia as the constant
shaking keaps them well n!!rn.ted and bathod with solution with the result that thoy
regenerate rapidly and form largo hydrnnths.
DOMINANCE IN TUBULARIA REGENERATION
Four lots of stems 'vi th 10 stems in each lot vrill be used. 1) long stems
from lO to 15 mm in length after cutting; 2) short stems about 6 mm long; 3) short
stems ~ith a lignture tied in the niddlo of tho stern; 4) very short stems about
- 12 ...
1 or 2 o=- long. Select steos about 20 to 25 mn long for fl; for ~2, 3 and 4 stems
un to 10 on are suitable. These can then be trinned to size being very careful to
reoove sone of the stem when cutting off the hydranth.
This experioent is of the all or none nature; that is, tho proximal hydranth will usually either develop in all sterns or be absent in all stems because
0 : do~inance of the distal hydranth.
For this experiment it will be necessary only to record the n~ber of
bydranths developing in each of tho four lots . It nay be snfoly assuned that tho
distal hydranth always develops so that if ono end fails to forn a hydranth it is
nloost certainly the proximal end.
After tho sterns have been cut to size they are placed in tho watch glasses
in tho circuleting sea uatcr. Care must be taken that tho incoming sea wntor docs
not wash the stems away. Have tho sea water enter at tho bottom of tho large finger bowl and do not have tho water run too fast.
At 48 hrs . tho hydranths should bo fully f ormod within tho porisa.rc and
som8 should be enorging . Record both the nunbers. Discard stems which do not show
regeneration at either end.
EXPERildENTAL CONTROL OF POLARITY
This again is an all or none typo of experiment and only 10 sterns aro
needed in o~ch lot. Draw out glnss tubes about 1 mm in inside diameter so that tho
stems will fit into those tubes loosely. Tnke 10 stems about 6 mm long and insert
tho distal end of the sten into the tubo leaving tho proximal end free. Uso 10
controls to dctornine the behavior of tho proximal and distal ends under normal
conditions.
GRADIENTS IN TUBULARIA
Grndients are qu~titative differences which cun be detected inn variety
of ways but perhaps the most signific~t difference is n difference in the ruto at
which the stem foros n hydranth at different levels of the stem. (Accompanied
with this is a difference in size of the regenerating hydranth.) We will attempt
to ncasure the ti~e fron cutting of tho stan to the fully fornod hydrnnth. Since
the first hydranths develop nt about 24 hours and others continuo to develop at
later times it is best to start the experiment ns onrly in tho morning as possible
so as to huve all tho next dny in which to make observations ~ The stems should be
cxaninod every 2 hrs. nfter tho first hydrnnths have formed for further regeneration. The criterion used for a fully forned hydranth is tho presence of tho tontncles and especially a sharp constriction nt tho base of the hydranth whilo still
within the perisarc. Simply look at the ends of the stems under low power nnd
record the time when this construction is first noted.
Select stems about 25 mrn in length and cut off the hydrnnth together with
a few mm. of tho stem as before. Then cut tho reminder into pieces about 5 to 6
~~ . long discarding what is left.
As these pieces nro cut placo thorn in sepnrnto
dishes labelled disto.l, middle and proximal thirds. Continue 2 hr. observations
ns long as possible and tnko n final observation the second day at 48 hours and
nverngo tho timos, by totalling the total number of hours for each lot and dividirl{; by tho total number of stems. If some stcns hnvo not yet regenerated include
those in the total number as they would regenerate if given time.
REFERENCES TO Tu:BULlillL (ALSO CORYUORPHA)
Soc Biblloc raphy for Coclonterntos
for detnilod rof cr onco
Child
1941 Patterns and probloms of development. pp. 313, 333, 345, 359, 372, 378
and 413; Also Cho.pt. 8 \7hich is theoretical .
'
17 ·-
Barth
1938
1938
1940
194o
Physiol, Zool., 11:
B~ol. :Bull.,
74:--
B1ol, Bull., 1[:
Biol. Reviews., !2:
Rose and Rose
1941 Physiol. Zool., 14:
Joore 1939
Biol. Bull,,~:-­
Miller 1937 Biol. Bull,, TI.:
~oog and Spiegeloan
1942 Proc. Soc. Exp. Biol. Med., ~:
!.1oog
1941 Biol. Bull., 81:
Goldin 1942 Biol. Bull,,~:
Zwilling
-1939 Biol. Bull., Ii:
1944
COELENTERATES
Introduction to Hrdrozoa
a)
Sensitiveness of the Material.
While working with Coelenterates in the laboratory it is essential to remember that the hydroids are very sensitive to environnental conditions. They do not
survive well in the laboratory even in aquaria of running water. Do not crowd either
the adult steos ~r tho· oggz ~d enbryos. In general, your procedure uril
to look
over a good-sized colony of hydroids under the lowest power of nagnification, and
to clip off a feu pieces containing the best embryological material. These can bo
rinsed in running sea water, and segregated in plenty of sea wator in a dish for more
detailed study.
oo
b)
Types of Life History.Illustratod.
There are two phases of embryology in the Hydrozia. Characteristically,
a long series of asexual reproductions (by budding nnd other methods) is interrupted
at irregular intervals by isolated examples of sexual re~roduction. This alternation
of sexual and asexual generations is closely paralleled y an nlternation of structure, since zygote production is usually ncco~lished by medusae nnd the production
of buds by polyps .
In some hydrozoa no medusa-form is known, in others no polyp form is known.
and there nrc nll stages between. The hydrozon available at Woods Holo illustrate
well the structural vnriability of the medusa-state. The best known exanple of a
complete medusa with a degenerate polyp stage is the idealized jelly-fish Gonionemus, but it is unfortunRtely now nearly extinct at Woods Hole. The gonorn picked
for study nre arranged in tho order of diminishing completeness of tho medusa form,
the first having free-swimoing medusae and the last mero sporosacs. The utter dogeneration of the medusa-form is illustrated by the familiar Hydra.
c)
Order of Study
Bocause of oensonnl variations and unpredictable fluctuntions in the Coelenterate fauna. some of the material described may not be available, nnd tho order
of study will have to bo announced.
Labotntory Procedure
A.
Study of Forms with Perfect Medusa
Examples:
'
I
Bougainvillia, Obelia, Podocoryno .
-
14 ·-
Characteristic lifo h~story: Zygote shed from medusa; development to planula
larva; metamorphosis to polyp; asexual multiplication of polyps by budding,
which produces a colony; nedusae formed by special buds, in a gonosome (Obelia)
or separately (Bougainvillia); shedding of nedusae, which mature slowly as
separate individuals oefore forming eggs or sperm.
BOUGAI iVILLIA.
(June, July, August; not always available in Juno)
The gonophores aro borne singly or in clusters on tho main stem and branches,
and in this genus develop into complete medusae. Tho medusa-buds arc scattered irrogulnrly throughout the colony, thoro being no orderly arrangement according to ngo.
Select buds that show various stages of medusa development and mount thorn und r covor
slips, and study their unfolding structure. Draw off water from under the covor
slip with absorbent pcper to produce a slight pressure on tho buds. Sketch throe
stages in medusa development.
When all of its parts except tho gonads nrc fully formed, the medusa brooks
loose and swims uwo:y. It lives independently one or t\70 months, the gonads gro.dunlly
mnturing. Find a well-developed specimen that is swimming actively nnd sketch its
diagramnatic structure. Identify manubrium, rndinl nod circular cnnnls, velum, ornl
tentacles, 4 groups of nargi~l tcntnclos.
Illustrations of medusa in Horgitt,
development in Goette 1 07. Also Hyonn •4o.
OBELIJ~
c. w.
101,
1 04;
Nutting 1 01, of medusa
(Juno, July ,August)
Tho gonosoocs arc several tines ns large ns the hydranths. ~~inc specimens and see if they are located at rnndom nlong the stnlks, or in rcgulnr places.
An Obelin gonoso~e has an enlarged transparent covering, the gonothecn,
with a bl~stostyle extending thru it from bnse to tip. Tho outer end, or tip, of
the blnstostyle cxpnnds to make a loose plug for tho gonothcca when mature.
1~ cluster of gonophoros is borne on the blastostylo inside the gonotheca.
Tho gonophoros nature as medusae, and brenk loose, escaping to tho outside past the
blastostylc plug. They arc commonly caught in tow nets. Their free-swimming life
lnsts two months or so, tho gonads maturinG slowly.
The older gonophores should show developing tentacles, when pressed
slightly under a cover slip. Which gonophores on a blastostyle are oldest?
With needles, pres~ on the gonothecn of a well-natured gonosome and examine under high magnification the gonophores that arc released. Those that nrc
oldest ony sho~ swimming movements. Younger stages of development may be tensed
out from the gonosome and studied. Sketch sovernl stagos.
If ripo Obelia coloni es are kept for an hour or two in a dish of sea water
on the desk (remember not to crowd them), swimming medusae can usually be detected
inn good light 1ith the n0ked eye. Exa~inc some of them when available, and note
thnt their structure is slightly imperfect cor~ared with that of Bougninvillia.
The velum is reduced to a narrow and somewhat lobed membrane near tho
bnsos of the tentacles. This nn.kes possible nn eversion of the bell uhen tho
medusa comes to rest, so that the manubrium sticks out froQ the center of tho convex side, like the handle of n post-hurricnne unbrelln. Wntch tho swimming movements, and see how this hnppons. In the everted condition, the manubrium is still
morphologically sub-umbrollar, though this term has lost its appropriateness.
The newly shed medusa of Obolin goniculata has 24 tentacles, while that of
Obelia commissuralis has 16. Both forms may be available in the laboratory. Neither
has gonads developed at this stage.
-
11) -
Illustrations o!' Obelia e:.1bryolo.EV in
~rnan
1
4o, Goette 107.
(June, July. 1 or 2 colonie3 will be collected with each 100 Hydractinia colonies fron Sheep-Pen Harbor. None from Pasqua.)
The highly specialized colony grows in an encrusting nat on snail shells,
etc., and is aloost exactly like Hydr~ctinia. Both ~~ve three types of individuals:
feeders, gonosoces, sti~ers (Cf. descriptions of Hydractinia below, p. 7). Podocor}~e is included here ecause of its startling o tngenetic contrast to Hydrnctiniu.
Medusae of Podocoryne are nearly perfect, and oo.y produce scvcrfl.l gon rations of now medusae by asexual budding bofore getting around to th ir onin businoss
of gaoete production. Cf. Goette, '16. (The sporosacs of Hydractinin. bonr very
little rcsonblanco to oedusae, being highly degenerate.)
Sketch the three types of polyps ond show several stngos in nodus~ dovolopcan be onde out along the rndial cnnals of tho swimming ncdusne, vor.y
iw~~ture but scxunlly distinguishable.
The nsoxunl colony gives off oith r rnnl or
female neduscc, not both.
ment.
B,
Gon~.ds
Study of Foros with Imperfect Medusae.
Examples:. Pennaria, Tubularia.
Life Histories: Zygote shed from short-lived imperfect medusa. (Pennaria)
or retained in reduced sessile medusa form (Tubularia); development to
planula larva and metamorphosis to ~olyp (Pennaria) or development to
Actinula larva and growth to polyp (Tubularia); a sexual multiplication
of polyps by budding to produce colony; gonophores formed by specinl buds
on hydrnnths; maturation of gonophores (imperfect medusae) and fertilization either in situ (Tubularia) or within the limits of the colony
·
during their detachment (Pennaria).
P~~TARIA
(July, August, September; begins to ripen middle of July.)
Gonophoros bud off singly around the lower portion of the hydranth. They
form slightly reduced medusae with rudimentary tuft-like tentnclos. Before opening
out ns trnnspnrent bell- shaped forms they suggest cocoanuts. A single colony boars
gonophores of one sex only , but in the living individuals, sex can be diagnosed only
with difficulty until they mature , when the pinkness of eggs and the whiteness of
s:J;Brm appear . (Smallwood 1 89) . 11 Ma.le" and 11 female 11 colonies are actually asexual.
bearing male and female gonophores respectively.
Maturo Pennaria colonies festooned with shedding medusae provide nn astonishing and beautiful spectacle that every student should certainly see. Tho material
ripens in the season of warmest water, and a demonstration of the shedding will be
made when possible . It starts early in the evening and continues through midnight.
It is usually best seen in material brought into the laboratory the preceding day
(i.e . , the second night) .
The ripe medusae gradually start a rhythmic twitching. Those which are
males emit puffs of whitish sperm , and those which are t amales eject with greater
travail the three ·to six opaque pink eggs . In the south, Pennaria medusae generally
break loose from the colony and swim about during this discharge, but at Woods Hole
they generally remain attached, and the eggs may not be ejected until long after
f ertilization. The medusae finally drop off, swim very feebly if at all, shrivel
rapidly and die in a few hours.
'
Put small selected stems from ripe "male" and 11 female 11 colonies together in
a fingerbowl after careful rinsing, at 3-4 p . m. and leave them overnight. They are
extremely sensitive to overcrowding ! Next day, remove the stems and look with naked
- 16 -
eye for free medusae
eggs.
~s
evidence of ru odding.
If they aro found, look for developing
~e eggs are very simple and slightly ameboid, ~ith no apparent membranes.
Follow their development as far as possible. If ~~e water is changed several times,
the planula stage should be reached in 24 hours, and stages in the very simple metamorphosis to the polyp form may be observed. (Hargitt, G. T. 1900, 1909).
Tease apart male nnd female gonophores and examine the eggs and sperm.
Sketch several stages in development of the medusa; sketch the mature male and
female medusae; sketch several cl~vage stages if found.
Illustrations of medusa development in Goette 107; of cloavag
C. VT., '00. Also Hym.:m 1 4o.
TUBULARIA
in
H~rgitt,
(June, July.)
The gonosomes of a well-matured specimen form long racemes or clusters of
gonophores crowding and drooping from the region between the two circles of tentacles
on a hydranth. The gonophores are quite severely reduced medusae which never become
free-swimming, usually have no evident radial or circular canals and develop nothing
but buds for tentacles. Uale and female gonophores occur in separate colonies.
Diagnosis of sex by sight is impossible in the immature, but gonophores that contain
embryos are easy to tell from thoso that are filled with a cloudy mass of sperm.
Early stages of developing embryos arc found by tonsing with neodlos, those noar tho
hatching stage are visible in situ.
Examine a ripe male gonophore, considering it as a very degenerate medusa.
Notice and sketch its mode of attachment, its shape, the structure of its free end
and the position of the sperm surrounding the dark red manubrium. Crush it on a
slide and inspect the motile sperm under high power.
Examine and sketch a ripe female gonophore.
distal end appear only as four short blunt knobs, but
times slightly elongated. The eggs come from favored
swallow up their neighbors, lying in the space around
Usually the tentacles at its
one or more of them aro someoocytos that progressively
the spadix (manubrium).
When ripe the egg is very large and somewhat irregular in shape (Allen '00 ,
Lowe '26). After fertilization, cleavage is often chaotic, apparently either a
coeloblastula or n morula may be formed (Lowe 1 26), and gastrulRtion of the former
has been described (Benoit 1 25) as n mixture of dolaminntion and multipolar prolifere.tion. Tho embryos are developed up to the 11Actinulo. 11 stnge within the gonophore.
The Actinula larva is to be considered as a precociously metamorphosing form, part
planula and part polyp.
By teasing some female gonophores open with needles, collect and sketch eggs
and larvae in various stages. The larvae just taking shape are flattened with blunt
marginal processes. At first these are not symmetrical in outline but radial symmetry is attained later. When tho larva has reached the Actinula stage it has a
mouth and aboral tentacles, and a rounded aboral body that later becomes attached
at its tip.
Study and sketch tho structure of a fully formed Actinuln larva. Look over
some ripe 11 female 11 colonies for Actinulo.e beginning to escape from the gonophoros.
Actinulao will not develop further without feeding.
Tubularia anatomy and development is illustro.ted in tho texts of lAacBrido
Korschelt '36 and Hyman 1 4o. For cleavage cf. Allen 100 , Hargitt, G. T., '09.
For gonophore development, cf. Goette 1 07.
1 14,
- 17 -
c.
Study of Foms with Degenerate Hedusaa.
les
onotheca:
~~p~l-e-s-w~i~tr-~~~~~~~~~~ny~d~r~a~c~t~1~nia,
Cam anularia, Gonothyrea.
Eudendrium.
Lffe Histories:
(a) C~nnularia, Gonothyrea; Zygote develops into planula larva
inside sessfie deger.erate medusa; planula esca. es, lives free awhile,
~etamorphoses into a polyp; asexual multiplication by buds; colony
formation; degenerate medusae (gonophores) formed on a blastostyle;
gonophores mature in situ; sperm are shed, oggs fertilized in situ.
(b) ~dractinia, Eudendrium; Zygote develops into planula larva
either ins o gonosom'o (Eudendrium) or after being shed from gonosome
(Hydractinia); planula metamorphoses into polyp; asexual multiplication
by buds; colony formation; gonosomos formed from hydrorhizn (Rydnctinia)
or by transformation of hydronths (Eudendrium); gonophores (highly
reduced medusae or sporosncs) borne on gonosomes; oggs and sperm
formed in the sporosacs; ~s fertilized in situ (Eudcndrium) or
during shedding (Hydrnctinia.).
C.AiAPANULARIA
(June, July).
It is not safe to try to distinguish this genus from Obelia by the anatomy
of the feeding individuals. Even the gonosomes are similar in appearance in the two
genera, each consisting of a transparent gonotheca with the blastostyle extending
from base to tip and gonophores budding from it.
The striking difference is that Obelia produces nearly perfect free-swimming
medusae, whereas Campanularia produces gonophores so utterly degenerate that their
medusa-like structure can on1y be mado out in sections. Each gonophore on the blastostyle of a "female" colony contn.ins a very largo irregularly shaped ogg which is
fertilized in situ, cleaves, forms a morula, gnstrulatos by delnmination and reaches
the free-swimming planula stage, still in situ. Campanularia therefore releases
from its gonotheca not medusae but planulae.
Because the gonophores are so inconspl.cuous and the embr,10s so obvious, the
colonies which produce female gonophores and later contain embrybs are loosely
spoken of as 11 female 11 colonies, although they are asexual.
Select from a "female 11 colony a gonosome showing eggs in the basal gonophores. Mount it on a slide, study with various magnifications under the microscope, and sketch it.
Select and sketch another gonosome with planulae showing near the tip.
Squeeze or open the gonotheca with needles and liberate the planulae. Notice their
ciliated ectoderm and watch their movements. If tho planulae are well matured, they
are two or three times longer than broad. They show maggot-like movements oven
while within tho gonotheca.
Carnpanularia is very favorable for the study of planula metamorphosis.
3 to a watch glass,)
and cover them. They should attach to the glass in 4 to 10 hours. Eo.ch should then
open a mouth, bud out tentacles, secrete hydrotheca and perisaro, and become a fully
foroed individual polyp in two or three days. When the plunulne have ntt~ched, the
water should be ch~ed in the dish at least twice a day. Sketch several stages of
attc.chment and metcmorphosis.
~t a few mature planulae aside in sea water (not more than 2 or
The gonosonr.es of 11 male 11 colonies are similar in form to tho female. The
gonophores, when mature, arc rounded and have a thin milky-gro.y color. The sperm
become nctivc when they nre discharged into contact with soa wnter, as mny be seen
by crushing a male gonophore under a cover slide while watching it under tho microscope.
·.
For illustrations of Campanularia gonophore development cf. Goette 107.
GONOTHYREA.
(July , August) •
As in Obelia, the medusae develop within tl:;le go no theca. Uhen rna ture, instead of swimming away they r emain attached to the end of the blastostyle, projecting outside the gonotheca in groups of three or four, like toy balloons, Within
the balls of the medusae, the eggs of 11 foma.le 11 colonies are fertilized by tho shod
sp~rm from 11 role 11 colonies,
The zygotes develop to tho planula st~e before being
set free, after which the medusae drop off.
Sketch planulae in various stages of development in the projecting attached
!::ledusae of e. ripe 11 female 11 colony. (Wulfert '02).
Goette
Gonothyreo. cleavage illustrated in Wulfort '02~ medusa. devolopr.10nt in
H~ •4o and Kirscholt •36.
'07. Cf, ulso texts of
EYDRAC~INIA.
(Juno, July, August)
Colonies of this form are fairly common on Littorina snnil shells inhabit ed by tho s~all hermit crab, Pngurus. There are three typos of individuals in the
fully developed colony: ordina.ry polyps (feeders), throadliko coiling forms with no
nouth nnd un apical knob of nemntocysts (stingers, cornnonost a.round tho lip of the
shell), and gonosomes. The three types alrarise from n hydrorhiza network covering
a rust-red sp ino-studded crust,
a)
Gonouomes and Gonouhorcs:
The gonosomes or reproductive individuals arc usually without tentacles
and have a large knob of nonatocysts on tho proboscis; each bears a nunbor of gonophores, ;1hich are medusa.-buds reduced to tho status . of sporosacs. Ripe 11 r.mle 11 o.nd
11 femnld 1 colonies can be told apart with the naked eye since the eggs within the
sporosacs are dull green against the red hydrorhiza, and the sperm when ~ature are
a white ress.
Remove several 11 nale 11 reproductive individuals showing ripe sporosacs, and
crush tham slightly under a coverslip on a glass slide. This may be dono by drawing off some of the water vii th blotting paper. The sporosacs fall far short of
being perfect medusae. Note stages in the production of sporn within thorn. Burst
a nnturo suorosac and study the sper~ under high power.
Sinilarly , crush several "female" reproductive individuals, and observe
the cge s with thei r largo germinal vesicles, in various stages of development.
Sketch tho 11 femalo 11 gonosoLie with its feoolo gonophores nnd contained eggs.
b)
Clea.v~ e
nnd Development.
If a number of 11 malo 11 and 11 fomalo 11 colonies of Hydrnctinin. nrc put together in ~ large dish of sou ~ator (or a pair of prime colonies in a fingerbowl) end
l ef t overnight, eggs should be shod and fertilized between 7 and 9 a , m. WST. The
shedding ca.n bo controlled by light. If fertilization rnd cloavngo stages ~ro
needed l~tcr in tho da.y or in the evening, tho colonies no.y be kept illuminnt cd
during tho preceding night, put i~ the dark for n couple of hours and roilluminated
one hour before the tine \7hcn shedding is desired. Eggs nrc shod in 55 minuteG,
spe r~ in 50 minut es . Cf. Ballard, 1 42.
lAntcrinls for a study of the entire development of Hydrnctinin from egg
to polyp Tiill be made available. If possible , observe the shedding of e~gs and
sperm. Sketch eggs undorgoing first throe clonvnges, elonGated Gnstruln , swinming
and a.ttached planulae, metamorphosing form and yoUQg polyps . ·
Eggs o.re heavily yolky and usually green , but occasionally gray , ornngc
or pink. ~~turation takes place during the half h ur preceding he ding and polar
bodies are lost soon after, a loose jel~r being the only covering of the egg.
