F.cdty ols.i``!.r. Aiid Agriclhm Urtv.nlt Rr.lplrd{ PL!t

CENETIC IJIALYSIII OF TUMAN Y CEROMOSOME IN
PAKIIiTINI POPI'IIITIONS AND IMERTILE MALES
r,ry
?
SAIITIA SIDDIQI
01-rdd-!583
D.p.rtD.!r
of
F.cdty
Bhcldltry
ols.i''!.r.
nr M.lr Alt SL.t
Aiid Agriclhm Urtv.nlt Rr.lplrd{ P.L!t
20rlE
!
cn0[cAnALYTTEotft rNrcEoxxrD|lll{trll3TA!|r
!@T'LAIIOIIE AI{I'
INIIB![,8
XAI,f,E
w
EanAt|DoNQ[
U{AIIE
Alb&ibdilF
ftblh.dd
li...qtddft.ftefFd
D..-..CE Ib
h
Ht
D.ttri-adn {-!tt
A.adg
olsdd.{
tlM.lr AI ELI
Arld Atrhlla u.hr.dt rr.!ri& M
2at
ii
CERTIFICATION
c.ni6.d thlt thc @.tctrts dd fom of thab cnrid.d coc.ic A!.t b dfg{Ds
Y Cnrcdotod. ir P.li..r Popul.riolr d htinil€ M.16" sbnitcd by M!.
SrtEr 8iddiqi h.v. b.m foud sthf.cto.y for
rhc
Fqui@cnt ofth. d.g@.
dr,t^ l1a^u".u
(Ih. Alika Ma@ot
al
-f@'.
V3n t''a 4-'
A^xl.ue, -
(Dr.
s't1
Arid
Agric'
0/
/il.
Arid Agricultu! Uniw6iry,
e
Y@i! MB.dd.q)
Univ.6iry, Rr*dpiodi.
at/',n
yl^,$----+<
Arid
Agricute UdGity,
aettattttoqOntfr a! slstlrrt,ttufu,4
thtuadrrt0tw
CONTENIS
LIST OF 1ABLES
LIST OF TIGURES
IIST
OF
xii
AABREVIATIONS
ACKNOVLEDGEMENTS
ABS'iRACT
I.
INIRODUCIION
I
I.I
I
YCHROMOSOME
l.l.l
l.ll
llsdtiao! o! &ldic
1.1.3
l0d.ad.Iit poly@obi@
3
l.l..1 rifninLllit . Pbltrorpti@
1
Silglc nu.hotid. FlyDoahi@s
2
(SMt
12 ROLE Or Y CIIROMOSOME IN P,oPULATION OENErrcs
2
5
1.2.1 nisto.r ofP*irrrli popui,atid
6
I,3 ROLB OT Y CHROMOSOME IN MA[.E INFERTILITY
l0
l0
lt
2,
RE}'IEW OF LTTEMTTJRE
t6
2I
l6
PROPERTIES OF THE Y CIIROMOSOME
2.1.1 Y Spoifc Micrg.Lllite
t7
2.!.2
l8
Y sp@iEc
Mirilt
llites
IE
2.1.4 SirgL NBlcoli& Polyoorphio3
2l
2.2 MCROSA'TELLTT? MUIATION RAIE
2.2.1 lrfios.a.llib M|'rlid Mod.l5
L
19
22
2.3 ROLE OF Y IN INFERTILITY
25
MATERIAL AND METHODS
34
3.I
IDENIIFICAIION AND CTIAMCIBRIZ1TION OF
34
NEV Y MICROS,{'