Cl~vage mny be irregular or even chaotic, but usually the slightly ameboid egg undergoes thre8 equal tot~l clon•~bes, ~~ch at right angles to the proceeding. The separating .~irs of blnstomeres tend to retain broad protoplasmic connections \'lith onch other on the side op:t;osi to to the clenvogo furrow, until just before
the succeeding cleavages begin. It is soon apparent that thore is much variation
in the ti~o ~nd dogroo of shifting of positions of tho blasto~ores, ut the extraordinary and Chaotic cleavage patterns co~only seen in tho classroom nro often tho
result of drying up n.nd concentrn.tion of sea water, or other unfavorable circwnstnnc s.
!Aitotic synchronism quickly disn.ppcars. Gastrulation is snid to st rt oven
as early as tho 16-coll stnge, by ~ixod del~inntion and nultipolar proliferation.
Tho gastrula loses its spherical for~ and spends a few hours ns nn irregularly bunpy
oblong nnss, then returns to tho spherical foro and gradunlly lengthens into the
planulc. foro.
At the end of 24 hours the er:lbryo is a 11 prep1Mula. 11 (Toissier ' 27) with an
elonGated ovoid form, recognizable polarity and cilintion which enables it to swim
heavily. During several days it lengthens, one end becoming slimncr nnd liomor,
while it rolls and crawls nlong the botton like n plannrinn. Tho biG end which
goes first in this novemont is the end which later produces tho ndhesiv~ disc by
which it attaches for metamorphosis. It beconcs tho aboral end of tho polyp.
Following attachncnt of the attenuated planula, there is a dolny of a fow
hours to several dnys, c.nd then tho tapering free end shrinks down nlnost to tho
substrate, where it shortly produces a mouth and a succession of tentacles. The now
polyp elongates, its attached end meanwhile actively Eonding out a number of nnastomosing and encrusting hydrorhiza processes fro~ which sprout now polyps.
Illustrations of clqavage in Bunting '94, Beckwith 1 14; of later planula
development in Toissior 1 27; of developing gonosonc in Goette 1 07 and 1 16.
EUDENDRim~
(July, August).
~no gonosones of this genus are degenerate sessile medusa-forms or gonophorcs, strikingly different in the two sexes, borne at tho bases of special hydranths
which lose· their tentacles and degenerate while the gonophores are ripening.
11 Ferole 11 colonies bear loose irrOt,~lar tufts of sporosacs attached to tho
sterns, each ripe sporosac beint; bright orango in color. 11 M11lc 11 colonies bear li ght
pirik sporosacs arranged in groups of two to four or moro in a lino, the lines radiatinG fron a co~on point on the b11sc of the degenerated hydranth. The ripest nale
sporosacs occur at the periphery of tho cluster and are white with sperr:l, Sketch
both malo and fe~lo sporosacs.
Eggs are fertilized within the fenu-,le Gonophore or sporosac , nnd develop
to the planula stage before boing lib ern ted.. (Hargi tt, c. VT., 1 04) . Teas u out embryos from different colonies and study all star,es found. Tho C{;B S r.r e so ri ';n
in yolk tho.t they cleave like insect eggs, The gastrulation is by an ext.r-cl.r,rrlinn.ry
syncytial delamination.. Metamorphosis is simple .
Eudendrium cleavag e illustrated in Hargitt 1 04 (Zool. Jf'.h:-b.) .; Conon:)me
developr.10nt in Goette 1 07 . Cf. ~lso texts by Korschcl t '36 and Hyr..::m '·10 .
Development of Scyphozoa.
AURELIA OR CYANEA.
(April - Jw1e)
Both thes e jellyfishes ho.vo oral lobes extending downwarcl. :Cn na tnr-J
specimens granular I:lnterial will be found entanGled on the lob es or conta in ed in
- 20-
5 ~11 brood sacs in the lobes. Tease off sooe of this caterial into a drop of sea
water on a slide, and examine under the ~icroscope. Eobr.yos of different stages can
be found, froc spherical cleaving eggs to oval r,astrulating forcs and fully forned
stocky, active planulae. (Hargi tt, G. T., 1 09) (Rein, 1 00). Is cleavage regular?
Sketch the embryonic stages that are avoilablo.
Select a nunber of active planulae nnd place then in clean wntch glasses
of sea water for further study on later days, Their gradual change in form, attnchoent to the botton, acquisition of tentacles and elongation into the sessile scyphula
The rcsenblance of tho scyphula to n simple polyp is
6 tage can be easily followed.
obvious. Attachoont plus the formation of 2 to 4 tentacles occurs on tho second dny,
as does the development of an open mouth. There are 8 tentacle' nt 4 days, 16 at
2 weeks, 24 at 1 oonth. Sketch the sc3~hula in side vio~ nnd in top viau.
The scyphula or scyphistornn stage lasts throughout the winter. Tho aninals increase in size and undergo asexual reproduction by transverse fission into
ephyrae ( 11 strobilizntion 11 ) , And by other methods (Percival '23). The tiny ephyrne
(larval jellyfishes) are liberated and gradually transform into the adult form ovor
a period of many months. See demonstration slides of strobilization and ophyrno.
Aurelia life history illustrated in the texts of Uac::Brido
136 and Hyna.n 1 4o •
....
1.. 4 ...
,·•
- 21-
1
14, Korscholt
1944
BIBLIOGRAPHY
G~...RAL
VTORKS AND COMPARATIVE EilBRYOLOGY OF
PROTOCHORDATES
I:'
Conklin, E. G.
1932. The embryology of Amphioxus. Jour. ldorph., ~:69-149.
1933. Development of isolated and partially separated blastomeres of
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Dalcq, A.
,
1935. L 1 0rganization de 1 10euf chez les Chordes. Paris.
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1928. Traite dfEmbryologie comparee des Invertebres. Paris.
Herdman, E c.
1924. Botryllus. Liverpool Marine Biological Committee Memoirs, XXVI:
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1899~
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Korschelt, E.
1936. Vergleichende Entwicklungsgeschichte der Tiere. Jona.
Seeliger, o.
1893
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Willey, A.
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TUNICATE EMBRYOLOGY
Borrill, N. J.
1929. Studies in Tunicate Development, I. General Physiology of Development of Simple Ascidians. Phil. Tr. Roy. Soc. London, 218 (B):37-78.
1931. I I. Abbreviation of Dovolopment in tho l.iolgulidc.'l.o, Ioia:-;(B) :281-346 •
• Differential Retardation and Acceleration. Ioid., 225
1935.
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1932. Mosaic Development of Ascidinn Eggs. Biol. Bull •• §1:381-386,
Chnbry, L.
1887. Contribution a 1 1 ombryologie nor~le et teratologique des Ascidies
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Cohen, A. & Bcrrill, N. J.
1936. The early development of Ascidian eggs . Biol. Bull,, ~:78-88.
Cohen, A. & Berrill, N. J,
1936. The development of isolated blustomeres of the ascidian egg. Jour.
Exp. Zool., 74:91-118.
Conklin, E, G.
-1905a. Organization and cell-lineage of the Ascidian egg. Jour . Acad. Nat .
Sci, (Phil~delphia),!} (2nd series) : 1-121.
1905b. Mosaic development in ascidian eggs. Jour. Exp. Zool. , 2:146-223.
1905c. Organ-forming substances in the eggs of AscidiAns, Biol . -null .,
8:205-2)0.
1911. The organization of the egg and the development of single blastomeres in Phallus in. Jour. Exp. Zoor. , 10: 39Y..4o7.
1931. The development of centrifuged eggs of AScidinns. Ibid., 6o:l-120.
Kingsley , J. s.
1883. Some points in the dovolopmont of Molgula. Proc , Boston Soc. Nut.
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1871. Weitere Studien uber die Entwicklung dor oinfachen Ascidian . Arch.
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iif
-1-
-2-
i.iorgo.n, T. H,
1938. The genetic nnd physiological problocs of self-sterility in Ciona.
Jour. Exp. Zoolp, 78:271-334.
1941. Further experiments-in cross- and self-fertilization of Ciona at
Woods Hole and Corona del Mar, Biol, Bull,, 80:338-353.
1942. Cross- and self-fertilization in the ascidian~tyela, Ibid.,
82:16].-171.
1942. cross- and self-fertilization in the ascidian Mo1gula Mahattensis.
Ibid,, 82:172-178.
1942. Do spermatozoa penetrate the membrane of self-inseminated eggs of
Ciona and Stycla? Ibid., 82:455-460.
1942. The genetic and physiological problems of self-sterility in Ciona.
V, The genetic problem. Jour, Exp. Zool,, 90:199-228,
1944. Tho genetic nnd physiological probloms of self-sterility in Ciona.
VI. Theoretical discussion of genetic data. Ibid., ~:37-Go.
Rose, s. U.
1939. Embryonic Induction in the Ascidia. Biol, Bull., II:216-232,
de Selys-Longchnmps et Damas, D.
1901. Recherches sur le development post-embryonnairo ot l'Anatomie
definitive do Molgula ampulloidos. Arch de Biol., ~:451-469.
Tung, Ti-Chow
1932. Experiences de coloration vitnle sur 1 1 oouf d'Ascidiella asperse.
Ibid., 43:451-469.
1934. Rechercnes sur los potentialites des blastomores chez Ascidielln
scabra. Experiences do translocation, do combinaison et d 1 isolemont
de blastomores. Arch. d 1Annt, micro., ]2:381-410,
Tung, T., Ku, s. and Tuny, Y,
1941, The development of the Ascidian egg centrifuged before fertilization.
Biol. Bull,, 80:153-168,
Tffi~ICATE
ASEXUAL REPRODUCTION
Berrill, N. J,
1935. Studies in Tunicate Development. IV. Asexual Reproduction, Phil,
Trans, Roy. Soc •• London, 225 (B) :327-379.
194la. The development of the bud in Botryl1us, Biol. Bull,, 80:169-379.
194lb. Size and morphogenesis in the bud of Botryllus. Ibid.,~O:l85-193.
Brien, P.
1927. Blastogenese des Tuniciers. Arch. de Biol., }l:l-46.
Cau1lery, M.
1909. Contributions a 1 1 Etude des Ascidies composces, Bull, Sci, Fr. Belg.,
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Kowalevsky, ~·
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Pizon, A,
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Ritter, ~. E,
-.
1897. Budding in compound ascidians, based on studies on Goodsiria and
Perophora. Jour. Morph. • 12:149-238.
do Selys-Longchamps
-1916, Sur le bourgeonnement des Polystyolines, Bull. Sci. Fr. Bolg.,
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Spek, J.
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Clavellina lcpadiformis, und die Bedeutung besondorcr 11 onmipotentor 11
Zcllelemento fur diesc Vergange. Roux' Archiv., 111:119-172.
•·
-3TUNICATE RIDENERATION, ETC.
Bcrrill, N. J, and Cohen, A.
1936. Regeneration in Clavelina lepadiformis. Jour, Exp. Biol. !}:352-362.
I' Brien, P.
I,
1933. Regeneration thoracique chez Archiascidia. Bull, Biol. Fr, Belg,,
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1
~:100-124.
Driesch, H.
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-Huxley, J, s.
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Perophera. Quart, Jour, lHcr, Sci., 65:643-697.
1926. VI, Reduction phenomena in Clavellina-lopadiformis, Publ. Stnz,
Zool, Napoli, I:l-35.
TUNICATE TADPOLES AND THEIR METAMORPHOSIS
Grt:>.v e , C.
1920.
Amaroucium constellatum. I, The activities and rea c t i ons of the tadpolo larvue, Jour. Exp. Zool,, 30:239-258.
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Morph, , 3.§.: 71-101.
1926. Molgula citrina, Activities and structure of the free-swimming larva,
Jour, Morph. and Physiol,, 42:453-471.
~935- I~etumorphosis of Ascidinn lnrvae.
Papers from the Tortugns Laboratory, ~:209-292.
Grave, c. and Nicoll, P. A,
1939. Studios of larval life nnd metnmo~hosis in Ascidia nigra nnd species
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Grave, C. and Riley, G, .
1935. Development of the sense organs of the larva of Botryllus schlossori,
Jour, Uorph,, 57:185-211.
Grave, C, and Woodbridge, H.
.
1924, Botryllus schlosseri: the behavior and morphology of the free-swimming larva, Jour. Morph. and Physiol,, Jt: 207-247.
Kowalovsky, A,
.
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Petersburg,, ~:
Mast, s. 0,
1921, Reactions to light in the larvae of tho ascidians Amaroucium constollatum and Amaroucium pellicidum with reference to photic ori entation,
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van Benedcn, E, and Julin, c..
.
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BIBLIOGRAPHY FOR COELENTERATES
1944
MONOGRAPHS, KEYS, SPECIAL VTORKS
Brooks, W. K.
1886, The life history of tho Hydromodusao: A discussi on of the orie in of
tho medusae, and of tho significance of mctngenosis, Memo irs, Bos t on
Soc. Nat, Hist., 1:359-430.
I"'"
-4F8yn, B.
1928.
Goette, A.
1907.
Ha.rgit t, c.
1901,
Studien dber Geschlecht und Geschlechtszellen bei ~·droiden,
Bemerkungen tiber die Entstehung der K imzellen und die Entwicklung
der Gonophoren bei Clava squamata, Roux 1 Archiv,, 114:501-510.
Vergleichende Ent~icklupg~geschichte der Geschlechtsindividuen der
HYdropolypen, Zeit. f, wiss. Zool,, ~:1-336,
VT.
Synopsis of North American Invertebrates, XIV. Tho Eydromedusae.
Ao. Nat., }2:301-315, 379-395, 575-595.
XIV. The Scyphomedusae. Ibid,, 11=331-345.
The medusae of the Woods Hole region, Bull, Bur. Fish., 24:21-79.
1903.
1904.
-Ha.rg itt , G, T,
Germ
cells
of
Coelenterates,
VI.
Gonoral
considerations,
discussion
1919.
and conclusions, Jour, Morph., TI.:l-60,
Hyman, L. H.
194o. The Invertebrates, I, Protozoa through Ctonophora. McGraw-Hill.
Kingsley, J. S,
1910. A synopsis of the fixed hydroids of Now England, Tufts College
Studies, l:l3-38.
Mayer, A, G,
1910. !Aedusae of the world, I. and II. The Hydromedusao, III. The
Scyphomedusae. Carnegie Inst. Washington, 109:
Nutting, c. C,
1901, The Hydroids of the VToods Hole region, Bull, u. s. Fish Commission,
!2: 325-386.
Sumner, F. G. et al
1913. A biological survey of the \Taters of Woods Hole and vicinity. Bull,
Bur. Fish, , .n_:
PAPERS ON HYDROZOA
}..
GENERAL ELffiRYOLOOY, D::£SCRIPTIVE
Allen, Carrie 1..1.
1900, A contribution to the development of Parypha (Tubularia) crocea. Biol,
Bull,, 1:291-315.
Beckwith, Cora J, 1914. The genesis of the plasma structure in the egg of Hydractinia echinata. Jour. Morph,, ~: 189-251.
Benoit, P,
1925. L 1 ovogenese et les premiers stades du dcveloppement chez la Myriothele
et chez la Tubulaire, Arch, de Zool, Exper,, 64:85-326.
Bunting , Uar tha
~894.
The origin of sex cells in Hydractinia and Podocoryno, and the development of Hydractinia, Jour, Morph., 2_:203-236,
Doflein, F,
1896. Die Eibildung boi Tubulnria, Zeit. f. wiss. Zool,, 62:61-73,
Goette, A.
1916. Die Gattungen Podogoryne, Stylactis und Hydractinia, Zool. Jahrb,,
Abt. System, Geog. Okol, Tierc., ~:443-510.
Gr8nberg, G.
1898, Bcitrtlge qur Kenntniss dcr Gattung Tubulnria. Zool. Johrb. Abth, f,
Annt,, ll:61-76.
H~rgitt,
c. w.
1900.
1904.
-
A contribution to the natural history and development of Pcnnnrin
ti~rolla, Am. Nat., ~:387-415.
The early development of Eudondrium. Zool, Jahrb., Abt. Anat. u. Ont,
20:257-277.
Hnrgitt, G. T:1909. Maturation, fertilization and segmentation of Pennaria tiarella and
of Tubul~rin crocea. Bull, Mus. Comp, Zool, Harvarc, 21_:159-212.
-5Lowe, E.
1926.
The m::lbcyo1ogy of 'l'ubularia
1.2_:599-625,.
larynx~
((u..'"'.rt. J ur. ·l!.icr. Set. ·,
!lay, A. J,
1903.
A contribution to the morphology and development of Corymorpha pendula Ag, Am. Nat,, l[:579-599.
Smallwood, VT. lA.
1899. A contribution to the morphology of Pennaria tiarella. Am. Nat.,
33:861-870.
Smallwood, VT:I.i,
1909. A re-examination of the cytology of Hydractiniu and Pennaria. Biol.
Bull., U: 209-24o.
Teissier, G,
1926. Notes critiques sur la morphologie des gonophorcs choz los hydrniros.
Arch. de Zoo1, Expor. ot Gon,, §2:75-86.
1931. Etude Experimentale du dev oloppcment do quelques hydraires Ann. dos
Sci. Nat, Zool., 14:5-6o.
Tcissier •. L. nnd G,
1927~
Les Principales etapes du daveloppement d 1 Hydractinia echinAtn. Bull.
Soc. Zool. Fr., ~:537-547.
VTu1fort, J.
1902. Die Embryonale Entwicklung von Gonothyraon loveni. Zeit, f. wiss.
I
Zool,, ~:296-327.
B.
SPECIAL TOPICS - HERIJJ.J>HRODITE GONOPHORES, NORMAL AND EXPERIMENTAL
Bunting, Martha (listed above)
F8yn, B,
1924. KHnstlich hergestellter Hermaphroditsmus boi Hydroiden. Zool, Anz,,
61:105-110.
1927. !fudien uber Geschlecht und Geschlechtszellen bei Hydroiden. I, Ist
Clava squamata (Uu11er) eine gonochoristische oder hermaphrodite Art?
Rowe' Archiv., 109:513-534.
·
1927. II. Auspressungsversuche an Clava squamata (Muller) mit mischung von
Ze11on aus Polypen desse1ben oder verschiedenen Gesch1echts, Ibid.,
110: 89-148.
Goto, s.
1910, On two species of Hydractinia living in symbiosis with a hermit crab,
Jour. Exp, Zool., ~:469-496.
Hnrgitt, c. w.
1906. The organization and early development of tho egg of Clava leptostyla.
Biol, Bull., 10:207-232.
c,
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I
SPECIAL TOPICS - SYNCHRONOUS SPAVTNING AND ITS RELATION TO LIGHT
Baker, E, G. S,
1936. Fhotoperiodicity in tho spawning reaction of Ponnaria tiarc1la. Proc.
Indiana Acad. Sci. ~:251-252.
Ballard, VT. Vl.
1942. The mechanism for synchronous spawning in Hydractinia and Pennaria.
Biol. Bull., 82:329-339.
Brooks, VT, K. (listed above)
Hargitt, c. w. (listed avovo, 1900 and 1904)
Hargitt, G. T. (listed above, 1909)
(listed above, 1910)
Mayor, A. G.
l~urbach, L.
1895. Prolimin.:::try note on the life history of Gonionemus. Jour. Morph.,
ll: 493-496.
Ri ttonhouse, ~.
1910, The embryology of Stomotoca apicata, Jour. Exp. Zool., 9:333-346.
1
-6Sigerfoos, C. P.
1899. A new hydroid from Long Island Sound. An. Nat. , 21:801-807.
Smallr10od , w. J,. (listed above, 1909)
Torrey, H, :a.
1907. Biological studies on Corymorpha. II. The development of c. palma
from the egg, Univ, Cal. Publ., }:253-298,
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D. S!'ECIAL TOPICS - ISOLATION OF BLASTO!.!ERES
Beckwith, Cora J. (listed above)
Hargitt, c. w.
1900. (listed above)
1911. Some problems of Coelenterate ontogeny,
I
t
)
1
Jour. Morph., 22:493-550.
I
!.faas,
Experimentel1o Beitr~ge zur Entwicklungsgoschichte der Medusen. Zeit.
f, wiss. Zool., 82:601-610,
Teissier, G. (listed above)-Torrey, H. B, (listed above, 1907)
Zoju, R.
1895. Sullo sviluppo doi blastomeri isolata dallo unovu di alcunc medusa.
Arch, f. Entwicklungsmochnnik, !:578-595. and ~:1-37.
1895.
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E. SPECIAL TOPICS - REGENERATION AND AXIAL GRADIENTS;
1. REGEN'EK.TION FROM"""CE'LLULKR AGGREGATIONS
Beadle, L, G, and Booth, F, A.
1938. The reorg?~ization of tissue a~sses of Cordylophora lacustris and tho
effect of oral cone grafts, with supplementary observations on Obelia
gelatinosa. Jour. Exp. Biol,, !2:303-326.
Child, C. M,
1928. Axial development in aggregctions of dissociated colls from Corymorpha
Physiol. Zool,, 1:419-461,
Do!Aorgan 1 VT, und Drew, G, H,1914. A study of tho restitution masses formed by dissociated cells of tho
hydroids Antennularia ramosa end A, antennina. Jour, lhlrine Biol.
Assoc. of Plymouth, 10:
~yn~ B.
(listed ~bove)
-Harbitt, C, VT,
1915. Regenerative potencies of dissociated cells of Hydromedusne, Biol.
Bull,, 28:370-384.
Wilson, H. V,
-1911. On the behavior of the dissociated cells in hydroids, Alcyonaria and
Asterias. Jour, Exp. Zool., 11:281-338.
2~
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i
[
IDX;ENER..\TION FRO!A NON-DISSOCIATED CELLS
Barth, L, G,
1938. Oxygen us a controlling f~ctor in tho regeneration of Tubularia,
Physiol. Zool,, 11:179-186,
.
1938. ~unntit~tive studies of the factors governing the rnte of regeneration of Tubularia, Biol, Bull,, ~:155-177.
194o. The relation between oxygen consunption and rate of regeneration,
Biol. Bull,, 78:366-374.
1940. The process or-regeneration in hydroids, Biol. Rev., !2:405-420,
Child, C. M.
1941, Patterns and problens of development, u. of Chicago Pross. (Contains
references to all Child's pnpers)
Gilchrist, F. G, .
1937. The hydroid polyp Corymorpha palma as gestalt and as history. Am,
Nat,, 71:382-403.
I
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li
-7Goldin, A.
!
1942. Factors influencing regeneration and polarity determination in Tubularia crocea. Biol. Bull., 82:24}-254.
1942. A quantitative study of tne interrelati onship of oxygen and hydrogen
ion concentra t ion in influencing Tubularia regeneration.
f
Biol. Bull.,
82: 3140-346.
Goldin, A. and L. G. Barth
1941, Regeneration of coenosnrc fragments removed from the stem of Tubularia
crocen. Biol. Bull., 81:177-189 .
Haz en , Annah
1902, Regeneration in Hydrnctinia and Podocoryne, Am. Nat,, ~:19 3-200 .
Hnrgi t t, C, VT,
1899. Experimental studies upon hudromodusne , Biol. Bull,, 1=35-51.
Lund, E, J,
1921. Experimental control of organic pola rity by tho el ec t ric current. I,
Effects of the electric curren t on r egenerating int ernode s of Obelia
comnissurc.lis, Jour, Exp, Zool,, ~:471-493.
1923. III. Normal and experimental delay 1n the initiation of polyp formation in Obelia internodes, Ibid,, lL:69-88,
1924, IV. The quantitative relations between current density, ori ent ntion
and inhibition of regeneration, Ibid,, ~:357-380.
Miller, J. A.
1937. Some effects of oxygen on polarity in Tubulnrin crocea, Biol, Bull,,
I
,
I
i
I
73:369.
1942. SOme effects of covering the perisarc upon Tubularinn regeneration,
Biol. Bull,, [1:416-427.
Moog, Florence
1941, The influence of temperature on reconstitution in Tubularin. Biol,
Bull., 81:300-301.
Moog, Florence nnd SOl Spiegelman
1942, Effects of some respiratory inhibitors on respiration and reconstitution in Tubuln.rin, Proc, Soc, Exp, Biol, and Mod,, ~:392-395.
Moore, J, A,
1939. The role of temperature in hydrnnth formation in Tubularia, Biol,
Bull,, ~:104-10 7.
!~organ, T. H.
1906. Hydranth formation and polarity in Tubularia, Ibid,, }:501-516.
1908, Some further records concerning the physiology of regeneration in
Tubularia, Biol. Bull,, 14:149-162,
1909. The dynamic factor in regeneration, Ibid,, 16:265-276,
Morse, M.
1909. The autotomy of the hydranth of Tubularia, Ibid,, 16:172-182,
Muller, H, c.
1913. Regeneration der Gonophoren bei den Hydroiden und anschliessende
biologischo Bcobachtungen, I, Athecates, Archiv, f, Entwicklungsmcchanik, 37:319-419. II, Thecatcs, Ibid., ~:288-331.
Peebles, Florence
1900. Experiments in regeneration and in grafting of Hydrozoa, Ibid,,
10:435-488,.
Rose, S. M, and Florence C. Rose
1941. The role of a cut surface in Tubuln.ria regeneration,
14:328-343.
I
I
Physiol.
Zool.,
Torrey, H, B-.1910, Biological studies on Corymorpha, IV, Budding and fission in heteromorphic pieces and the control of polarity, Biol. Bull., ~:280-301,
Zwilling, E.
1939. The effect of r emoval of perisnrc on regcnoro.tion in Tubulo.rin crocoo.,
Biol, Bull., 76:90-103.
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-8PAPERS
l
SCYPROZOA
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Friedomann, o.
1902. Untersuchur~en ubor die postembryonnlc Entwicklung von Aurelin
Aurita, Zeit. f. wiss. Zool,, 71:227-267.
~
Ibrgi tt, c. vr. ond Har&itt, G. T.
1910. Studies in the development of ScYPhomedusne, Jour, Morp., 21:217-262.
Hcin, VT.
1900. Un tcrsuchungcn uber die Fn twick1ung von Aurolia nuri tn. Zeit, f,
wiss. Zool,, §I:4ol-438.
Korschelt ~ E.
1936. Vcrt;loichvnU.c :r<illtwicklW1(:; SC.J::>chichto· d cr Tiero, ~ena.
!4ac:Bride, E, Tl,
1914, Text-book of E:r:lbryo1ogy. I, Invcrt ebrntc., I.incmillan,
Percival, E,
1923. On the strobilization of Aurelia, Quc.rt, Jour, Micr. Sci. • .§1:85-100.
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DEVELOPllENTAL HISTORY OF M:n.ANOPHORES IN :BIJIDS
~
!
En.stlick, H. L,
1939. The point of origin of the mo1nnophoros in chide embryos ns shown by
means of limb bud trnnsplunts, J. E. Z,, 82:131-159.
Foulks, J. G-.
1943. J\n ~nalysis of tho source of melnnophoros in regenerating feathers.
Pbysiol, Zool,, 16:351-380,
Hnmilton, H. L,
-l94o. A study of tho physiological properties of mo1nnophores with special
reference to their role in fcnther coloration, Anat, Rec,, ~:525-547.
Ra.'71es, M, E,
1939. The production of Robin pig ment in White Leghorn feathers by grafts
of el:lbryonic Robin tissue, Jour, Genetics, 38:517-532.
194o. '!be pigmor.. t - :ornine p o tency of cnr1y chick bU:istoderms, Proc, Nnt.
Acad. Sci., ·~ 6: 86-94.
1944. Tho migrut~. o~of rnoln.noblc-.sts after ho..tching into pigment-free skin
grafts of tlv; c or.n:10:1 :'owl. Physiol. Zool., U:l67-183.
An ex:poril::~.-nt~.l s l".udy on tho origin of molnnophores in birds, Physiol,
Zool, • lll: 4?;- G6 .
Watterson, R. L,
1942. Tho rnorphot:; '-:lnos i s of dnrrn feathers with special ref er ence to tho dovc1opMcmtal h is:or;t of nelnnophorcs. Physiol. Zool., !2.:234--259.
Willi or, :B, H.
1941, li.n unnlysi s of f co.th rlr cvlor pnttcrn produc ed by gro.fting melanophores
during Cf.lbryonic dev c bpr.1cnt, AM. Nat,, 12.:136-146.
Vlillier, :B, H, and R:'Lwles, !~ . E.
194o. Th .; control oi' fc<.'.. t hJ.~· c olor pa tt er n by moln:1ophnrcs gro.ftod from ono
eobryo to .-.mo ·~;:n r of c diff <J rcn t bruod of fowl, Physi ol, Zool.,
ll: 177-199 .
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T e tbrce for; s to h; st '!.d:!.ed r.re H';~l.,ohi.e s ( E'..l'Pll'!~"' t :s)
hc:za:~on '..S ,
-
•:m 4. Sa~1el l£\l'H\ •
:'G 1 'CiS
C lt 1'3S of "- ·vc.. . c~ d sta_~es \'.iJ.:. 1:e :•eun:•e ' .
f1
r:,•e::>are yo;:.!' o···r.. Ctlt;."''C<; :''0C Gd ::'S follo•··s~ :r·-,rd '
rnd fcr:nles, ··:il .. r.'"'·."n iP!1Gd.i2.tel·.r rfte~' ,0i. <': re:1
cr lc ~r eous tu'Les . R0!'1ove sever:'.. ... :1nd n l r.ce tbcm ~n
o .c
~orp ner dish to een t" e se xe s se~~r~~e) .
t'·e;r t:ave s:;n··.rne-.::. . f_et s;)e-·:·: stan(\ for nhout
-~:20T') S of snel'm sus rens:.on to a dis:l of e · ·~s .
hooi es ancf cl~9vn-e r".&·- be e:..1sil" stn . icc') • T•:c "·lf'.st 110. st·, · ~s
re'1ched ..,fter 5 - 6 !lours , r-;nstn.:ln+-~on ::>ft·'r 8 - 12 :1o1.,rs ; tle t r"c 10 :.c
st"'~~e lasts fro!:, 20 :1otn•s to t·:o vJGe\:s . -r· c troc'·"~;• 1 10"S ·.r· e 1er.t fc
st 1dy ··r-en 2 - 5 day s old . s~bell~'.. ricl. .i:lf;,- 1 c tl' •r.t.: in t 1 --~ <" [tlllP. \.'f':: .
Fe"tilizati on of :i-Tel'eis l'C.S t.' Se:l sb'.died ~'l' 'l ·-.:..·;:: ~J.ons ln ..~ . :!el"l. d .
0
I.
~
. P~~ '
... .... ''e"
. .....,
Tl·,e
Troc}'oph .;r c C'L
h'"~
\.
.
rn:c t i' OC] 10';"\!l o :.'e is a typlC ~~ 1 Ann~J.id troc·~ Y''-ol'\) . Con:..~nlt tl'O
e:;~collent fir;s . in ra tsc :Jo~c (t RG) 1. .~1d. S> c ''l' Cr (t\1) . ·p~~8 lnrvs c ~l'
positive p'11 otot'l.XiS etn.d. c:a t~1el' 3. t t'~ c \'l in clo\'! Si ll,(' ot t~1e (.j ~\ ' • t: Dl.'...' t
tJ~ochophoTos , 3 - 5 cb.:rs old , on u slide n '!. :.ell :'. f8'".' ::..'iJJ'Oll.n of J ~1s
paper 'have b0c·n plac..::d. t.o (,mtnn~~;lc t~101t ancl hol · t· en ;tu\.-t .
!f1r cotics , c . ~~ · n fm: dro;s of ~llo!'ctw.. J or of l :-SO~ , c.J.'C no i: vor;•
effective but may . \.,; trio~.J. . Vi tt1J stain).n:· obscm•.:.•f\ rr th r t>:.m
cl:?.rifi c s t1 Jc 's trnc:hn·e s . 'r ' :e l~·:· v:->c c.1~0 trr.ns:9oi'Cnt , 1.nd pr.1l)Cl'
Rdjustmc:nt of t.J.O i11-w'linn t:i on :Yy YJ10VH1._:;; t lJc r~::.7'r or nnc.1_ t l10 _().: '0
conocnscr 'i':\.:1 ·)rinr- out c.Jl s tructn:c'Os . stnd·· ;-nJ.:.!n ls J.n l"'tv 1 't:"l
2-nd in poh~:r· viC'.'':J ( bot ··, f·c01; ·~.nit1r·l :-.r~(~ :":eo' vce-ctaJ. ;•o le) . 'T'lc
apical tnft and t: c annl vc;r.:Jclt; 2.1' ·J )_8llC~mo.r l~s for t:1o poles , t.1o
mout!l is on t>e V(;ntr:tl :.:.itl_o ; t···o C!?t".' ~.f.\ on t l1 C ri 1;'1 t sid,.; . 0 1):.>-..:!'VC
the loco~ otlo r firs1· .
Observe:
1) Sh8.:pC cf t ''c t:roc'lop'!orc .
2)
-~
>)
l' ,... .., -:
!:.!..~
a "'
....
'"
(....
1- ., '.L' i -
~
-·..
~.) C' . ~:.,
)
'oJ
( IJ
..., ..~ 'T~
,,
o·t-.r'' .1 )
•
...;,).;..;.
l'" '1
l0n:::.
eiJ.i[~ ,
•
3 ) ,\ :- ic--· 1 "ll' ::; n , . t;}: icl{E- n Jn:: or o cto c1.;:n"l ·-~t ·i~L... '1.'lit.cl1.
:•ole ; n. n•Jrvc c.._l~ tc :· end ~w·ol'n ') ~.y the! ;r J..: r. Ol'l;.i,t:YJ. o~' t::c
.,c,.,r:1Jl,C. J . :-~~~~~J.i8 .
4 ) l'rc !r::;t-o t: ·rL~!, , :.'no,,., to~·i .. l 1lnlld nf l ·~_:c r.""h· . • i . tl
older t ro c ···n,:1o . . · l.)f; , t'!!O r o\'iS !.' c 1J.l.:. \:~ :! :' I : L·o:"ult:l ; a
row of sY.o!'t cJ.J.iv c~n tcrJ..or to t.•c 1::£'[';...: c :. J.i,1 . 'l'l o
_01 otvt:coc'1 is tl--c 1110'-t chrtrn t0rl:)tie ~trueturo c f t'1.c
J.;u'va , DY1c:. [';::t " ·2 1.t l t:G nttr.JC . It:.::. 2- l\J c:,r a rnt...:1ior to
t>c nout> (pr oo l': 1) . :rt con~nst~: of :. few :.r:•.,.,J p . . 0t (')troch cel l::.; v!hich1'lC~Ol'1C p:l.{::; . 1cntod in older 8t~trrl;::; ,
- 2 -
:; ) ·-... ·. ,_ e ~
. . '1 1' '~~..~·-".... _._v....,., • c·-·
· ) , a c .;.~.. r c · ......."-, r
....,
•.• e ....~.~ a ~v r o .....
:-:idd~.e cf · t:w osttroc: .. nJ. ~ .. el::i<J:-.acre .
be.r.d of ci ~.in
t 1 ~- s:-, :'. ia t"'q p·r -:oy~ ~~ t::e 1::id ventra~ 1 .:.ncl co ~ .e c ~- hlf
:::0 ·.-ch ar.C: a 1t.:.~:
·::-.L. ·:r<. c e is interestLlf in t.:, ·
.:c.~·}rs t:l') l.i.. '"'C: o.~ c.:LoSUl't.. of t' o ' l • to_porc .
'1."-c •. · ~~
·c.:.e re:iinant o:'
t:~e
opening at
7,
"
blas to.~:- or~;_ the cnu is a H c nf.ary
1. ·-..·Jer end of t_ e riginal ' lc.!' t O.J:)Ol' . sl i
cl.~
One eve on t:"1e !·5 ·J: c s id€ of the pretre>chnl
:iote tlle red e:J ~ p.:.gn~ent .
~- .
:~o~i "'P':turt;.
8) ~"·"0 s ta t0cys ts on the v cn tral s id •
9) Tlle digest_ive tract, con::isting of:
mouth opening,
s tomacJa,.um
oes op\~.asus; e c toden~.al), enlara:ed n t r.mcl-,.
-r=
( entodcrmal) , narro ·: int- s tine ( entodermal exc"pt for t' e
end portion Y!hic~ is ir~0 gino. ted ectodel'm
proctod eum),
and t'1e ~' an o ; ening '.•e'1ind tLe v egete.l f • le . All partn
are bese t ~ith cilia. Feed india ink and otu~y - ~~e mechanism of feod intake .
=
le) T:1.e anal vesic~~ a larc- v~?cuolP. tcd cell at t.1e
end ,--no£ found in other -::.rochophoros •
ll)
p e r~e
'ior
T:1.e cavity bcty;een i:1cv ti ~1 e and outer L.,;Ody ·;all lc- not a
true coelom but a prime.:.."';r •b ody oc:..vit:;[, the pert>i~ting
blastocoel ~.
•
12) Tne larval kidneta .(p,.,ired) are typical protonephridia ith
flame cells; they open norr the anus . They appec.r r..s
slender cords near the st.< to c ysts , extending c-c·-;oen
oesophag-.A.s and anus. '11 :~.oy are best identified in nnimE'.J.s
~it~ the vegetal pole up (consult figs. in Hatschck and
Shearer) .
str'-~~d:.; 'Nill be seen bifurcnting at ·c:~1e
·uppercnd of t . e lr.r val l:·idney .
One of them O".n oe trr·ced
to its insertion at t l1e rpicol plate , the other inserts at
the oesophe.[l;USe Thes e o.:c·e ~-ongit_Ejj..nf?].__n:usclEs .
Ctl'l•r
lonc;i tud :t~e.l mur; cleo extend from tho stomach to 1Jointo
of the -c..r _t:)cr '~-:':'~n h: phero . A s tr0nr.; e; ircule.r mus cl o is ner. r
·'-'1.-.e
~,.,,.:I.Jc.
. ..,.~o-v L' "~h·
t'~c
"J.l,:
l"rV"
C"U<-'C-'1
b•r l'tn
v: .t il!v
- '- - !
.l..
'··o
'
Dv
v J. c ·'·
1.J 4 on of -,:-,
L,_. 10
•
co
<
'-•
U.
contr r-.•:.tJ on vriJ.l be frecluE.mtly ob[Jerved . Nr.-·C.e s.lno cir-~ulrr (s p.linct e.£_) !!_1~scl<:3_ 5.n tl•.e di~e~ti v e -Lrrct .
l.3) :.ms c lcs . Tv.ro fine
.1.
o.-
•
,J
14:) Undtfferentinted octom•n ode.rm c elJ.2 , single or
"-'
in rmall
c,roups , vlill be seen a·~tc.c 1 ·ed ·to t':te stomo.c:1 , to the inner
~Jod.y 'Nall , near Vw ap:t.c:: 1 organ , et c.
15) 'l":Je important cntmres odcri,~al cells
( del~i v:a t i v:;s of 1d :
' '; hicll v:ill. [< f"v-erise to tho 171€lSOdCI'I118.}. struc'(:.ures of tl1e \;orm bod~r e.rc difficu1 t to dis tinguis ..1 . ' t> g j
are sma.ll groups of c ells near the lower end of t l• o )t_ead
kidney .
r.felo!_:~lests )
Dr a.\'/ lateral and pe>lar vie':Ts.
-3-
II .
~'he raetamori-hos is of an An. elid larva into t. se :n.e'1ted worm can
e . t· died best in Nere:ts, 1 to 7 days old . Ne re is _a~ no typicnl
Lro~~o 1 ~ore but an a breviated, telescoped
arval developnent. The
;irst signs of the adult, segmental organization ~\i:-· .dar very early.
:= rc.:-c.re s J. ides as under I. Study larvae from a 11 sides. Consult the
i}fS . in E. B. Wil son {'92) .
A. Tr ochoyhore -like stE~.ge!. 24 hrs. C'iilson, fig. 84)
Observe:
1) The prototroc~, composed of 12 very Jarge ciJ iated cells.
2) The paratroch, near the vegetal ~ole.
3) A pigmented area
at the anal 1-olc:
the anal pigm nt.
4.) The mouth and stomodaeum; the latter is a short ectodermal
invagination. - - - -·-
5) The large maororr.eres have not yet differentiated 1.nto the
entodcrmal parts of th13 Jn t os tine; their cell boundaries may
be seen . No anus is formed as yet .
6) Several
frontal bodies 11 near tho UPI>er end .
like structures of unknown function.
11
Circular disc-
7) Two pairs of sc ta sacs , s phericnl s true tur es L: the
os trochal
hemisphere . Tho s:.:;-tae { = chaetae) will be differentiated
inside of them . 'Tbesc sacs are the first ind i cation of tho
first two s cr:ments of tbo worm.
8) Obs c rvG thE; trochophoro in locomotion.
B. Advanced trochophoro, 2 days old c~·,· ilson , figs . 85-89)
Ioontify all structur0s found in A.
In addition observe:
1) EY,e spots •
2 ) A third f•a ir of S c ta SHCS has a1 _~,.• oarEJd 'behind the first ·two
All: thrC'C'"8:r0lined ur-in a rO'H indicating tho first 3 segments . Setae c an be seen in the {>rec ess of formrtion , inside
of those ss~s_; jn sJ.ightly older stages, th ey v1nJ. i)c se en
projecting from tho sacs .
3) 1-' igment a1-•1- 0 ars in the urototroc.h ccJ 1 .
c.
Metamor 1 hosis . 2~ -3 dnys l~llson , figs. 90-91 .)
Notice the change of shape and the gro.dual demarcation of th, first
3' sogments.
-4Observe the follcmin[ no :v
13atur s:
1) Sense h.airs at the a ~cal pole.
:2) Additional ~ spots; nu.•1bE:r variable.
3) ..'ie sotrochal ciliary bands at the boundaries of th" segments.
4) Pa ra};odia with parapodial cirrhi appearing on segments
2 and 3.
5) Tentacular cirrhi on the head.
6) Stomadacum, stomach, int€stine become distinct.
This stage comb ine s larval (trocho};-horc) and adult strnct1.rcs
in a peculiar way.
D. Late Stages of ::etamoryhosis.
(1.'dlson. fig. 92
4 days and older.
~he head has lost its balloon shapo and devolO.i:-S the a~ 1 ondagos
charact eri stic for tho adult. Locomotion is still ·~y means of cilia.
The end of metamorr;hos is is 111arked by t;"~e shedding of t~1o prototroch
2 nd t!'.e mosotrochs, and the change from swinuning to crawling.
Obs crvo:
1) Frontal antennae, tentacular cirrhi, palpi, all on the
h~ad.
2) ~ cirrhi.
3) Two la~ and scvcr1:1. l small eyes.
4) Jaws inside of tb.e oeosphagus; they · can be protruded.
anims.l is carnivorous and rapacious.
'l,ho
5) Study the finer structure of tho setae, and their motility.
(muscles at t.'!eir bases).
III.
Tho 'I'r?EhOlJhorc of Sabellaria.
This larva shows very long bristles VJl:.ich hove pr oba" ly both a
suspensory r..nd. E:. protective function. Study trochol- ~·10rcs, 2 days and
older (consult D.P. V1ilson '29, ~nd Novikoff '38).
Obs <Jrve:
1) Stiff apical cilia in t:-:..e
}!]
ace of tho apical tuft.
2) The prototroch consists of 3 rows of cilia, and shows a gap
on the dorsa! s j_de.
3) The neurotroch in tho midvcntre.l line.
- 54) On0 Gye on
t ru ).eft s: do .
5) l"he hOOd 1 ~ fold
~:or·
s, s ots d 'c lo.:: 1nt r .
OVC rhL!1Ci~[' t£1 · l OUt l o
C) Very long b r istL. s d e' -.1 01
in s ..:tr, sacf' . St :tdy tLt ir f:tnc
structure . T_~DY D.f•!IG ar on0 _ r ir ~.ftcr fm oth: • , '"'"" n t tally
10 p~.irs . ThEly ;-;ill ·~ 0 r p lc. ccd ct mctamor h osis by
ordincry sv t a El .
IV .
Grstrulntinn Lnd Forrr:r.tion of the
rl'ro~roro i n Hydroidos-.- :r r cpa rc vour own C"ltur:s (s oc p . 1) . Gustrulat.ior:. by inv~r·i­
nrtio~ occurs c~proximatoly 7 - 10 hrs . rftcr fertilization . Consult
the figs • in Shvaror ( ' 11) ~nd Ho. ts chck ( ' 86 ) •
-Qrv · .
II
.... .
190
~a- ~-Y c~s ve~ opl.r.ent
:>c . 9
of Are i col a .
~ou=~ I
.. rc':1i v .
Fraipont , J .
1887
Le Gem:·e .~. olygordius . Fau!1a u . Fl ra Cles
Golfe s von .Tes.pal , Vol. · XIV .
:1atsc·-_ek , 3 .
1886
Ent·· icl:l·lng do:- Troc hophor e von Eup oma tu:5 .
Arbeit . a.d . Zool. Inst . ~ ien, VI . ( ~droi 4 os)
1928
Ent·· icklunt; der Larvr. ls egncnte · e i den
Anne lide.:1 .
Zeitachr • . or ph . Oel: . ... d . 10
1922
On t~e r earing of sexually mature
e ggs. Am. Nat . vol . 56 •
1902
Differentiation .'!i thou t olea vase i n the egg
of the anneli d Cheetopt c ru~. Roux Arcbiv' f • .
Entw. Bd . 14 .
1006
Observati ono and exve:riments concerning the
eleme ntary phenom. of embryonic dev ~l o ~nent
in Chaetop t c rua (Forms. tive ~ tuff) Jour . Exp .
Zool. III .
1909
Pol arity and bil a t e r lity of t h o Anne li d egg .
B i o 1 • Bu lJ • XV I •
Mead, A. D.
1897
The e ar l y development of marine 8.nnoJ ids . J
•
Morph . vol. 13.
Newby, W•. 1fi .
1946
The embr-.ro logy of the Eohiuroi.d ··orm. Ur c c hi s
caupo. J.:,em . Amer. P~'lilos. S~c. vo l . l 6 ]_...,219.
Novil:off, A.B.
1938
Embr:ronic determinatiQO in the J.nnelid,
Sa'bellaria vul gari s. Bitl. Bul l. vol. 74
Shearer, C ~
1911
Development and structure of the troc h , phore
of H~rdroi de s, (,uart. J. :M icro. Sci . vol. 56 .
Wilson, D. P.
1929
Larvae of Bri tish Sabellarie.ns.
Bi el. As s oz. N. 5. 16.
' 1 ils on, E , B.
1892
Cell lineage of Ne reis. Journ. of
Wil son, E. ·B .
1898
Considerations of cell lineage and anceBtral
reminisc enees. Ann N. Y. Ac ad . Sci. vol. 11
Wistingh ausen,
c. A. von
1891
Unt e rsuc~'lungen
Just , :: . E .
. Lil l ie , F . R .
dumerilii.
Bd. 10 .
oruis from .
J. l:e.r ine
M or ~h .
v.6
Uber die En t·..ric kl . von Ner eia
Mitt. a . d. Zoql. Stat. Neapel .
1904
'f!u rrnkol: f , ·::urmrumpf und Trochophora . Zool.
Anz. Bu . 28 .
H04
B eitraege zur prakt i sohen P.nalysc der 1-' olygordius-Ent·:•i oklung , etc. Roux Arc hi v. Bd . 18.
Zur Kopffrage der Anneliden . Ve rh . d . Deutschm
Zool.Gea. (His figs . are re produced in McBrt.cie)
1905
ARBACIA PUNCTtDW\TA
Fertilization
Gametes of Arbacia are best obtained b' cuttinG ar und the
peristor.:e ~ on oral surface) arrl renoving Ari totle ' s lantern, tnkine
care no~ to injure the gonads. T e pe~ivisceral fluid is then drained
from the body and the animal, aboral surface do·n, placed to shed on
a Syracuse 1"atch glass slightly moistened '''i th sea ..ater .
Vash the
hands and scissors v~th running fresh ~ater after each animal is
opened to avoid contamination of one sex •ith gM.. tes or body fluids
of the other . There is no convenient nethod of ascertninin the
sex of sea urchins without cutting into their perivisceral caviti~q,
but ·vrhen this is done , the ripe ovaries v:ill be observed to be deep
reddish brov111 in color , and the ripe testes a gra~·ish v1hi te, \Vi th
flecks of brovrn. If the eggs are shed through the gonoporos of the
female into the Syracuse dish , they should be removed to 200 cc. of
sea water in a finger bowl within ten minutes , to nvoid the injurious effects of crov.rding . Tho sperm are best kept " drytt ju<.'t ns
they exude from the testes . A drop of " dry"· sperm may be diluted
with 10 cc~ sea v:ater in a v;atch glass just before insomina~ion .
Do not use sperm that have boon diluted more thnn 20 minutes and
avoid too high a sperm concentration . The latter leads to polyspermr
and abnormal cl eavage . 1\IJO drops of diluted sperm should be sufficient for a finger bo !fl of eggs . Stir the egg dish imModia tely after
adding sperm for insemination .
If shedding of the animals does not take place v•ithin ton
minutes after removal of the lantern , rnake a second cut around the
shell near the equator , just above the oral ends of the ovaries or
testes. If , after this second cut , gametes do not stream from the
gonopores , ovaries may be r emoved with curved forceps to 200 cc . of
sea water in a finger bov•l and the ripe eggs vri.ll be extruded
through the ruptured ovarian walls . The egg suspension is then
filtered through cheese cloth (v.rhich has been soaked in sea water)
to remove ovarian debris , and after the eggs settle, the sea vrater
is t~ice (at least) decanted and replaced ~ith fresh sea water to
remove tissue extracts . Vlashing ot' the eggs shed after cutting
around the peristome may nlso be necessary if an ovary has been
i n jured or tho eggs oth8rwise cont aminated vrith perivisceral fluid.
Test es may be simi l arly removed to a dry vatch glass with forceps, .
and exu-ded " dry" sperm used as indicated above .
A good batch of eggs from a ripe female should show unifonnit~
of size , perfectly spherical form , and complete absence of immature
eggs ( in the germinal ves i cl e stage ). The egg of Arbacia is shed
in the fully mature stat e , both polar bodies having been given off
in t he ovary .
Very rarely, t h e pol ar bodies remain on the egg
surface . Usually t hey are not present , and the onl y good index of
polarity of the egg is the funnel- shaped space in the jelly which
marks the previous point of attachment of the egg to the ovarian
wall.
The unfer t i l ized eggs should be examined in a drop of sea
wa t er under a cov er glass and the f ol l owing parts noted: jelly
l ayer ( rendered r eadi l y v isible wi t h a littl e J anus green soluti on or by a suspension of Chinese ink )~ vitel l ine mernbrane (very
Arbac ia - 2nature egg nucleus (a small, clear sphere); protoplasmic
:,,··:u ~ule s of t 'IO ki ':lcls, ~ 1
red pig;ment granules o nt ning echinocnr ,.x.,. ,
·~· ~aller, colorless yolk granules .
o other types of granules, ioe.
~b8 ~itochondria and the oil spheres
represent but usually disting,...:::. stabl~ only after stratification by centrifuging.
:.L·.• _, :
J.nse .ina te some of the eggs as indicated above nd e. or.1i 110 L~c. o(t,_,nder low power. Sperm penetration occurs too rapidly to be
easily ! e ~ected . ITithin a fevr seconds after insemination the cortic~l
reoro~'"'ss of the egg begin. The vitelline membrane elevates rnpid1y
from t.:te Ggg surface, leavihg a perivitelline space. T is mem rano
thickens and hardens during the next five minutes and after altcratirn
~.s called the fertilization membrane. At the protoplasmic er,g surf cc
is at first slightly disru~d by the elevation of the vitellin~
1 which
r!.lembrane) a new, clear layer is secreted - the hyaline plasna mertbr n ,
··1hich is presumably a calcium-proteinate acting as a cer.tent to hold t.h€'
biastomeres together after cleavage. It disappears in calcium-free
sea water. After insemination, the jelly layer can often be clearly
made out by the ting of sperm entrapped near its surface. l\~oser ( 193 )
has correlated the elevation of the vitelline membrane' ith the brcnkdovm of certain cortical granules. These granules are embedded in the
cortex and are not easily displaced by centri~uging as are the granules
of the underlying fluid endoplasm. In the centrifuged eggs those
cortical granules show up clearly at the hyaline zone.
i ' l ~·.ely
Examine eges centrifuged in an electric centrifuge at 6,000 to
10,000 times gravity and identify the foll~'in~ layers in order from
centripetal to centrifugal pole: (1) oil cap, (2) hyaline zone containing nucleus, with cortical granules visible at periphery in optical
section, (3) "Fifth layer", or so-called "mitochondria, (4) zone of
yolk spheres, (5) zone of pigment granules.
Mount centrifuged unfertilized eggs under a coverglass on a slide,
and focus on cortical granules at hyaline zone under high power. Then
carefully add. a drop of dilute sperm at the edge of the coverglass.
As the spermatozoon penetrates the egg there is a wave of breakdown of
the cortical granules, which is followed by rapid membrane elevation.
The cortical responses of the normal and centrifuged eggs to insemination may be slowed down by decreasing tho t emperature . This can
be done hy placing small dishes of eggs and sperm in an icc bath and
cxaming them on a chilled slide after insemination.
It is also of interest to examine fertilized and unfertilized eggs
by dark-field illumination (even a dark-field stop disc placed in an
ordinary condenser is often effective) and note the chango in the egg
cortex upon insemination. This disappearance at fertilization of a
cortical layer which is bright under dark-field illumination was
first described by RunnstrBm (1928).
Make sketches of all distinctive features of tho Arbacia egg
mentioned above.
In&uguraion of
Arbacia -3Cleav_~
The time between fertilization and first clcavc gc va ries
slightly for different lots of eggs at a given temperature. Some
deta ils of the mitotic process (including spern a ter, diaster a n , '
spindle) can be observed in theliving eggs, particularly if they are
slightly flattened by compression under a covergla ss. Reme .ber that
~ggs under a coverglass develop normally for only a short time a nd
under compression this period may be markedly decreased.
One of the first internal phenomena clearly visible is th
formation of the sperm aster, which appears as a round, somewhat ole r
region at the time the pronuclei come into contact. The sperm aster
then elongates, to give the "streak" stage, with a some hat cl ea r
crescentric streak extending across most of the diameter of the e r,g .
The streak then fades, and is relJlaced by tvo clear regions, mark i r.
the two asters of the cleavage spindle . ~he following time schedule
applies to 50 per cent of the e~gs at 20 C. (Fry, 1936): Round spo1n
aster, 11 min .; "streak" stage, 30 min .; fully fonnod dia s ter, 4.5-50
min .; first cleavage ,...-:::----:_-,- 65 min ., second cleavage, 106 min . ;
third cleavage, 145 min . It is important to note thgt first cleavage
of some eggs will occur as early as 56 minutes at 20 c., and at 45
to 47 minutes at 22° c. Observe and sketch cleavages through th 8
cell stage, keeping a time-schedule of events at the temperature of
the laboratory. Note the behavior of the hyaline plasma membrane
during the cleavage process, the relation of tho first three cleavage
planes to each other and to the egg polarity, and from time to time,
examine a fresh sample of eggs under compression to observe such
internal phenomena of mitosis as are visible in these living eggs.
References
Boveri,
Th., 190?. Die Entwicklung disper:q1er Seeigel-Eies. Ein
zur Befruchtungslehre und zur Theorie des Kerns. Jenaische
Zei tschr., ~
Be~trag
Fry, H. 1936. Studies of the mitotic figure. V. The time schedule
of mitotic changes in developing Arbacia eggs. Bioi. Bull.,~ 89 .
Harvey·, E. B., 1940.
A comparison of the development of nucleate
and non-nucleate eggs of Arbacia punctulata •. Biol. Bull., ?9.
Just, E. E., 1939. Basic methods for experiments on eggs of marine
animals. Blakiston, Philadelphia •.
tiorgan, T. H. , and Lyon, E. P. 1907. The relation of the substances
of the egg, separated by a strong centrifugal . forve, to the location
of the embryo. Arch. f. Entw. , 241
Moser , F. , 1939. Studies on a cortical layer response to stimulating agents in the Arbacia egg. I, II. Jour. Exptl. Zool., 80: 423,
447. III, IV, 1940, Biol. Bull., 78.
Runnstrom, J., 1928. Die Veranderung der Plasmakolloide boi .dor
Entwicklungserregung des Seeigeleis. Protoplasma, iL 388.
a1
n
J
or
ions into
(see 1935, p.
reepec 1 vely~
into
o ro a
pectivoly. • e
ce e those of' th
tho clea-vs E'
1n normal d~vclop~ent
1c nce" 9 Drit~c ),
de tineti to bec-c
of th~ pluteuso
copied "' om tbiE
see Hoerotadiutr
:futur
.&.CJim the
keleton
~. p. ~59, and the chart
figure~
Concerning
c eavage pat
r
g28, lB39.
3, Blastula. P. ent:ral blastocof'le appear3 t erly.
>laatu:a 1 e ·. pheric
at :first ar.d beco:. es cJ.liated. It .t.latt ne aoon o th. · •get~l pole,
d at
the Eam~ time the animal Jole develope long, etiff cili
tle ~1 tuf •
'!he ... light thit!ker.1Jl!: cf the blasto·•ocl
all at th~ apl.o.ll end is called the
apicc..!_ 2..;_f5.8Do Primary me. eenchyme c lla. deri vnti ve of the micl·omere
auder
into the blastocoele. 'I'h~ embryo hstchaa in thE: lo t blact l.e stag •
GastruJ.ation.
The fht"tened vegeto .• area begins to in ag1 teo 'I'he ar
e teron growp first straight upwardp towards the unimul pole
The g ·~trula
asaumee a cc.ne or prieru shape. '!be priLlary m1Jsenchyme celJ
r.rauge thensel e
in a ring arounQ the archentercn n~ar the blastopc·o. They form two dense egg
gatione on two opposite sides of th~ bla~tocoete. The ske e on akes it first
appe~rance i.n t/l( fonn of amall tr1.rad1ete eplcu.l.ea in the cent.
of these
c.lllstorso Obaer € those spicul a. 'Ihe unte:r1or b.lil1d en.i of the intestine
beode to vne side; the ra· 1a Ly s~1metric l larva thul cq~ir a ilat ral e~~
metry. NoUce
e distribut.on ot tl'Je pigment granulev. Sc .. ond y m enchytle
eel owe .e1· 01 .. !.0711 the d:\at, blind end ot the arc:hcnt
, inally~ thi
eDd meets a ah'lllo in·:a.gination of the .ctode1·m, t o
(l um
and t e mouth
breal thro 1gl:l, ~kf.Dllhile tae oalci1.un spicules have g..,.ow c:u. 1·1to long akelet l
rods n the c·our e r;f a ~e, hou~a• Qbe rve as r1 y deto1l of the :t'ormntion of
the kelet;m a- pot~e1b. e~ rot1.<·e th
k l ton &c; rc. :iJ: 1 ~ mob e cells bich
E::Ie ::.1ued up in ro11CJ (v n UbL,.hr l«a'W),
4,
l.
r
is
.>t~·oJ
y 'ar aJ.;
1-:.
.~
devlc sen! adaptttlJn
It hne not!"\ •
CJkeletr. n.
Stu~y 'Jld d.r w dif'~erent stnges (f the tr 10 ti0.1
the pr1
0< :!.....e El::~.:'!Jil__ !~-~r,heped l'lt gtetl"'..lln into tha p.ltl'tet•.G.
Ttlt. ~:~n1;na1 ol::, tec::>gniZfi'Jlo by ita aplt:ol tu 1 growa o• ··to f•Jl'tll tb
ohorter a~·a:l.~ or ·m .ero :.aterc1l arnl, The ~ipJ.c l tuft !.E> •Joon l.o s~o.
h rtly nfter
these a:.. s ,,H e bef?-In t.> t~ro. >Ut~ t w anal C).:nllS gi"O• ont. 0!1 tue an l. o de, ne r
the p">i:tt.s wbf.::..•e -::·.-1e sl.:eletr.:l epic· , et~ ·r1g1nate~. s·, ely th"! g10 th c1' the eizelcton
JlO t.hc gradLL'il eJ.•~ r..orut;.on of t!,e skeletal patt 1T.~
•' D'7 th , IDC6< derm C(; ..l.l.S 1 ic b
;\-,nn the ::;kelcton (ukeletoblasts,~ and othc: memmchy;nt.. cell in the COihY hkh
1:·~,: rt•)~ \rl:,J.<.\t:u ! ' ~·~,_clc":or. fo"~:"IOatior.t. '!'he ma:l.n cavi · o f t e lnr a ie the pe
u•.~·ting 'blastccoc_.<: enJ lOt a t .. t;,e coelom 'whic)l. by d~J'in.lt onJ ia lined VJ1.tb
•• e~odei 1&1 ep1 thelium o Gonf"u. . H erll adiua, 1935 0 fi ~ 2, p. 1: h9 ( (!l.d t.h chal',
ttl.ch if~ a copy of tllis .. igure; foi gcntrnl crien atjon o1' tho lxial l' lations on.d
ccmpa.!'i.son of gastrula and ':lluteua. 'J'he origiue-~ an.. lE l .. vf~get~ll
J.s of the egr.,
< bl stul~l ... u no L cur"Ved lln,.~ • ~ounec t:in.r; t.be • ppe .. p rt ,r th oral lobe wJ. th
~1c or·
field ia venc;l'8.l and th~ ap ~ o'f' th..l p.lut u ie d1>r·aal.
1
ott
It the an1roal opened 1 a e male r the teste a wilJ. be ~hi te or 1 vory. Since
J.t 1.a important to use a fresh sperm euspenaion, thJ.B anLDl 1 mny be placed in
a iry fingerboVJl un-t:~l t.he eggs ere ready for fertilizat ... ono Then a singlt
testis is :;.~emovedp rl.nsed in clean sea-water~ and 9 :~!..:!.'-}.}. _:eiecc from the blunt
eni cut off and placed jn 200 ceo ot sea-m.ltero Two or threE: pipettes of thia
su:lpena1on should bE. c.Jed to a l ?50 cc. finger bowl of egg , with an immediate
but not "ioler.t rote.ti Pl Ino,cment to .ensure complete mix1nr;. The optimum
period ror fcrt1li U3 tion ifJ after t,nf! brE:t .led own of the f enniuaJ. vosicJ.o and
\..efore the lst oolaz· body baa been e:.tti·uded. I .. is theJ efo.C'e convenient to
ln~eninate wt~n the.dlAtll end ot the f1rot maturatiou {pi~dle beRins to protrude ahov~ the previouel)' SUlooth au race of the o""cyte in ti .air pcrccnt8f'e
uf the eggs ahowing genni nal v .sicl.e break.dolln. Egge 11 seminated 11 the atne:e
of t'1e intact germinal v s1cl'9 axe noi:l-fertil.izoblen E ·en tho\.gll they may
~J.e ate a te:rtilizat1o:n merobraue they do not deve J)
fur h r
Cle
The blo ... tomer
.eriv1tell1n
~pee
r A .. t
E
s
ide
than in t:J.e
.rb .3ie egg
~dice
nd the fer 111z
ebsense of tl:e fe
ple ely. Be
se
f the 2,1 ing astera '1,.
form is usuaJ.ly not unje":'t ke
speci l l1terest
g~
h
met
l
1 •
1 t'i oence
- ter Stageu
l.a
Tb raise Asteri
onic at gcs. it ia
sea- ater 1n the cu '· ;! c ish s at b ut ho..... -hOUl' int
t.o elimi ate the e:we~s .. perm h'ch 10U.A.d otter 1~
1
when t e firs., swimm ng '3togea ( ~ etuluc) ~ ppe r, 'b.c I p r l
conteiut.nr. the more I.Orm l top B1 iittun1n.g .l.a t\.lae~ 1o pm:rcd t) f
e
ot tall bt. tt•~... y jar'3 !'>1 ch are aubs \i'.lt tly f. llcd to tl.
op 1.1
e ~er.. Care ~nust b: taken to . · ,ld carr ·ing over d, ad
bxyos t un...
, e.y~e.. Tall jars a:re up•Hior t\1 (;baJ.lo di3ll•e, s11 o
n 1D c nsi e1·"
9b:!.y redu ed, It is esaentiel that ,.._la.tivcJ.y l'e J.ar
e
e j r
Eurly b1p1nn~rla ma.r tJo obtt i · d · i thout e;.Jc .1 l f cti~1p, but the c L\l '-tree o
Aetnr1aa larv-do must br: fed diatome {prer;ered by JU.flt 0 e ::1 1.tloda, to Jbtoin
brach1olaria or later nt3geo,
References:
----··'Chambers R. 1930 T1e mennel" of ancrm entry in the otf rf s11 rr.,:-. B1o1. .& :1 '8.
Goatellc, D P • .:'9;;o
Fertilization mcmbrene of centlif'l';C At r1 3 Esgo
Phy ic •
Z.:,ol. 9 65 /2.
Delau,er. Y:' j,904. E.le age de lervo p r tno~~n6tique d' a r
a glac· l i • Arch
de Zool • ..;.xper, 4~ <.3c..t· ,;.
1 l, H. l879e
Racht-:rches s~.tr . a fecondation et lo commu cement
' emoRon c. Mem
Sec Phy ~t Hi st. ,'lat. Gene 1e. 26~
Vzy, H J., 1937~ Artie e (p. !H7) j . C Lture Methcda :Lor !nvcrt""br te Ani nlo 1
E.!h ted ·,· Gal teoff Lu. tz et..:. Com toe • j>t,.bl. Co.
;err.m1L. J. J!'~ 19)• .:,
The de
point 1
st~oture o!
the starfl.h, AsterlaJ rube
J
E. 1939
n thod.
<-o uoo)
n
s
~["
D1pl
l.
F rtil':_~~
"''"'"
Bl st
flo
•C
e
l b
for
r
h
t
cloa ng the-blo. c-r•o
tl in th c vi tel i.l 1e
attech d tc th,.. n.. xa
tM l~t<> bl..stula rt
i
•
"
4, .!_;ly f5~~~
The eget . p·Jl::! ore• tllickcna E j
ber,ine. The bl~s pore iu dee 1n .. d to beconc: toe er.t: ,
the animal-7eg~tal e7.1a.
lfl t
Th
I!. '
·'
rv
5. ~!.ddJ.e -~nd l:~.t~ ~!_~lll.!..
'Iile gar. rul1 becomes pe
nl a .d. The b ind ln r
end of the archer ron beco• cs th n"vmlle~, n.. e:>pu 1co
l'lcm thit. end zn .. eenchyme
cella ~Kander out i t the blaeto~o !le. ln a r·Ugh tly
t':Tio ou tpockct · ~
of tbe Uatal · ud bee ne di1t1ne ;r th".: pri ..:>ruAa of t.e o~lo
t the
am•
time, the t•rchen·~.eron bend~ ton,.·~ on:: .side which l:! the futu
vent tl ai·l
Thi.o
ie the first sign of the ch•.luge o radinl into bilateJ·aJ. ayrerretl·y
.N-> .e the ciliu L
in the arctenteron.
6o Transltion tc 11pl3urula~larva,. ' e lind end o1' thJ
towards the ve!l'Z:.:aJborty 1?6~: r m!ll;.. a contact 1 ~11 a
etoruodaou.m" e.nd "..he: mouth bl•t:aks tlu.·o gho '!be tv1o coe ::1 ic
strioted0.ft :from the archei tero·. 'l'he 1t~ft one is'":~;~-;­
!t fo:me a tubule outGr.o th tot~- ol sul 1ody wa~l 1tic
Thin j,e the J?O:re-cena.!_.
y ate.geo
the
Ol
on~
taidt
7. ~.Y tor~,ed r 1pleurula (ep_£~ _Q! ~.£.1~1~~
'ibia. rv :repre enta a
eer... y la.rvaJ. typ oomraon to Ar.tero.idie Eclllno1dca, Oz hi l'oic e
Holothuref
eee Korsche. t. val. l p. 4~HJ}
Stu
ca ~full a v' IJ r l, d r al end l tcral
pre"'erably left) vie
c"""'
••
n
nea'h
Loco
first coot·nuoud a bovc the upp
eet ln the midlin ,
in the uprer ventral art
:trontal field is ch act ri.."tlo o
of the entire longltudinol bend.
illumination, if v llable.
Coelom.
v1e1 ble at the
subdivision of
connecting the
St
vosic1
rly
e
l
first.
Vital sta ..·.11ng wit.h neutral !:'ed ia llelp u'; tut study unatain d o
Study also stained wh lc mount of tlle e eta ~eo.
1m ne
The follol'l ,ng stflges are rorely fomu:1 in dredg a of p; nkt'>n ond d1ff1 ~u t
to rei se from egg•J in the lf'boretol"J. Prepared and et ined
ol mounta will be
provid -d.
8 , _!ul~y formed_B_!p1nn~rie. Conault figureo in G€m ill~ l''.lcBl·i eP nd Agassiz.
This lnrva is cha.. actcrlzcu by a numb .. .~.· of pail·e or lo s or an a .~ ch gro out
!rom the rue.rgin of the ectoderm and which carry the ci.l:lery bn::>.d along. '!'hey m·e
not supported by skeleton. paira of :<me follow eac-'1 other 1:n succession. Youn
eta~es may not ha'c all arms developed~
Arms. Ide·i.ltify unpaired ~n doraal, pairt::d anterior doro~,, Eoeterlor
dorsa!, 2,oster1or l_ateral, ,EE_~torel and Er )~ armn. Seo G«-•• ·r.. 11 y pla tQ lB,
tigo 7~ and MacBriae r p. 463.
Intestinal t~act.
Coelom. The coelomic vcaiclee have grown c \t i 1-t.o long tubea and have
fused 1n ~he anterio~~ part of the larva., .ro further eubdj 'Vl ionE have yet occun·ed.
9, Brachiolaria and met~uor~ho~1 ~tuge~o Study vcrtra
d later l vieweo Con ult
the cxcellen t f j gurco in Gemmill f . platen 19 a~ d 20.
n1c Bipi~ar e or.m~~rc lon~, holJow tubes.
'!he three Brach1olar1.a arill ( trachia) re hc•.::t. They contain diverticula
of the coelom.. They are not cihated but their end d1ok:e diffcrentiatt: fll!lBll
pap1ll~ and can adhere to the substrate.
A sucker, the gland cellH of hich ee
crete a sticky substance, is for.ned bet.,een the brachia. .Brechi! •1nd oucker GE'rve
tor at tachroen t of the larva to the eu bat:r.a ~e in .La tel· L''t age of Jnetamo1-pho ei a
(Gemm1llr p. 250).
1
ne
in d
D1 k
or
r clev l pin
In la t stage of
the diak shrin ~s
eucker and branch!
or details of met
nt
0
The
s
e
p
peaai bl
1 l 0 Ui h
e
A cut ia made e Jl one~quarte
thr ugh both oral
d abo
~ reful y 1nsetc
bore.l portion • ~e or
not to destroy the gonads
por~ion is then pl c d (c
watcb .. glflae. If the en.iri l 1
Allow the ope cd ma e to remain
seminated. The O"'i ries o' the
eggs are uauall. rri. ed 1 th en
the femsle carefully pipe te th~ CF
, r.; ellowinp; ~ c e~ge to settl
and reple
lli1th f eah e
(if the egga do n t readily tr-am f om tl:
ov d with a for cps to a fln~er bo 1 of
strained throup.h cleanr
1
Fertilj 7.et1on
The egg.. o l chin 1·achn u~ ar larger than tho"
(. 35 1 c.·o s
an ompared i th ?b) and Durrounded by a ruuch thlck
.1 lly
in hich
beautiful red pip,mcnt gra ul n re s •apen ed. The e~g 1 t
f the
0 ft' e
jelly. ia pale yellow. E~aminc the nfe1tilizod eg a und r lo and hi~h
map;nification. '!hen inaeninste the e~g.... s ae do c L the c s of Arbacia,
an.d e .. amine the egg immediately aft }r adding tne dilutecl pe:rru uopons.lon.,
Eec£uae of the r{•la .l.vely large size of the ep,g~ tlle fe:r.":illzution 1·eo ion
rnr~y be readily followed.
~embrnne cle ret1on l-roceedu fr n the entranc(• po nt
or the sperm arolmj the egg cor ex in a waveo The m .!tbrenc begins to elevate
in from seven to t •.·enty, two seconds aftel' apeim pon ~trat!.on u.nd is completed
in from nlne to thirty f.econds after it b~;ginso Sil\Ce ep •!n pen<> .rut· 01 occt s
frau fourteen to forty-five Bt,condo after inaf'minat n bot.1 prot;csflea (1. e.~
sperm penetration. aad membrane elevationi may be conplctc within about 40
seconds after insemination (JustP l919)n
Cleavage ot the egg of Ech1narschn1ue i a not mart=edl r iffere t from
that of Arbacia~ and unless thi, fom is or special 1nterent to the student~
detailed drawin~s of the cleavo~e neE.:d not be made. Oue c tl ture should be
prepared and kept to provide plutei 1or comparison wl th those of Arbaciao
Referennea:
Justr Eo E. 1919 '~c fertilization reaction in Echinal'achnitH:I p l'rtl8.
BuL.~
~. 1-10.
Just» E. E. 1923
VI. B1olQ £ull. 44, 1-9.
I. BJ.ol.
I.
H., 1895.
y~
1.
l.
11.
1.,
0. 0 l l •
22.
o t 18 18~
Un1v • .~.okyo,
0
ave~
Job
C
Jour.
l~e 1~00.
Hop~iae
Uni
ogy ot Opb.1o
c i a
·~
H · ey Etllel .Browr. e
t 59.
C ll
C\.1 !!
' t
---~---------0 l 41.. The cy o, o~ of f ... t"til
Arbaoia unntulatao
.rtox Ne e~
l~ l9r
o 7.
H
retnd1ue~
Sven" l93'i}o Ut: l'
; poli, ol. 17.
ckltng
or.
r
•
o.l
ti
ro
• .B\ 1
14-.
rnd cl
c~.
ge of
n euranciac
0
Pu bl. St Zo Zool.
a .. Bridep ·'" ., 1903
F.oy. S c. ..ondo t. e"".
ov J.opme,lt of So:td n . c .. u er
D.~
tu~
Fhiloo. '17 n •
ol. 196 (go,:,d :iJ.. ,n.}
Mort~uae.-"
'1!>., 1~31.,.19, 00 •
n""i:ributio a to tllo atudy of t.ue de eJ.opmeu... on
la:: al forma of Eoh1no el:cr.a., ;r ... rfo !ler1., de .1. ' ..A.cad. ce Sciv Cop nltogen.
~ct. de &::l •• Ser.. 91) vel. ' anr 7~
Mt'U.\.e4.' 0 Joh.
AbO.
Y6ni~lo
l84B.l85;;.,
Aknd. W1oa.
Uebe:· die Lo.rv nunc dieM teroorlilH'c der }~ohinodcrmc
Der~in.
Sch'llidt~ H~ ~ 1904.
culatua..
Zul· Kennt1.'~V dcr Larvene1 twicklung von Eohinua miGrotu er.•
Vel':'h., Phya • .,med. Geeellecb., Weh z'bt rg, voJ.. 36.
1'0D Ubiec~he J~.
1'33'7., Die no:rmsle SkelettbUdulg bei Eohlnooyom a ptsillua
und PfJ&"lll1ecn~.nus mUie:riso Zei tschr. w1as. 2:C.ol •• vo.l~ 149 (gcod f.l.gs. of
origin of la~·vel skeleton).
oodland 9 ll., lQOD,. Stud.Lc 1 aptoule for net ion. Iii.. On the mode of
fonnation ot' :he ap.tcula:t- ekal ton in the p u teu or Eohinua euculentue.
Qnert. Jour. Micr. Sci., vol., 49.
c
p
cr1t1 l
0 1)
Needham, J. 1942. B oohemistry
re 1ew of th; iochemical a ect
Schleip,
{Complet
!II ..
w.,
bi
-
Determin t.
eo veri,
0 l
r Pr1.:ni 1v
Detennina tion
iop;raphy of earl1el' "ork up t
l 29.
liv1dus.
Hoeretadiua,
and b1lat rali t:t_.
or polarl t
'l'h .. , HlOl.
ce~trotus
s.
Die Po arit t von Oocyte, Ei Ull
Ana • Ol.o l •
~ol. J hr • A t:
d
s.,
lna tion d r
und Wolakt, 1936. Studien u~bc1 die
Bilateral S)'rronetrie des jungen SeeigeJ..• .Keime • Houx•
Lindahl . P. E.P und Stordal 9 A., 1937.
der e~rsten ll'urch<.' 1m Sceig .l.e1. Roux'
rarx . il.h.· 1931.
Arch., vol. 1.:.5.
Zu~
Ueber die
rch.
. t ongylo
• t
v .l..
l3~ •.
Det rmin tion der Ri
Problem der Bilater litat i
Scelgclkeim.
btun~
Houx'
Pt.or~en; T. Ho
and l.yon, E. ?. 9 190'7. The relation of' the oubntances of. the
egg ecparateri by strong centrifugal force, to the loca.: on r the embryo.
Roux' Ar~h., vol. 24.
~rewmao,
H. H", l'.l23. An eJtperimental AnHly ltJ of asymmetry in tle stlirfJ.ah
Patiria miniate. Biol. Bull., vol. 29o
D. c. p 1 941. Echinoderm bilateral det liTlinC!tiol'l i1 c.t • 11ca.l. c nc ~::1~ra tion g.l:'adient . 1. Jout'. Exp. ZooJ.. f vo . 86.
Peaoe~
-------~ 1942.
IbJ.d.
n,
Ill.
Op. c:it., vol. 0\l.
Tennant, Taylor and Whitcker~ 1929. Ani.uvest~gation ou or3aniz.ation in oea
urchin eggn. Carnegie Ins. Publ. Wa~h , No. 391.
Wilson, E. B., and Mathews, A. P., 18\.15, MntHation, lertiJ.t:z.etion and
polarity in the .Echinoderm egg. Jour. Morpl,., vol •..lO.
//
1
r
w
nu
r
1
Q B
14.
en •• v
Q
o."'ra
ol
:n n.t
t
b
0
... n
l'
1
L n
U.C'
gi
ge
ucr I
ra
l.:l.J
Lind hl Po E. und 0 an,
l ge de= Vege~ativi i~rung
rcho
ol. 140.
Rulon, 0 , 1941.
and Cn~free ater.
• o •• 1940. N ue itrag zur phy iologi chen Grund ...
,s Seeigel-K 1me d rch
thi
To en. P.oux•
ion or d
opmont in the sand dollar by
rlyeiol. Zool., vol. 14 1 1.
•odi~ica
CNS
Runnetr
J
1914 and let~r~ An lytieche Studien
ar di s 1ge1 ut ick1un
Many papers chiefly in Roux' rch., vol. 40 41, ~ 46, l05r 113, nd r • r.
Zool.
on Ubisch, L., 1925.
II~ III~
Ze1tschro
Ent icklungephy iologische Studien an Se i
iss. Zoolo~ ol. l24v
1931-1937.
Jntereuchungen
ber Fonubildun mit Uilte experiment
~rzengte= Keimblattchim!ren von Echin dermenlarven.
volv. 124 9 126 9 127P
on transplentat on of
Hoerstadiuep 1939.}
129~
1' 1,
microm~ree
-VIII.
Roux' Archo
34, 137. (A critical revie ~ thla ork
be .ween different species m y be fo~nd in
- 1 -
It ··as discov red·).,· P =' rt (18.2) t11ot. ·. .1cn ~L "'r1,
stat:;es J..:1 f!:>' - Tl c'1:.r.. d€.\·el_ . , n!· e~·e n1 rr~it. od t~' PoJ,·t5. r:.~
of :;_i·;:;·-- ;m c 1~ f;'.lt of'·c""l !'.. l~. d
) ir:v..
t··
::::ctcc'c ...·:.:t tc· ,': on the c' •,::.:..• ,c-~··"!.'5.~ 4:icr f er., C'('~(':'l ~ S 11.~0 ·i;' • s
ti:·.:e 1\L'Cl·Cli.S G['ents : "- \'6 ·-~ _ 'C."'C( tc .._l' C. 1~._; -1-;'!i:-' ( f'f ct ir:
ec·.5:1oc1c:::·:s Pnd no~&'e ot' t:::.• fo:.:;:;s •
.A J.J. s t
)J'E.
3
o~·:.-::~r,ined
one cell
·1o.t r .
1· lace 2-3 drops of ccnc 1 ·ntl'2ted O"".,fl L t_. l ·'-'
Ex, ·;1j.ne c.t t ' .e e~d of l·J - 12 :.:01J. . 'E1 r.nd conpar~. •i ·.'l Cl yl·:.·ols ln 1
mal sec-. \;e.ter fr·or::. t:r~ 01 '-":'· ~l-1 ')~ tc::> Lf c . . . , .., . I'' c
'-'- orc·Ol
of ·-~"e e :f.s fr01,., t:1e 1-i·c· 5.u':'! ~- ol.1·cjon to ren w[l·;; c.• ''· .1
::.oe t
t:"n Cl' 2S li::tle cs . .. O~fi- 1 }C of ·c~e ]i·V•i;;u.l co}i.~·'·ion .
1'•
J:.'C C'nd of 21 ~",)U~~s c..ga~ ·~ C'Tt:minc t11c l l' V F .::: -' o
s·)ln.1.~_cm <r:c ·;~:.."'rn:::i'c-··· a por·t ion l.o rcr "h.:t-c-· .
Co·::pere
trols . ~·- r.:ninc !l!.s c t· o.:- c l c rv[ c 'h=' t l'r ve 1~ocn tr"n. "c1~" ot' to
S€..?. '.'ntcr 12 ~.ouJ~r ··r ': ·onn :-,r .
A:- 10-12 . 0'.ll' ::.r.:~··t.l'V<.; [.
r
01'
t~
() nv:d. 72
'1()L't''fl
OJ~['.,
".nt ~-nc.
maLe drn•':Jnes of L!t: J.:..t>Ju~.1:--,v~·.c c1c1 control~ . No;;c c,j,·fu~J..y
i.:..::: ... f.fc.cts of inj.tir:!.. ·''1' c;. Tl.~i:mcus C.XlJ O"' l'G to Jit.lt1.:..,.rr. .
Skc.tcl~
aJ.l c,sbPcos of entor:l_el·,i.zcc:(_' .., .:o.16 l't..;COv~l,y . 'vV'!.c'C
.. -'>·l";.E to
t~l(; e. __t cc.l rc sion of c' ~ 1 c.rvc?
Ro:fGrcncGs .
Cr:Ud , C . M. 1936 .
A co'1·i ·· 1.'"'..1.t5 0'1 to
t~~ ~~~siolo(y of
J-.rch . T'nt11 1 ..,,c c·.._ . , J.3b .
He:. bs t , C . 1892 . !:.xr-'' _•j:·.-:.c Tcc.ll; Unt"r:,u c' >un,-::;n n u'hc r &t: n J~5.nfJu3 s
dcr v orLndcrtcn c~t8iLc~ ' n z~~~v~ no tzung dcr w1~CJ 11Ln
J:cdiums c.uf die Entv•lcklur. . g dcr Tiuc .
I.
Zc:l·i~~·c>l~~ 'liss .
Zo ol ., 55 Q
Linc18.!:1 ,
• E . 1936 .
~1..1'' I:t..,...J.i.ltn:i.s civ::• l/':rniolo~i~ c',
Cr:;.,~.mn:t:tf,Cn
Gr~u:dl~gcn c1 .: r Dec :i.'l"n.t·:"Lon ir:1 <'l.<..i [~... l.k<... in .
Ac'.; soo:1 ., 1? ...
J . ··: . J.924 . An t· "Jt,l'ir.· t< i :::-tur'y t'i1C ~ .tlr·~.i.r.lo··~c-.·
·.n·: :·.1.-Y· to t.J..on of r::x o <:.c:<-;:n l:... l; L0'1 ~ n( ··""]: i.~cd :' ol.r ' ·.C'' ·ci c· ,r
. :l. ol . B l~.: . , <G .
i,:P_GA.r·t l,J.l' ,
-2C:C:NTR IF:JG.t.TI
:r
A ·n . ·=nOGOJY I:
ARBAC IA
By enormously increasing the gravitational forces it is
possible to separate more or less completely the contents of the
e0g of Arbacia into layers of substances according to t~eir srecif ic
g~a 1r~ ties.
The layers in the sea urchin egg , after strong c cntrt·
f·:. r..;.::. ·::; i on , are fro.n light to heavy: (1) oil, (2) homogeneous 1' ,t t~.·~r
:1.9.-r er ·;Jith high protein content, (3) mitochondria, (4) yol ~, ,,igm :-·~17. .
The nucleus is throvm into the lighter regions 1 ar , ·
~en t·_'ipetal end .
Continued centrifugation •ill cause t he egg ~­
t !'e2.k into hyaline and pigmented portions . The pigment d por ... i. o ,
a l 'though without a nucleus can be fertilized and will dcvelo}- \..: t.c:
r ---· ly ti.1e sperm nucleus.
This proc dure is known as mer0uon·r . J l~ ,
,· :::-,1.1 r_: lea ted fragments can be S timula t c.. d to Cl('D. V C thro 1g'.. tht 1.....~ ;)tl
:''C ,arthenogenetic agents.
Examine the centrifuge and understand its mochanisn~ . Fil}
t .:e small c entrifuge tube with 0. 95 1! . sucrose. Now with a fin > ;r
di'D.\'n pipette add a quantity of unfertilized Arbacia e ggs to tho
sucrose solution and insert firmly in the arm~ of tho centrifu e .
Ce ntrifuge at high speed for 10 minutes. Renove the sur;ar anu
suspended eggs to a syracus~ watch dish of sea water. ~ ash with
s e a water by drawin g off the sugar s olution and adding sea wat··:_ •.
Note the appearance of the centrifuged eggs and egg fragments l~ ""':.''c :r
t!:·,_t., mic ros cope. Draw eggs of various shapes and vurious fragmen t c: •
Eggs do not always separate exactly at the center so the fra~1o nts
will contain various proportions of the different substances.
Add a few drops of dilute s perm suspension to tho stratified
eggs and fragments. Note the time and regions of membrc.ne
ele vation. Watch the cleavage pattern . Study and sketch the
de velop ing fragments in va ri ons sto.ges.
If time permits artificial parthenogenesis may be at t empted
on tile stratified eggs and ogs fragments.
Rc.f e r once s
Ho.rvcy , E . B., 1932. Th e dev ol.Jpment of half and quarter eggs
of Arbacia punctula t a and of strongly centrifuged whol e
eggs . Biol. Bull., 62.
Deve lopment of the parts of sea ur chin cggc·
separated by centrifugal force . Biol. Bull., 64 •
_ _ _ ., 1933.
. , 1936 . Parthenogenetic w~ rogony or cleavage without
----::=mcle i in Arbacia punctu:ta.to. . Biol . Bull., 71.
Pease , D. C. 1939. An analysis t>f tho f a ctors of bilate ral
dc termination in ccntrifug,; d c .: 11.inodr 1m eggs . J. r.xp .
Zool ., 80.
Us G abovo for other r c f c rC11 ';vS on centrifugation rmd
merogony.
N ot e ~
- 3 S :s I 1
J.:
mhe fo.ct t';.a t mf, .y eggs can be ... l.i... lla t~d t
e , o . i thou t
the sperm ~2s at~racted t.c attentior. of msnr invest ators . One
of the mc.:st iv:I;'::)r"ta!J.t of tL-: e£rlie!~ wo '"Cl'S wa,, -r- '011 s :i:Jocb ··hose
forr:iulae for the treatJ.e-:-l"i.: of eg5s to ir.it::....tc d v~..lo". 0.1t cr in
c·trren t
se •
t
In ordGr to dilute t'be rctivr.ti::tr co .. ttion . . af' itt~_( as
r..Jossiblc : 1 d:--y~ 1 ur-~f~,r~ilizcd eggs sl-:o·~~d o usnd . T c.y r.o..·.v ho
ol)tc.. ::.nc d i~ -::.:w f :> .... l·n:ia~ :..~.-:-.rc.cr: ··,r • s:1 tbc; s c,"'. urcl.in t:it,a'J t:l'} y
1.1 .dc:c
V10 ;~rosa ·Hai:.er ~.:,;:::y.
_.~.so sue t.h ... t y ·1.r h:. nd.., cz d :h:stru
BEn-es rro ·voJ.J. rinsed ::n .trct' "!2t01' 4 · ,.ii.;h VQUI' ~lCLC.OlS cut Ci.J.:;.':O·
fuJly ~rou~d the test just ~ tsidc -c~c p,rist ~o uGin · c ru not to
L!;:ure the ovrrics . :\c.1.:c~vu • ::.1· i~- ~~omo t.nd J.c.nt rn . nd
co< Jamie fluid . Pl~.co ~h.: ·.nili!L.J. on its S.t,i h1~' o:-: '·' ~
f:. e& i:1 :. clonn finger bO\ l .
If ripe, egg,. •·;il~
,
t'11J [,onopor~.:s in c. conccnt:cc ted mt.s8 ·_,nd frcv of p :..'1 v :;,cor .J.
f'J.uid .
S 5 nflC tl"(.IJ. b~Gnt o
~!.0.00 t < drop Of dry
g(; in t;~ C 'Jot tom Of
v1ccc1 dish c-.r..d flood 'id.·c:: 2 . 5 M. N~ . c1 . /~~.tc:~
Ol a clock ~ nd [. t 10 •J \.. CCi'lC: :i.n·l;crv.. l~l 1' Tt10V .1 ~· !: ort:i.O
ecr~s to 250 ceo
of norm2.l ~ea v;r.tll' • SL~ di f"';::• ~,. yl_, cx.voruro
pc r i ocls sho·,J.ld be [.de. q_uL ·(~~ • C'!cck ca rE:fuJ, y tl;e expos arc e riod
b•st suited 1or mcmbrD.nc elevation \'i.t'--..out c'rto1 "~,~~ :l
V.. ry the
cxrosure time bv 2 Ec cones intcrv~ls on either sid~ of this e1iod
fOJ .. 100 ~:; munbrr.nc t l cv.l.ltion a nd cJeavagc .
<...
"
<
Double t rca tm.ont . .t?l8.CC a dro~ of drY I,. r;· ·r· in ·l_.'·.c 1 ott01n of
n. v1nlcll dioh and fJ~od V'J.t::. f'. sol.:ttion of 2 , 8 cc of 0 . ~ N hut;:.,.,·:c
acid plus 50 cc . of ::ocu 'P;::...·t-. t,:: . R~:.r.1ov~.... c.g']:r- fro1~: t~1i~ :-.oJ.utlou ·c.;
~ca 11ater at 20 , 30, 40, cn.:1 50 minute int,,:... . v c·lf.. .
Afc.cr F:: min ··
utos in ~en water remove the cgsG to ~ ~olutio~ of 8 . 0 cc of 2.5~
NGCl pl us 50 cc . of G\..8. ,_.~t::::1' . A:it-.r 20 min:tten :i..n Gll5.~J ::.vl,JGr'Lonic
solution roturn to r•ca wr.tcc~ . \·,r:.tcl~ for m,:mbl'\... nu ,·J..._v( tion ~.d
c 1 .._,~- vagu . How fo.r will L' r vc c obtr invd b;r Yi[:rt:i:' .nO['l ·1.u 1.~ic tr0c t.m~.- n t d-..v.:..l op ?
RC'fcrc n c or; .
Hu:i.lbrunn, L. v. 1915 . StucUcn in crtificic.J. l)art).cno·· n .. L;i~:;
H~ys lCD.l cho.nr;cs i:1 the ugg o:t Arb1 c L ..
Biol . BuJ.J.-, 29.,
.
II-:
Just , E ~ E . 1922 .
Initiation of dc v lo_pmcnt in tl""~.c. q~g of !Pbccia.
I . I'ffoct of hype rtonic sco ··'No.tc":' in r,roducinr m mb:> no______
sopor~ tion, clcnvncc &nd to - swirumin5 plutci . Diol . Bull ., 43.
--..,..• 1 939 ., Bo.::;ic Methods f or Expcrim( nt~ of Eg:;~ on rr.crinc
Anima l s . P . Bl[ ki::. ton' c S ,:mu & C.: o . Philt.d clpllin .
Loc b , J. 1 913. Art ific i~ 1 J. ::.rt'lcnJf';"ncn i:; nn<l Ft rt iliz,_t Lon .
Unj. v . Chic L.[_I'O FroDs . Cl,:tcago , Tyler: A. 1941.
Artificic.l h.rtlL'1.0{!,l-nC3in .
Biol . R(v , V. lG,
- 4 4.
F!.:RTILIZATIO l :
2.
Conce~tration
(Arbac ia recor..mer.ded . )
of spermatozoa .
Add 1 drop of dry sperm t o 33 cc of sen - 'later . T'ni~ is a C. l;o
(ca . ) dilution of sperm . Prepare eggs and place 2 dro~ s in 5 d:s1:u!3
c ontair..ing 25 cc., 50 cc . , 500 cc ., and 1000 c . of seq-: ··or r·s ..!··
ti ve ly . To each of these dis. te s a ad one drop of t .. l ~ • :.r' q.>m', .sus··
pe~s:..cn and c .:;.lculate aft' r f::.fveen mi::.-...1·~13s the m1::a' ""r C crr..:c1)bn 'e)
of :;:1.e ee;;s :>·rtil::..zed .
,_·SC!"i" .:caL.:. "ft.er 5 ho . .trs .
J.t t.11e " i.:~:
t1me Lal·v a l'' SIOl·:n St:~r---·-sil'~:- "!'.d c,dd cne d~·o:J to Cf:r'"!-3 i. :0 .~·.,
25 ccn, SO cc,, 100 c~ . _, c.nc!.: ·J cc. s ~;n ..·\•a·c'"'r. ln t'".'J 1 . .\"t uer.1 ~
record not onlv the J:-er.:e~-,·c c.(l'r -J.f t._e e.:- "s fc.~tiliz ·t' but l:o thr
de ree of 1,oly~:f.Jt..rmy in S<ICh J.nstcncc . . . (:Jot0 fu•st clc e1v c .. )
b . Effect of hydrogen ion
conce~trl:\tion .
S e t up a series of f1nger bowls cont"inin ~ 100 cc . or Lolut icn
as follows: sea-water plus 0 . 1 cc ., o . ~ cc . , 0 . 4 cc ., end o.~ ;c .
n/10 ra~H , and 0.1 cc., 0.2 cc., 0.4 cc., v.6 cc., 0 . 8 cc ., and
1 . 0 cc. n/10 HCl. This makes ten dishes, Add anot:1er dish contalning sea -water from t he taf.!. Add 2 cc. of eggs to ea c h dish
end examine to determine the effe ct of each solution on the eggs.
~epeat to determine the e ff ec t of t he solution on the spGrm .
Ro poa t once more to determine tne effect of each solution on fortiJization and cleavage. In testing th€ eff0ct of each of the solut ionc
upon tho gametes of one of the sexes alone , obs Drvo the gametes at
in t ~ rv&ls up to one hour.
The ~ydrogen ion concentration of each
of the s olutions should be determined by t l:.e indica tor met11.od. Who·~
conc e ntration of sperm did y ou use in making the ins eminations? It
should be the same in eve ry case.
c. Cross- fertilization.
Prepare eggs and sperm of the starf is h and sea-urchin. Im;e···, j nate the eggs of each with the sperm of the otl'1e r. Observe the
fertil ization and development of the eggs. Analyze your results.
What effect does the pH. of the sea-water have upon the readine~l-3
with which the gametes unite!' What effect has the concentrat1on
of the sperm? What percentage of the eggs develop as comparee. 'v5. t!'1
eggs fertilized by sperm of the same species? Is there any v;~de.~10c;
cr patroclinic influence during the pericd over whi c.\2 your obu.. rvations are made? Significance.
d. Effect of agents in solution on fertili!:ation,
To sea-water in measured amounts aad definite quanti ties cf
coelomic fluid. What effect do these solut ion s have on the Gggs?
On the spe rm? ,Inseminate eggs in these solutions as dcscr .... bJd abo./<',
Nov: insc::minate eggs in sea-wator and, aftGr i, 1, 2!, 5 , :1.0 ur:.d 15
minutes , add to the solutions of coelomie fluid. Docs tb r. prGuvnco
of coelomic fluid have any effect on the fertilized C/3f5D in -'.:;hf..i co~
cr·.ntra tions employed? Analyze.
Repea t this experiment using in plac e of the coelcrr'J:l o f:p·,ict
:)"!.u.s sAa - wate r, solutions of CuCl2 from n/8 , 000 to n/80 , 00'J mo"'..: ~n
.:;;, . .. ,(.;.cn r .
I s there any similarity in the resuJ.c:::?
- 5 -
5 . CI.EJ._ .. J;.'}E:
------
a.
Effe~t
of
te~perature
on development .
Fertilize e~gs of A~bacia and place in seu~vater i te· ~ers
s··-lOt-:.ld be k~p t at an even t emperature . T-:e tem erature ::l1':·.'.•-:.
::10-~ ~Tar-; a de ree dur i ng the e x pe r i:-rio:!:1t.
:.tt8.;:!)c to cbt"l.i.l. 7'.:'C') < :1
of t!"f, ~ime a7; vJ:~ich c lea\.·s.ses (l8t, 2 ,f. ~~... r:, rn r- ch) t1i·e. ]:18.( 8
in sevr~rs.~ 'b'-' :~ hes of egJs . J,,a).:e re c c~".·:s oi ;4s ,'"l.ny tem n''a1'.''"";> ...
possible ana •'Omp'.lte t~1e temyeratt:..re c~.-.t.f f i c :.ent f or tho p1... :::><. .....,_:,
·.'l~j,i~l".
b u Ef:·e c t cf osmotic
~1res s ure
on
deve.~opmer..t .
Bo i l 100 cc. of sea -~ater co 50 cc . The sal ts in the senTater will be twi ce a s c oncentrated (ca .) in such a aoluticn a~ 5:.
sea --water . N ow make up diffe r ent s oluti ons us ing this r 1 edium tcropres e n t 2/1 sea - wa t er , 7/4 sea -water, 6/4 sea - water , and 5/4 S€'~t .
water . I n the same series make c once:1tr at i ons of 7/8 sea - wate:::-, '/H
s e a-wat~r , 5/ 8 s e a-wat er and 4/8 sea -wa t e r by diluting the l:!eH-··mc;,~:
as i t comes f rom the tap with d ist illed wat er. Int o each of thMJ·~
solutions place f e rtiliz ed eggs of Arbacia (5 minu tes after ins8tnin.:t·
tion) . Re cord the behavior of the eggs during the first three uiv isi o ~s .
Time of cle~vage? Percentage of cl e avage? Extent of
deve lopme nt obtaine d! Type of cytolysis?
c. Modification of cleavage pattern.
The cleavage pattern of the egg of Arbacia may b e alt er ed by
pr e ssure. Place some of the fe rtilized eggs on a slide and wi t h ··
draw the water until the cover pr e sses them slightly. S e al the e::ie-; ..
of the cover with vaseline to prevent evaporation. The e ggs s h o u::..c~
b e placed on the slide approximately 20 minutes before t h e fir st
c l eavage. Observe the cleavag es of the egg through the 8-cell s·(., ~~,...
Return to sea-water in a watch glass and allow to develop. Wha·;:,
effe ct does such treatment have on the formation of the embr yo<? Fcp e a t allowing the egg to remain under pressure for l onger per:i.od:J
of time•
6~
DEVELOPMENT OF ISOLATED BLASTOMERES (ARBACIA).
Fertilize eggs of Arbacia. About 15 minutes aft e r fer: LL "'.. t.<. •
tion remove the jelly and membranes by drawing them into a vG ·.'"). fJ..r:.(:
c apil lary pipette with a bore 2/3 the diameter of the egg mcrro.r'F!l'..Li;'.
Tbl s may also be done after the eggs have cleaved to form t he :u:; i;
t wo b las tome res. With a fine glass needle separate the t wo b' 1•• D t 'J · ·
mere s while still spheres and remove to separate dishes. K00p
a cc ura te records of the behavior of both halves of eac h err,g during
its ~ubsequent development. Analyze:-----7 .. DEVELOPMENT OF FUSED EGGS •
Fe rtilized ova of Arb acia may be made to fus~ by the fc11ow~~r
method . The results obtained enable the exp eriment er to c'l.rA:.'. certain c onclusions concerning the organization of tho zygoLe .
- 6 -
Place fertilized eggs of Arbacia in the follo\ing soluti ons:
45 cc. sea H2 0
30 cc.
20 cc.
11
"
a
11
55 cc . m/2 NaC1 2
II
,....0 cc . II II
II
80 cc. II tl
lr:. -si:.e.~-:> solutjrms eggs fuse or a ggl·:.1 tinate.
Ld~.ve in the no!.ttt=:..:Jt,
& i,.) ::; ·_ours u-:-1d then t!'a "· sfer t o p .... .r. sea- ·~'. ter a"'ld o ;-;r,1 ··~.: :··:..i..·
SS-l'.F'.• c dev'3 lo >men~,: .
The .:usions chou.:. l be c:: ate:t a 1.r.. tL vidt i:.:
a:::.a r .-.r; col ~ .:-.:.ively from early sta ge ~ . Anal :n~e sour re . .~ ·~.ts.
RemovE. the ripe e!~·ss f .~om ~~ .:.eE. urchin or SP.nd d :i.la... cc
sea rvate r usiEg all pos s1ble care t::- pre v ent i:tsemi nation . 1 r,:;>.
wit~ s'3vora l change s of sea wa ter and test a few f or f crti l1~ -.
N0w 1.1.s ing a medicine droppe r and with a. minimum quantity of sea
'<iB.~c:· trans fe r approximately 200 of the unf e rt ilized egg"' to tl:e
following solutions in cle An fing e r bowls:
(a)
(b)
(c)
(d)
Normal sea water .
Calc iwn-free sea v1s. -te r •
20 cc. of 0.54 M. Ne..CNS plus 100 cc. sea wate-r,
20 cc. of 0.54 M. NaCNS plus 100 cc. Ca-free sea
wa·G,.,.r,
After 12 hours in these s olu ti ::ms run the eggs through s e~p ·::-~ .·
f er tilize in sea. water with a drop of
Examine for fertilization membranes ,
Co•;·e r the finger bowls and, if p os sible, leave on a wate r t able<
c}~anges of se a wate r and
d~l~te sperm suspension .
Examine the developing larvae under the microscope in dep~·css :.on alides at 12 - h our intl'3rvals following fertiliz at ion.
C('J,, ..
}~e.re the controls and tests.
W.~1at is the effect of the thiocy a:-.~· ~ ,,
C"1 ~·iabi lity, cleavage, gastrulatio~ , etc.?
Explain . Wha t pr:-
port.Lon of the larvae show the effects of the a gent?
sentabives of t he cultures at each observation .
9.
EN~ODERMIZATION
Sketv~ :·u~'I'J·
-
OF ECHINODERM EGGS .
--.---
Place 200-300 f e rtili ze d sea urchin or sand dollar cg~~ L~
(This s olution is made up b y dil1lting r. ~·J> ..J.~··, ..
n ormal nE:a water.) Arrange c ontr-ols ··-•·;
:·1o·.·m.?.: nea water. Keep in c0Vered finger bowls on a "vJat.cr t a':..~~~
J.:t· 1v.:.:rjsiole .
At the end of 4,8, ard l2 hours examJ.!'ie an6. ror!./.rc·
3. ps:"' ti on of the larvae to normal ::~E:a water for fur cher d3 v e.·e ~·;.~ . ·
2~0 -~c . of M/30 LiCl.
30~.T~i on of LiCl with
1:' P.ll '[. •
Examine the lithium-treate d and control larva e nt ~PPI'C>:' ~
in:flt'.'l.y 12-hour inter·..r3.ls for the next thr ee days. What, .i.'3 tho
e£fec·t of early tr eatment with lithlwn on invaginatlon ·? r '1
mes enchyme formation? Vt/ha-c h..appens to the presumpt:;vt, ryt oclo1.o.?
Ectode r m? Explain . Sketch larvae in varying stacc& of ~nhib~~ion
and re cove ry.
- 7 mljl;... '~y
·- .l •
•
Flnco 200 - 300 fertillzed sea urc.i. or sand dol~2r cgrs in
Ca - .froe sec. ·.;ater (DistilJ.ed v1ater , 10 , 571 cc.· l!aCl , :5~..-7 "'.1 . ;
:~cl, 8 gm . ; rgS0 1., 66 gr: .; l!a:.oo 3 , 2 g1:1 . ) .
'1 n controls in
normal sea v:ater :
Observe at 4 , 8 , 12 and 2~ '.lours after fertilization? tt each observation remove a p ~r tion of the eggs to
nornc1l Ge a wa ter and stud~ at 12 - hour intervr::ls .
Sketc ....
re~rase~ ta t ive forms .
Hate the effects of early treatment with Ca - free sea water
on c}_eevage , formation of mesenchyme , oral lobos, ~.GlaJ nrms ,
sy:y:r.1etry , amount of s ~eleton , etc. Jete the fusion of sane of
t!2e larva•.
VJ.w t a re the effe cts on fields , pr, tte r ns , etc . of
su c>~. fusions. How would you expJ 2 in the r.c tion of C - fr9e c• • ['
'·ater?
11 .
SUSC:fTIBILITY GRADI:::.N'IS E1 ARE.!CIA A1.D ASrfF.RIAS
-- - - · - -- ------
J..c.. ti vi ty grad i ents in t~.J.e developing ore:;E>.nism l1a v0 indicfl ted
a me c:·,_anism of correlation e.nd intt:.rration ,.,J.,ieh. i.s of n nyn:?JnJ.c
nature . J.<.egions of great er .~:-L·:rs io1ogical a.c ti vi ty may c.xcrt a
central ( dominan ce) over regions of lower activity .
In uncleaved
ec;8s the ?.niraal p elle commonly sh.ov1s E' e~reater rate of ac\..).vity
t . Jaa t:1e vegetaJ. pole.
As development proceeds se c ondar:r r< ["lOr r~
of ectj.vity appear probably as a. result of physiolo~~·ic8l iso1rtlon
from a .i.Jrimc.r y grs.dient sys te;11 . Regions of higtt 2 c t :l vi ty nrc nore
S'\..,sce1-ti'ole to t oxic agents than are regions of lov er octivity .
T~ey die fas t er and , on deat~, c ytolyze or distntegrate tJrus mrrki~G t~eir
positi ~n.
lJ.ac e a quantity of un:'~.-:··tilizcd egr;s , blantu3ec , ['t•f'truJ.c.o.,
c:.nd pJ.u'l.:;ei or b ipennar ia in '.'<:1tch disbes . Remov e. ~u· r.mc'·, of t}')O
wete~ as p ossible a nd floo d with sea water solution of 0 . 01 M, ,
J . C05 H ., and O. OOJ. M. 1\CH . rl'J.·.e wo.tch dishes s~1ould '~-~f. covered,
J..·c :i.n·cer vals of 30 minutes ob:H.rve under the low r..~m1er of ~.,~10
r:1icro3 cope noting re g ions of dis integrat ion in the ec;gn or larvtJ e
r:~.nd t).:•.B COmparative SUSCef;tii)ility of J.arve.e in diffel"Ont StO[i88
m· devel op·men t. It is often e.dviseble to follov1 the 1 n"cce~jS of
disintesrcti on thr ough a sin,0;lc egg or lrrvc. • Skctc !~.es 1i1t,..1e
O.'U':i.PL,. tt1e d i fferent }!er:\ocls in t;J.c d isi.nter;rc.tln[ ~·:COC.Or'f~ f.!:'O
advisable . This ex~eriment mpy tvke sevcrnJ. hours .
Gradients in reduction of vital dycG ~nral l el th0 disintogre tion gradients . Eggs or l ar vae stained J i~htly with Janur rr~~n
(1 1-art to 100 , 000 - 200 , 000 ~,c:.a.-'lilate r) for o. few mLJ.uter. ancl
seaJ.0d under a coversl i p wit·,., vesoli'lc ';•ill :..'educu the c'yc ·i;o
a l)r:il1iant r ed v;l1en the oxrc0n iu dinnntnrJ.cd , Tho mo:c,L o.ct.ivc
r,., c_;i0 1J.S , :lf uninjured by t.'·c (1.:JG , "' iJ.l b<.>comc red f lr[J t . lJhi~:
expe:c·1.11cmt requires c onsid.:.:.'u 1./~
ctrc. in ~,t ..-injng rncl U:J uJcio:Jel .
OnJ.'r cl..,.c.r or liphtly pie.JE(~nl;cd c.c:_[: and lc.rve.c m:o ~mitt '.J.c .
-a
Refe rences
ne~~t.
;j ·. <.}_C., C. :·· .
-J.~
1913
Axial c r ad ie:n:cr in the E>ar1y cleve1o
t:e st&rfisl:. . Ame r . Jour . of r-l!ysic::.., 37 .
• 1916 . L. ia1 sus~eptibi1i~y grad i en~D i n t ~
·devc:' opncnt of the 5 e a urc_ L • E5.ol . BL.ll . , 3C,
B3~ly
• 1936 . Diff erential redu c tio~ of vi tnl dye~ in ~~ e
· ·-· -·--ea:-1y development of ec'. ino c1c.,~:, Roux /.rcJ:1 i v • .• .J. :.5<
• 194.l.o Patternu 2nd Frob .... ·.ws of Deve..:.opment .
· -·- ·-- nive rsity of Chicago l·ress . ~ilic cr o .
.1:'-le
1
1944
~:perimental
Enbr rolo~:
of Aru e ll.da & 1oll1.sca
TE..XTBOOKS
e er
m:ley & de
'.1 or[San ,
1
T. H.
eedham , J .
Schleip ,
''I·.e1
• ss,
1934 .
Elements of E..xp . :&!! r ,·oloc;y . ·o~~cT' 1llnn
1927.
E::perimenta 1
U . Press .
Ebbryolo.~~r .
1942 .
B~ oc r.emistry
nnd I1 orp'
1929 .
Die De termi n8. tion der Primi ti vEnt,·Jicklung .
'\.1 nd .
VeJ•l. Leipzig .
1939. Principles of
p_ •
Snecial
Colun1 in
o~~..;nesis . ~ nc~.'iillnn
D(!~_,eloprlent .
H. Holt .
~e ferences
1 9 17.
Effects of centrifugl\l force on the
structure and develop ent of t he
eg~s of Crepidula. J . Ex;:> . Zool. 1 22 .
1938 .
Dj sorientn tion of development in
Cre pidula plana produced by low temperatures . Proc . A~ . Philos . Soc., 79~
Costello, D. P.
1938.
Studies on fr .qgnen ts of centrifu~od
Nereis eg~s. Bi ol. Bull. v. 75 .
---------------
1941.
Several ahstracts on isola tion
experiments of bla stomeros of Nereis .
1\.not . Rec . v. 78. Suppl. , p . 74, 132,
133.
n. E .
1896.
K:.p . Studies on Gastropod development .
Roux Ar chiv. v. 3.
Conlcl1n, E . G.
_______________ ,
Crampton,
Foerst11dius , Sven . 1937.
Exp . on determina..tion in the early
develoument of Cerebrntulus lacteus .
Bi. ol. Bu ll. 73.
Lillie , F . R .
1902.
Differentiation without cleavage in
tre egg of t'le Anne lid C'. . ne topturus .
Roux ' rchiv . 14 .
---------------
1906 .
Observations and experiments concerning the e1enentiiry phenomena of
em1Jryonic de ve lopmen t 1n C haetopterus .
J . Exp . 3ool., 3 .
Mor~an ,
1910 .
Cytological studies of centrifueed
e~~s .
J . of 2xp . Zoo l., 9 .
T.
H.
-2,.
'k 0 ft"
1. OVl.
-
1938 .
Er:bryonic ·c tcrmim. ti o in t :1o
.nne lid, S .be11nri:..~ vul .ris .
1 . T""e differenti tion of cc to derm Y ."'en se 3rn ted t! rou 11 1.nd1ced exog[\stru1'ltio•1 . 2: 't'r:ins l 1~n tn tion of noll:lr lo' cs nnd
b18.sto!1'cres . Bio1 . T'ull . 74 .
--------
1939 .
:,c > l1B tion in .:os, ic e_ps . -..ol1ect1n:> ?:ct . 14 . 8 .
---
1940 .
r orpho:cnet l.C sn ')s tn.nc s ol' orc:nn.
~
- -
.._ •
T"'>
D •
---
1.zer s in Anne lid d.ovc lo· :-- . . . n t .
J . E. l ., 85 .
Dentn 1iwl 106 .
Sc '-.1e op , ··1 •
1925 .
Die ~~1r c hung disDCrmcr .
Ener . !-~.rch . Znt . 1 ec} .
~;1er ,
1930 .
i~~ .
··ri1son , E . P·.
1904 .
Ex per imen t n 1 s tudics in t3crm1.nn 1
--------
1904.
Mosaic devc lor.:me n t in tho anne lid
egg. Sc ience 20 .
-------
1929 .
The develoomcnt of e~.:r: frr:t::.,monts
in AnneJids . Roux ' Ar ch. F . Entw .,
117 .
Ya tsu, N.
1904 .
Expcrimen ts on the dove lopment of
egc fragments of Cerebratulus .
Bi ol. Bl:tll . 6 .
-- ------
1910 .
Experiments on serm1.na 1 locrtlization in the err,g of Ce rcbrntulus .
J . Coll . Sci. Tokyo , 27.
1904 .
Experiments on tho localization of
devo lor:mcn ta 1 fnc tors 1n tho
Fomcrt1no og ., . J . Exp . Zool. , 1 .
-
Ze 1eny ,
~.
eh.
~d .
.?roduc t1.on of do 1.b1c c~ ln~yos
in Annelids and I.'tol1nsco . J . ~xp .
Zool . 57 .
locetlization . 1 . T!.1c r,"'rm re[!;ions
in t :Oe err,r; of Den ta 1il..un . 2 . Jt;x periPents on the clcavl:lr;c mosnic
1n Pa to lla rtnd Den ta lhm1. J . Exp .
'7. oo1., l.
CELL LINEAGE
----~
.:at•lrntiol", Fertilization a 1d Cleave e
in Crenidula
Due to the opacity of the living e .r;~ , the detail, of m.aturoti 1,
fusion of gem nuclei, and cleo.vat;e can best be ~tudied frora prepared
slides. The eggs are not sectioned but are mounted ,..hole. A lm··e
po 1er eyepiece and high pov·er objective are useful. Refer to Conklin
189?, Embryology of Crepidula.
Crepidula is a dioecious genus with the males fcy;er in number and
smaller than the adult f e1'1ales. The sperna tozoa r.:in.c:rle ·:i th the ova
before the egg capsules are forned vnthin th~ oviduct of th fe,al0.
The mature fe ales are sedentary, the nalcs locoDotive, and at th
breeding season, or perhaps once for all, the fn,Jales ar visited and
fertilized by these motile males. All the ova produced by one individual are laid at about the sar..e time.
Make a careful study of various stages in the two maturation
divisions (slid e A). In the same eggs find the sperm nucleus and
note its approach to the egg nucleus.
Study the 1st and 2nd cleavages, noting the direction of the axon
of the mitotic figures in the latter stage.
Details of the formation of the first three quartettes of micromeres and the cells derived from the 1st and 2nd quarettes can be
found on slide B. Find ewamples of the 8, 12, 16, 20, 24 and 25 cell
stages. (Slides A & B contain all stages up to 25 cells).
On slides C and D find stages in the development of the gastrula.
Note the blastopore, and the increase in number of endoden1 cells.
(Slides C & D contain most stages, through gastrulae).
With a heavy knife, loosen a Crepidula shell from its attachment
and find the egg capsules. Those that are small and light yellov.r
contain eggs in the eqrlier stages of development; the larger, deep
yellow or mud colored capsules contain older embryos and larvae.
Examine the stages removed and obtain samples from the other members
of the class for comparison.
A m_ethod of preparing whole mounts of
cleavage st~.§__oj'__q.z:_e_pj_dula.
1. Obtaining the material:
From the place of their attachment to the substrate , remove the
egg clusters to a Syracuse dish containing a small amount of water,
and free the eggs from their enveloping membranes vith dissecting
needles . Having freed the e£f,S , agitate them by gentle rotary
rinsing with a pipette in order to wash them and concentrate them in
the center of the dish . Change the water tvo or three times. Removo
the stripped capsules with a pipette, concentrate the eggs, take them
up into a pipette, and drop them, with the few drops of water in Whic~
they are suspended, into a vial 3/4 filled with Klcinenberg ' s picrosulphnric fixative. Fix the eggs for 15 minutes.
-2_ er; oYo the fi xa ti ve u ing a pipette of s all dian:c. tor e~uipped
• ·i ~h a syringe bulb, and fill the vial ··i th ?0 ; alcohol. 'l h in
70~J until the e .gs are 1hi te.
It is
vis ble to \roid ·; 'hin
o
long in ?0~, since the stain CEploy d is best ·~en it does not penetrate the :acro.cres . These latt e r should therefore b. loft li tly
P.cid . Thus the CGGS arc r P .ovcd fror ?o-j iunediatcly aft r th< 1 st
"rash which rcr.-.ovos no picric fror. then, hydrated in 50.-f, 35,~, anrl
,.·ashcd thoroushly in 2- 3 chan cs of ;·a tl~r .
II.
Staining :
After l.'ashing v:i th \':atcr, fill the vial with undiluted
haor:.alurn , o..nd stain for 5 - 10 ni.inutes . For the polnr b~dy stnr:c' ,
5 - 7 minutes is u
ly sufficient . After staininG , :msh tho ·o te;hly in
water , dehydrate , and clear in xylol . ROMOV the xylol used in
clearing and replace it with a small anount of thin dnmo.r .
III. .:ountin :
Cover glasses nust be supported . For this pnrposo it hc.s boo 1
convenient to usc paper squnrc·s the size of ? /8" cover glns<~ s. A
hole is punched in tho center of ench souare uith o paper pu ch .
In mounting the sQuares arc cl...,arcd in xylol , and fixed to the c ntcrs of slides b~r addint; three or four<ihrops of thj n damar befor e tho
evaporation of the xylol . Vlhcn tho paper mounts have dried , tho
cpgs arc rcr1ovcd fro, tho vial in \"hich tho arc storod by the usc of
a pipette drav'TI out to a l ong tap e r and hnvinp: a smc.ll dirunctcr at
its tip. Tho cgr,s nrc allowed to settle tov·ard the tip of the
pipette , e.nd one drop of the csg- damar suspc:-nsion is placed in the
c enter d epression of each paper mount . Tho damar is allowed to dry
to tho point of formction of a thin film in order thnt tho cr,r,s may
remain dispersed and rith the macromere quartette adjacent to tho
s li de V·hcn mounted . Ayrply thick damar to tho edro of tho paper
mount , iiTncrsc a #~ cover glass in x ylol and a~ply it to the slide
ov er tho pcpcr mount .
Alternative t: c thod .
The above te chnique v·as used successfully by Hr . John Yilford
in 1939 .
An ol der method sometimes yioldine; good r e sults is as
fo llows :
1 . F i x for 30-120 minutes in rayer ' s Picro- s ulphuric .
2 . Wash in 35 , 50 , 7 0'1 ~ alcohol. Leave in l nttor until yellow
c ol or ceases t o come out .
3 . 50 , 35;& alcohol , to '"ntcr (5 min ~ oaoh ).
4 . Stain in Conklin ' s hnomatox ylin ( 1 part Delafield ' s to 4-5
volumes dist . n1tcr , to ;•n:i.ch 1 drop of picro-sulphuric
fixative is added for cnch 10 cc . of the diluted stain . )
.. for 5 - 10 minutes.
5 . Wash in v~tcr , dehydrate 5 min . in cnch alcohol; 10 ~in . in
95% ; 2 changes of absolute alcohol; xyl ol .
6 . }.~ount in thick bnlsam with supported covor r;lnss .
- 3-
Coll - linong.
R~ror~
.c s
1. Generrl :
Euxle~· nnd DeBeer, 1934. I:lenent.s o.l Ex...
·i:"C'1tnl Er.br:r log:,
Chap. 5 . Carbridfe Univ . Pr~cn .
J.:cBride, ::. ·~ . 1914, Textbook of Er:brrolo ~·, Vol . 1 •
.:organ , T. E . 1934, Enbryology and Genetic n, Chap . 4 . , Col\.U''b . ,
Uni v. Press. Ne,' York.
Richards, A . 1031 . Putline of Conprlrative Dnbry 1 )mr, p . r::o
Wiley &. Sons, Neu Y0 rk
2. Special :
Cre _?idula. J. ~.. ornh
- . 10.
Arenicola , SternnpsiC'
. ArcL, f. Entwiclcl • .
Unio. J. · 0 rph . 10.
Adaptntion ~·r. F . Lpctures , 189 •
Eerl ,r Develo !l1en t of :nrine Annelids .
J. J~orph . 1~.
1932 Eorly Deve.ltopr:ent of Echiuroid Urechis .
Newby, 1.'!. \l.
Biol . Bull . ~ 387 .
1899 Development of Ischnochiton . G. Fisch r,
Heath , H.
Jena.
l\:eisenhe i r.1er , J. 1901 Dreissensia (Yollusc) . Zeit . f . wics .
Zool. ~ 1.
T1·eadvrc ll , 1•• L. 1901 Podarl<:e. J. }."orph . , 12._
1905 Physa . Z . F. wiss . Zool ., 83
VJ.L e rze j ski
1892 Nr:;reis . J. rorph . 6 .
YJLLson , E . B.
1878 Clopsine. Q. Jour.1:icr. Sci ., 18.
Vthi tman , C. 0 •
1903 Polyr;ordius. Arch . f. Entvrick . 2
.Toltere ck ,
1898 Cell-lineage. ,., . H . Lee tures, 1898.
'Wilson, "B. B .
J.nn . N.Y . Acad . Sci .__lb_
Conklin, E. G.
Child, C. 1:.
Lillie, F. R.
Lillie, F. R.
!-.'lead , ;. • D.
1897
1900
1895
1899
1897
--
o•l · the pi·· cttef' Q: he st::: .k d." 3!"-i s to
tnin ,am 'e[:.
ill res;.:lt i:. c .tRm r..::-tir -r ... h
of t~. l"l'till ...
""~
'tl-L: sp"r!f'i1. C? .a .
,..~.e soc-:
1 CJ~::.1
- · ' o f ~"'f':fS \":1.
·
· 1)t· r..t
t·l'e
,. "'
-~
·1.
!.... ;, r, _t of the :::'00Jt: , the st e k
ish of spermc\ to::
t f- 'C:.I l' •
C ~ "i' ·
1 (l t
.. t~~t Cf' te.~ ~e~: to avoi jn..,en.:.r..a i~g hen,·ily . Jr-r man: ~1-t'l'lnnt
cf' en cn. u~
o lys r'11: ·hi h res•llt s ~n abn 'Jrrt1 clea ·
m.'i. :ev lf'nPcl ~ tm.i..c e gs of erne fo ms de'J 0 lC'p r:o
th~l.' .
:11z .d eg~s . The_polynperm:c c:es of
1o~·eve ·,
o cleave .
... ...
ar~"lr>ss .. e ~s
· Use the r.1inar:· lo;~: po ·er of ~e compm:nd I ic.ro.,cq e
tr ~'o. ·i ­
:nRtPl:· 100 di~l.rJetArs mf'.gr. .i..fieA.ti c.Yl , Le., 10::\. fl~·f'-pi ~ ,.... an• l<.X
oljjective) frr Q. serv.r.t.)r.. ar.d study . This p rm t~ :>1Hi:1 n.i..1L ,~
Af,gs in a consh erab1e vchune of y.·ater in a Syrnc1..:·e 'R tch , Ja~.-;
t.h8t r.or,rrntrat.ion of Ren ':'l'ater by evap0ratic. it ! ot "npid .
~~,)P4phenomene can be F:een. readily ~i th this :ma . j ilea ic• ., •:h ch l ff c'•·d ~
rxc~1len t de fi n..i.ticr .
Hi0her po\"er'r. rr.a: be used i f nr.P. df'r\ ' £>, . ~
vb::erve ~per!'";atozoa ln c1e tqll <..mder A. C~"'V<"l'~L p. Aft.e-:.' ;;pet·m P"nt-·
tration .lr_ )erP.ir. has teen follo•·e ·: by obs~;n .tnG lhe egc~1 11 ~.!1 e
•:•ot<'h e;1nos for a
1e::>.:=:t :?5 1r.inutes, it ",• i:ll t>e inf'L:'n~t . vn t0 .tn~~ c·m lnA.te a secor.u bPtch 1 and per•ioct.icall; mou1t :;ome · 1' t.l1e8e un(' fl
~ c c..verrla s s to ob se.,.,vP ur::.der h..i. .h r pov er .
Zemem c ., that the y ~·c:-m2.Jr. ::.or:n8.l bu, n rhqrt time under these conn itlcn~ .
lffiHEIS :
Ob tain a fe~ w.fer llile~ c~Gs i~ sen wA.ter in a Syracune dl~h .
They nre approximat ely 1·~0 P'l:cr· ons in (lln.mct r as see!l f r )ffi above,
~nd 100 microns high in side view ,
Becau~e cf their ~hape , they tend
to orient on a flc..t rurface 'f'ith the animf\1 pole .c Lthcr abcv0 .or
.... elc"l, ."'E-lrely to the sifie.
t bser·re. the lRree immature rncleuf
(ef:rminal vesicle or nucleus C">f the primnry oocytP ), ancl the o:1
drcplets and yolk spher es in'the cytorlasm ~urrr1nding the nucleus .
Note also the thick cort0x of the egg .
Aft~r'becoming familia~ v·Jth the unfertilizP~ egr ,
b~r <"~. c'ld in.c a drop of frt·~ ~hly pr c> pared sperm ::uspemsj on .
tnseminate
r the CGf.'B
~. t t
at on c e t.y a circular P'lC'Veme t of the dish and observe chan;~r1 . Thf'f\
first changes wj_l l he gin a few seconds nft.er inscminat on • . At 21°C .,
t.ho follf'\"ing schedule avplies to 50/b of a populRt~0n of Pg("r (time
after fcrtili?.R. tion). 1st . polar body , 45 "1linutes; 2nd ~~olar body ,
60 ~inutes; lst cleava~e ~5 minutes ( ref . #2r ). The lsbcratory wil l
pro~cbly be warmer than 21°C ., ~o th~t ~Pvelopmen t will pro<' eod P'lore
rap i d1y . There are r.•any chan.c:;e R lr. the c: f_,G to obs~rve "t.1e fcr• e tht>
first p01Pr b(H'J.y forms .
Gh::.::f·rvc and r e cord the followi nG : l ab oaat N ··
t el"'\peratvrA , time of breakdo•trn of the r:rurminnl Vf's:i.clo , tiMr of flna 1
p or.etratlor of ::;r;0rn1 :'l"ad throur.l" "'1embr'l.r e , UmP of firf11' prle.r bod.
format ior. , tiMe of SPC ond pol:1r bod;: f 01·ma t 1or. 1 t il'10 rf' c l0nVf.1f;A .
The til:lC schednlo in the des criptTV(' text t'• lo•~: sl'Kl.l r' h~) lr1 A.! i. l'•• •i··
mstely if the eir tempor•at·1rc c1f tlw l::~lf.>l·ato.r .v 1e ulHJtll :: ~tv(; .
•
- rc. -
--
If ,t.:OSS i le, arra!'l;: e t c or s · rv'"' t ·o or ~ 01 e e gr_ ~- ;;• _c~. 2..1~
t cu.c::.ing im."'!~ edi£~ tely afte::' f e:~ t "l ize. t en . 2 - 0 min" t . c.ft.el'
fert l 1iza.tio!1 t:"ley w ~J J. e g i::1 tote ~n s b. d R J<~ rt by trans_tr :1
.~ el~- ~ - secrete& by the e gr:s e.x, ~ r~El to t~e , •.:_telli'1e, .,.,~1.-ren •
ny
2 0 i! i::J.utes , ~1e zcne of ie l'r 2.ro(1nd eac> .. ,.., :· .. ,. } · e as ·;ic:e os t.1e
e c: :·la:-:-e"..:er . 'l'he ~::crgi; (_ f ... t' c 'elly can ofte 1 b c . t de 0 :t - y ob s e:' v : ng su1er-n·'.:r.era.r v s ~J er2r. t '·ozoa anc. ot"1er 1r,rt i cl. s nt t ':e edge of
t .1e ~ell"r • 5 or 6 r.linute s sfter fert· 1ize.tic) tLe v tel l in e (fer ·
t 12.: z. ti~n) mem:~ra"1e '."i: _ ".:-e l").Oti ceable due to t.~--e fer · ti on of L1 ~
n , rrov: J:.·e r iviteJ.: ine s race 1:-0!'l jell._ eYtrusion . At 7 or 8 .inut ~,
-t:::..e er:tr&.noe cone '.;e_:: i!'ls vis ibl~y to f Tm. Fi 1d an eg: · owing a 1 rofL~e view of t':..e en tr[lnc e CC':1 e rnd t:~e sperm w'-'i ch ic- to enter .
For•t•
8 to 12 minute s or 1 onger , t:.:e s 1-erm is cles.rl r vi"' i.'. le u ts ide the
vi te:.. ine (f<?rtilL-:a ti0n} r.·. e:~br.:.ne at t ;1e ti p of the c ons p icuous entrD.n c e cone .
In t~'!e c o-..rse cf the next 8 to 10 minutf's, tho vitelline
rre!'''"r.:ne }S i::1dent oc sl:ig~"tl·~ :..:t its }.C':i.nt ot ccmtact vith t 11o en 'Cr2.nce cone . T:. ls tends tc .}' s cure t.'1e s l.C rm f rom vie •:; to s om ex ': e'!'ltJ a::1d 2. t 8.bout 2 C minut-. c~ e.:fter fert ' J ization t: .e eg r: \•;rin les 1
' ~e c .::-"·~· 1 -:. :- dJ.stortcd £1nd C' ':i te .: _r:rogulaP in sol.e,.·}.' rt amoe' o1.d fvs~:. ion .
T':.e e v: t~ a!ce cone "' as a lre2.c~7 f18tteJ.ed c cnsider~.' ' l'' , ':1t is st~11
,;,1· e; s .nt, rnd altl:.cu[h t)J.e s 1 en11 is 1,artiaJ.ly obscured from view , the
<;;,1 t:·'C1.DC t'l cf' its ~ ead into ~;,e cgr: is not co.cn}letod until orne time
l &t c r (ref . #12 , 15, 16) . ~)e final penetrat ion of the s1 rm. ead
tLru tr>c rrem1:; re..ne, J.eovin[. t".e n~iddle ri ece and t ai 1. outside , may
e.lso be o~-~ served . J.t a1'out 30 r.dnu.tes, the egc ronnds Ul ' ars.in , but
as t' · e t:J. me t:.lJ:-roaches for J s t :· olar body fol'IN· t ion , the e :-:·c ulo!"gr tos
i n a d. 'rr ec t ion· per pend icule.r to t' l l 1- olar axis.
If no ee;( s 1 ic so
t::at t::.e for min g :-OJ. ar l"lod·~· i s on t:;e l:. orizon , tho dish s ::.ould be
shal,:cen. The 1st }. Olar > ody !'.18.Y form at .< tbout 36 - 7:0 min·1tes , and i t
li e s in '~·\e s pa.ce beh!e'-'n t'\e oe;t:; G.n d the vitel line membrane . 'Ihis
sper.e i s wide r in the r egion ;;.f the ani;ua l ~ol e t~:ct n e ls e w>c r u • . The
seccDd }Ols.r b ody often for~~s at abou t 50 minu t e s and COli:..r~ dnly doc s
so ir:::.: 1' Gd5.& tely und e r the. f) r s'i:; .~ ola.r b ody, vJ·1ioh is thus J. 1f ted i nto
t~"lE.. 1 er i vite 11inc S fD. Ce.
f.:. 'c pe rha v s 80 Minut . s the eggs v11.l l begin
to C.iv:i.Cl. e into t •1 o una c;ual blc.~ t01~10 res.
Obs e rve 2"1d and 3rd clea v nr;c s
c.lso , i f tlme ye rmits. ':":1o 3rd div " sion, from 4 to 8 ccJ. l s, lJroducos
4 ni cr omc roo b~r s }--iral c1 ec. v e.[:{; (ref . #24 }.
v
1:-la c o some very r e cently fe rtilized eggs of 1Te r 0is in a drop of
fresh , thick chinese ink suS }:.·Gnsj_on (medo u p by ru1"ibiilg a .!.-te c o of
ink: on e. S y:re.cu ~ c disi1. rnois to n ed with s ca W8. tcr} in the ce n ter of a
Syracuse dish. As the ) el Jy i s socr e tea, th e e.tta c bed Si-'G rm caus es
8. c s.ne l to form in the secr e ted 5el!y into Vi'-- ich 1·articJ o s of ink
V!iJl I-<:mo trate. T11is is duo to inhibi t i on of jolly out:flow at tho
fOl..nt e f s po rm at tac hm~ nt. The ink thus marks the ontrG.rtc o p~:i. nt of
the s .~:c rm. Arter the: canal has fi l led with ink , add scfj. v1rt Gr and ;
if ti r:.c 1 crmits, obs e rve [.nd. record for n. numbor of cg[~E' the rc.lc.t ion
of t~;,u fir st cl E..a.Va go 1-l an c to tho po l ar b odies and tho 0nt roncc
~oj_·r1t rf tho S.t erm as marked ':J~r tl~c i nk (ref. hLl2 , 1( 1 19 ! .
(C aution:
do not le&vo t bc .i.- ieee of cl":inc sc ink i,n t . dis~ of s ua water; lt wilJ
dis i ntcgrr. t o } •
NEREIS ~
Exo.ggora ted En tra~~..£ Cone s: ·
flace some Ncrois ogg3 insomin~ tod 5 to 8 minutes oarlicr in a
Syracus e watch glasu co nt ~:.in::.ng olkc:.linc 1~a Cl (pH 10.3-10 .5). Ob§'lerv~ i mme diately.
Tho vitC;ll ino membranes will elo v f'. to duo toa
sudden tnh:ibi t ion· c£ jelly ro 1oas c t h rough tho l':omb rnnc E.md o. subs o uc.nt accumulation of tho ~ olly in the 1-o rivitollinc.. space {'l"cf .#9).
-3he vi te ~l ... nc :-:cm""rant re11:a i!.s permea · le
f'nt r s
perivitelline spnce ns the jelly swell~~
nf
ruern·
.,_.,r,ne st.::-etches out tho sporz:1 entrance
ne .,_.,et ''08. n rn rnLe a d
e[jr- surface, for min a lone, filarr.ent which fro .1en 1: cnusc s rna rkod
it.· ".':1t.., ti o. of the menbrane. If the cgt:," h""·.Je b')On }-.cpt in nn ccb . ~h ~ y rn~y b~ccme polyspermic upon insemination a.d sh v ~umerouR
c; ... 'f'F!"i"a .ed entrance cones upon treatme. t ~·ith alkalln, JnCl.
J bo l
ccn minutes after treatMent the sperm head rnE\7f b0 seen movin . o.rr of ~
the perivitelline space to fuse vrith th<' ft[".g surface, at thi !1 t rtc
thn membrane inclenta t i(m is relaxed .
If tho s eg s are are full:
r emoved from tl1o alknl nr .trac 1 to sea \''a tor 1 nnd washed, somr :i ll
develop normnlly v·ithj"!"J. the rPised membrn.nP-s . If loft in allm~ n c>
·;ac 1 the opt imun h~ ngt.h of time be for<" wa shinp:, and if th nlk~lin e
n ac 1 has been chn.n~ed once or twice to remove most of the co a 'i'a t ~;:r ,
tht> eggs may bt, completely freed of the lr m cmbran . R· T!:'le se "n::~J;:c•c' ''
<'gr, s have been tsed for experiments on the duvolopment of isol·1t d
~ lacto~eres (ref . #8).
tiere is:
Cer:trifuged Eggs :
If time permits, c c:n trifuge some unfertilized Nereis eg~s in t.lt.
:: morson electric cer.trifuge (cover off) for 60 minute' s . A l:1.yor of
0 . 95 molar sucrose (in distilled water ) at the bottom of the c ntrifuge tubes prevents injury to the eggs . This is somewhat hyp -: rtonic;
but provides an adequate support for the Ncreis OBBS without lnjurin l
the~ in any way .
This amount of centrifuging sopn.rat~s the vn~iou
:l' crmed c ornpor.en t s of the ee,g into several strR ta ( r c. f . #7 . ) .
Insom··
1nate the centrifuged eggs after washing off tho sucrose ~ith sea
v: "'. tcr , and observe asyrr'Jtwtrica.l jelly- extrusion .
Is more jolly ex··
trudod at tho cc ntrip~ t al or c entrifugal pole?
I f Cfj(!;s r;t r a t ified in an u l trn.centrifuc;e arc nvailnble, compare
~hc sc with the Ne rei s oe~s c entrifuged in the ~merson Glectric cen·,, rifuge at about 10 ,000 x gravity (re f . # 1 0) .
£I r ee ding
ha bl t s of Ner f:: is _limbo. ta
Nerois swnrm in Eel Pond n.bout an hour nfter clark at c ~~ rta.1
phn.so s of the lunar cycle .
(Sec Lilli~ and Just , 1913 ).
Cn each
two app ropr iate e v eningf; , about nn hour n f ter sunset , half of th t.
cln s s wi ll r,a ther on th ~ flon. t lnr; stagQ behind the Supply r,f;pnr trr.~. ; ·
to ohse r v8 t his interesting phenomenon .
vJ:I~ET OPTERUS
:
The c hac t optcrus <'[jg is rather darl: and rrrmular .
It is sli gm.:1
more than 100 mi crons in diameter , hut ticfore ftrtilizntion the egg~
a re of ten not quito spheri c al . The p ~ ln.r b odies RrL larger than in
NF reis , ~nd thG egg divides to f orm two unequnl culls by mc nns of a
polar lobe at the vege t al pole (ref . ~14 1 18) .
Whon the egg is t aken f ron the femAle it containe a large
nuclou1:1 (er-rm i nal vesic l e:), as docs the N(:reis egf , but un like tho egc, of Ncrois , i t cpontan v otH.:ly undcreocs partinl maturn.t1.on \"hen p l a c ed in sea wator , oven if not fc:rtilizcd . A number of
species of e gr, s pc.r tl y ma t u r e when t h::y vntor sea wntor and PA.stoel.
( ro f. #21 ) has sh own tha t t h :.i s i s dependen t upon the presence of
Calcium in the s oa water .
~mmature
-4Cl-wetooter s e ::s ne ·elo ~ 1i te :>".. ldl: . Ii' e. :~s
just after t 1·e .rtial .. atu:>ctl.:l in sea v~·ter n' 1 ce.1 C'"l! :let d ,
t...,e. d~·e:o-c as rz idly r:.s e rs insenL . .ated 12 -J S r'1'1'tes _'1rlic '
..,._~en fir"t ~laced in sea ;··~ter (ref . {1:23) .
t:i.:~'e
mted fr r.1 fer tiJization) : J,st. ol;>r body 14.5 i:-!l.nutes ; 2n1 o l'r _od, 1 27 11i.1'1tcs;
_'2 e::>.r s'h.-.pe(~ 11 stz:_,e , 4t. .::.ir:t te:.q polar lote 'llr; , 52 1 :nut s ;
:~:e"'v;'l,e .. ,it'-. _olnr Jo~·e att"!c_ed , 58 :i::1utes~ co; ·•lct ~ cl..:"'·nrr
··:7_t., ooJPr J.o1;e resor'!Jed into l")ne blnstomere , 62 t~'ll1 tc~; 4 c 11
s t:> r.c ~ 22 ,.,~_n t2 s . If t_ e lc ~ or r tory air ten:!.er'l ture is D L 1u t '1 ! t?C.
t'-10 'ievelop!'!en t ;·;il1 'ne moY'e rapid , :>nd about at t e rnte i~ldiccttel.i
,,..., t'-.e desc~'irtive text belen .
"'ten t'-.e e~:s "!Y'e t"\:::en from t ·, e fcdlle, t>t.: l·.r-:e iM -:tnre
:•nrle, s is in t 11e ce ·1ter of the e ,o: . ' fter al~out 1r.: "'Flutes :..n sen
·,"3te?: , r;.e.t1.r::::tion viill '''"'Ve proceeded to the :_,etr\p~~se of tl.e first
,,ol~r· snindle ,.,t ·:~'lC _ st'ln:c ~.eve loorent 1.s ~r·ested unless the i..;
·ls lrl3er·ir:' ted o·~ Artifl. c1..:> llv activo. ted. T!~.e s1. indle c~nnot 1
_tjs tJ.nguis1.ed ~s snell in the livine; e r; , but it rnll ~e o1v·ervcd tnt
'-:~A rel"tiv:l;t clear. ref):on of t~e nucleus nnd S\1iNlle 1.s now Jo ..
02 tod T'i t,.., eccent rlC" lly .
It rer-c\~es tbe surf::cc of the o·~r:; in t 1e
'; ol"t re;.ion , ''i.1ere t•·e ~10 l ~1r bodies \'lill be ~i ren off .
~
T:f e_?:?;S Pore n0''! lig>tly inse:;inR. ted and st1.rred,
Q few snerm
··:it.,in 5 or 6 ·.
~lr.t..tcs , t 1 .e 'r1teL\.1ne nembrRne ma/ 1le seen to sepnrntc f.,om tl1e eG~
:nlrf ['r;.--~ , ')fte:r ... ~ic~ t1rr:,.~ 1.t is c alled t'"'e fertilization r,cmbrr.ne .
It 1s not C" YJ.Spl CU01S Pnd does not elevo.te nuc~1 nl,ove t''C e~r:; surf'nce .
!';; 10 or 12 r1.,.,,, tes, tloe egc;s , ·ul-ti c 1~ 'rl')c~. becore 0lmos t spher1.cal
0f ter fert1l1.Z" .j.ion , . ~re seen to elonn;"te 1.n nn nxis perpendH'11lnr
to t'·e ''OJ. 'r 2XiS .
r 1l S iS p:rep8rR tor,r to formation of th
1st pel~'
1·•ody 2 t ·'hout 12 minutes . In this d 1 rision t he egr:, thus ns sumos
nppr oxinF tely t r-:e sha pc of r~ bla s tomer c , a lthotl.gll t11e polar boc1 j s
r vcsti,q;inl cell.
~f t cr tl1e lst p o Jc:~r t) ody for•:1s , the Cf!.B ngail'l.
:o1.cnds up ( 16-19 ·lt' inu tes) 1 1m t it c lonc;a tes again in the S(.~ne nanner
to form tre 2nd polc.r hody a t perJ,aps 23 minu t es .
The 2nd oo l Qr
'1ody often forms under t'' e fi rst , '.'(hi c h is thus r-u shed avTny'f rol.il the
urface of t~e eg~ . The e~G R~ain round s up (26~30 minutes ); The
e?.'g pronucleus m2y n ow someti:.'e s be se e n rnigra tin.c; towrrd t'·e center
of t he e c"rs , Pnd occ asi ona lly t he sperm pronucleus mRy r,lso l)e de tect ctl .
·().y 35 m1nu t es , t he c J.enr z one np, s extended from t~e polnr region to ''!.<> rd. the eiuator of t!10 e~('; , HnrJ "tt 37 - 40 m~nu t es c
ty5nc o. l ' pern·
srap<_:J cJ 11 stare is re ~\ c '··ed . 'I'}-le pol:>r bodies li e a t n ~Josit1.on cor·rcs pondin~ to ~here t he stem a tta c hes to a pear , and t he bul~e whic~
forms t he po l r. r lobe begins quite suddenly a t t'~c anti - polnr Gnd of
t ~-e.. ego_: (40 - 43 minutes ).
··rJJen t i-.e ~'o ln r lo':>e is fully devel oped ,
"o'''evcr , it corresponds to t l1o stem en(! of t:re 11 pet~r 11 , and t he polnr
bodies are o ~osite .
rr~.y
b::: :::;een adl~erinp; to t~c egrss almost int'1cd iately.
0 1.'se rve t'-.e extr11sion of t 1e nolar lobe y.,~.. ich cont·:ins conrse
eJohnl?r m" terial. ')~r 47-48 rj nntcs t '•e cJePvflgc furrow 1)eJ3l.ns a t
t' •e an1m8l pole n nd p1sses to one side of the polar loho , wh1.ch
tl1us c oDes to )e A.tt" ched to one of tlle two 1'Jqstomercs . T}1e resor')tion of t "lJe nolAr lobe into t"lJis lil<'ls torJGre cFluses 1.t to becor1e l'lr~c·,•
than its m"' te 1 2 n.d. !l t a hou t 51 "":' nu tc S t··• 0 smoo t ' 1 UnO lU8 1 blP. st om rc S
lie ap:ainst ePc]-; ot'.,er . ?o l:;sperm1c ee; .·s VJill nov; often be in an
Rbnormal 3 cell s ta~e· .
r:.7,r 60 m1.nutes tl.. e t•·•o 1 "~l8stomeres are quito
fused togethr;r . ' t 67- 70 m1.nutes t'"e 2nd clcava(J'e takes pln.ce . T!1e
large blastomere again for~s a
lobe , nnd n 4 cell stage results
h
n·
t rec
Q.,
90 minutes or
-
0
'i
J
r'
';J
0
J
1-
0
r'
ct
.;,
(I)
~
Q
1-i
0
(fJ
..
0
----~~----- ~-------
-._ 1;.·:cl.ei in t~e 4 cells ~ a:· rc· 'ily )0.
o1.t.
3rJ ch ;ision ta <e"' pl~ce 1 fo~·~ -'..t ~- 4
•
. ·r ·2 Irlcro,...,e:r(..:; .
ryroflle '':!..CYI 'Jill rc • al t' \..: !'1~~-.!1' 1.. m•es, . lc::.•o _,...,.es; c !d !JO~'--- 1 o .:!.es .
~·.
)olnr. ie·J vnll sho·1 tl1e ·ctnt~ l dis)~~ce:--:c·r:t r:>.;.n t· e ~.i~:."'·''"'Orcs, j'CS'lltins fro!"" spirnl clc. , . . ~e , r ltl m "h
:: --e d.ispj_:'~~: .. t::!~ )_s r.ot
~e t or conspictlous it1 Chncto1 tci' 1~ .
·~r-r1ier ,
t·~e
ch~·>r
, 9::--S7 mi:: ttes t
r~r':
.. c
;: . ,.,
·....,I~·
-----
fl"e;C · s:-..,11 s''::'-lc-• troD of 11 •i•· 11 sncr.r>to 7 0R d1rectJ.r fror.1
;~10 t::stis 0!:. a SlldG .
.· C sner:n r:nst'' not' e dilntcd \'Jit' fll\'r "CC
':ntcr ;·:·-·o tsoever . Se:1 urc~il · snerm~to~of-l are r,oo l fo'" tris lcc<U0"
• +; 1s especiC'. lly ensy to o1 t--:J.~1 t o:n 11 C'.ry 11
J.rt•ctly f' O" t' <.) tcstJ.. •
:?:'" c e ~ ''!'op of S-32 r: te·· '1C".rbv on t 1·c slide o t 1., t 1 t 1 os not
~ouc'' t 1·e dro-o of sperm-" tozo8 . ~ ..:1 t:O or'.Jin:trv Jo·· uon l' {n11nro:~.
-~-~--:;-·) cxa:njne. tJ...e s~e:;:m'tozo"' ~nd note t c ·-~C ..:,l'Gc of "et'<lt: cs0CCii'.}J.~r 2 t t e c ,.,.e of t',e clro1) --·her•c t \·ov rr.~· ;1c sce}"l JoOl'C Pcndilv.
fro:: to::kb n SP'lall f3l"'SS r- otJ , or~- ~·tch sLicl- ,
ot'C' ol~Jcct rtnd "
dt"'~ 1 ts tip fl_"'O!"'l t\-,8 d.-rop of ser. r: t t2r i tto t.•c drop of s~ c mn to zot'.
so that "' connectinr- ';::.'idge is es t hlis• cd .
s t '·e s:)e •m diffu se
into tl;e SO:-' 'Nntcr .'1 ~!'f1 1 ·iont of C0 11CCntr8.t ion is cst[.\1~lis:"cd .
rotC'
s•·Jilllr1inp; octi,rity in rcl2.tion to conccntrotJ.on.
or
Tf Cu.rnin~ ia
e~::s
ere nv~ iln blc , o 1)serve t~!C r·igr·1tJon nnrl fu"'jon
· 'h1le Ct'!.Inin[':ia e·-.'':S arc; small ( ~1bont 60 ''licrOJ s 1'1
r"i_,,r,e'Lcr ) , t 1·ey :.:n'e cle[\r ~~nd s>1o·:, t:·-.c pronuclei in the ll.vinr; stL".t.c
csp•'C'.L.l ly 'Jell .
'I'l-!c cytoplo.s:-1i c const1tuents of t !•Csl~ o~~s are very
l'Cf.lci.i !.y stra tificd into four zones in tl1o ccntr 1fuge.
ilf t'·v! pronucleJ .
Durin~ spf:\rc; time s tu dc n t s m8 y nnl~cr t'1 ?(e s 9ec ia 1 projects re lot inr; to fertilizr. tion or c ell - lineage . T' e follo·1i n'3 prol)lems nrc sn :;r;ested , 11ut stn ~e nts may forr;n l ~ tc t ',cir oan p!'OJC Cts subject to t 1'c
11 npr ovP. 1 of t'"'e ins true tor .
Pla tynercis from t)~e
ConCon pr·re your o1;sorv"' t1ons '.' it l-. t bose on h=rei s liml;n ta .
1.
FertiJ iry; 8 tion of Pln t vnereis .
Colle ct
Cf.l:rn.dett ;;:----~'::tr f 1 f1o[•' tfng s t nr;e" 'l t t' •e rio;ht ph::>.sc of t 'he· J'Oon .
sult E . E . Just s pnncrs on hreechnp; :.'f1 1'i ts ;!nd fort1lizo.t:Lon .
2.
Develorx:1cnt of Isol.~ t cd '8l2.stome res of Norois . r.~e111 ove
l:'eml)rP.nc s of i nscr·nna Tiel rlerc1 s egn;s •1Y~~~0aT!ca l1ne lTnC 1 me thod
pY'oviously '.~l::-~s~ri1)ed .
Usc serri - sturilc tcc ',nioue Vlit', nll dish·•s
!'lnd instruments . 1 a 1re Snernann !"less needles af t er dlr<.ct:iono of
TTOl_"'St -diu s in J cClune's r:1i CY' OSCO:-'iCRl Te:c'nir~ue . Usc srr.n11 vtr•tch
~Jasscs (pi'cfcra 1 ly Cohun 1)i[\ ) ·:nt'l 2. t~'ln lrycr of f1lter-ed q~::tr
mode up in soc ''Jr> tcr , · s opcr~ t1ng fmd 1 sol;' t::. on eli s~~c s . Sc po ra to
t' ~e lJl~,st omeres v1it!, the f1ne tips of t' c r;l ss noodles immechatcly
eftcr the cle•vf't;e fu rr o·:r is comulc te . 'rr.•ns fer to scp:•rotC' 1~:oJ· tJ()tt
disl'es wi th :"'out1~ "lpettc ,' nd pc:r'"lt 1.solctc<l l')l' stomcros to clcvol op
in cool :"lOist c 'Y:!1ber s . Observe nt fro;u8nt 1ntc:rvals .
T.)e veloumo n t of Centrifu.c;,ecl l!crcis ~~n:s . Ccntr1fu~o lorois
3.
cr:;gs for 60 to 9o n iniltes-in Eme;-son c lcct r1 c ccntrifutze witll cover
-6~us cri aa d.
,2.e~.v-~go .
·'o ::::~ :~ng r r
~esh off sucrose in sea water, ins .in~te and study
S.::at i stics as to the number of AB E\nd CD blc t m~.-rcs
o:;., C(.!J. tr l pctal or centrifugal ends of the contrifu. cd og. ,s
·;ou ld bt o f int , r ost .
Positic1;. of microm6tes may "lso be noted ln
·~: lat ~or: to ~t.r·~ti.f i cntlon arrl in relrtion to . gg 1olr.rity •
•
- 7Fert i liza tio'IJ.
s i "..•lio~r~, phy
~enersl
1.
2.
q eferences!
:rray , J ., 1931 A te:-dbool{ of .::::!1eri.nentr,l yt lo~y . C"'-:Jl ::.~ictsc
Uni v. Press
Just , ~ . E . 1939 T~:e li olo:_)y of t ~'c cell surfnce . L.l:;ldston
P~ila .
3.
4.
5.
Lillie , !:" . R . [tnd .E . E . Just 192t1 Cl1nnter 8 in Co·:; 'V,
Genernl C;.Ttology , univ . Chict.~o ?re ..s •
T. F . 1927 E:cperimentnl ::i~')ryology . Coln1 ~:ie
Un i v . Press .
"Ji ls on , "'3 . B . 1925 Tl,e cell in de~Telopmcn t '1nd ..1crcdity .
t
::<'tU'ther references relr tinc; to
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
19.
20.
l~ooratory
r:ork:
Costello, D. P . 193 9 The voltunes occ u.pied by the forr:1cd
cytopl ~ smic co~p onents in marine eg~s .
Physiol . -ool. 12, 13
Costello, D. P. 1940 The cell orT1in
Nereis limba t1~ , 3 iol. ?.ull. 79, 369 .
CosteJlo, D. P . a nd ~ • . • Younc 1939 The mcc0nnisn of membrRnc elevation in t he err•,. of Nereis lJ.mbatu
C o 11e c t i ng 1~ e t , 14 , 210 Costello, D. P . 1940. Tl1e fe r tiliz a~) ility of nucleated and
non-nucleo ted fr . '~ents of cent rifuged
Nereis eggs .
J • l~ or ph • , 6 6 , 9 9
Hoadley , L . 1934 Puls::>.tions 1n t~e Icrci s e3~ · Pio l, t:3ull .
vol. 67, pp . 484-493
Just, E. E . 1912 The r'GI'ltion of t he fJ.rst cler,vage plano
to the entrance Do int of the snerM . Biol .
Bu 11 • 2 2 , 2 3 9 • ..
·
Just, E. E. 1930 The present status of the fertilizin theory
of fertilization.
Pr otopla sma, 10 , 300.
Lillie , F . H . 1906 Observations and experimGnts concerning
the elementary phenomena of embryonic
deve lopnen t in Chae topterus. J. Exp .
Zool., 3, 153.
Lillie, F. R. 1911 S~ldies of fertilization in Nercis
J. Morph., 22, 361.
l·illie, F. R. 1912 Stud~cs oTferti1ization of Nereis,
III, IV, J. Exp. Zoo1. 12, 413
I.illie, F. R. 1914 Studies of Fertilization VI J. Exp. Zool ..
16, 523.
}~ead , A. D . 1897 'TEe e arly developnent of mar~ne a nnelids.
J. Morph ., 13, 271 .
Morgan, T. H. and. Albert Tyler 1930 The point of entrance
of the spermatozoon in rcl~ tion to the
orienta tion of the em~ryo in cg~s with
spiral cle a vaee. Biol . Bull. 58, 59.
Novikoff, A . B. 1939 C '.~a nee s a t the StU' fac e of Nereis lir 1 ha. tn
egf,s after inscrnna tion.
Jour:-1'!Xj). B::.OT:Vol. 16, 403-8 .
Pastcels , J . 1935 RccDerches sur le de t crminismc de l 1 cntr ee
onm3 tur a tion de l'oeuf c 11e z divers invcrte-
8.
·.'hital~er ,
J: .
1931 On the rate of oxy~~~~ consunJtion y
fer tilized a nd unfert i li~e ~ e:.. gs. III 1!e1•eis,
J . Gen. Ph~siol
., 15 , 191 .
~
23. ··'hit al·:er, D • . : . 1933 ibi c~ I\' . C:k'1etopterto, J. Gen.
PhysiolOGY • 16 , 475 .
24 . ··· ilson, E . B ., 1892, Cell Tine:; Ge of Nercis . J. ! or ph ~, _§,,
361 .
22 .
D.
-
Lttn>tr Periodici.!l
1.
2.
3.
4.
5.
6.
7.
8.
9.
r. .
F . and·: . 1T . Hess , 1940 ( 1\.t l ant~ c Palolo)
Pu~li c ation ~o . 524, p . 21 v nrnes ie In~t . of
·'.!a s !'-!in"- t on •
F ox, 1924 Pr oc. rt oy . -S oc. London . 3 .
HemDelmann , t'r ., 1911 a~ereis d1.Uner ilii) Zoolor· ic a , Bd . 25 .
Izuka, .4. . , 1903 ( J :1 pr nese P,l. lolo) J . G oll.. Sci. Imp . ·u.•ToJ<yo 17.
Just, :::;; • E. , 1914 ( Pwtynere is me~~rt lops) Biol . Sull. , 27 ,
201 .
Lillie, F. R . nnd E . E . Just, 1913 ( Nereis li mbn t a )
Biol. Bull. 24, 147.
Mnyer, L . G . 1908 ( '·_ tla nticPalolo \(Jorm) Public~tion 102Carne~ ie Institution of ·~Asb in~ton.
Sc ott 1 J . '.i . 1909 ( l~mohi t:cite ) Biol .. Bu ll. 17.
·;: oodworth, 11 • I1 c. 1907 (Po. cific Pi'1.lolo) .i:iu lT. ~·· us. Camp .
7..o ol. 5 1.
Cl?,rk,
..
•
•
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..
..
•
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•
•
•
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•
•
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..
•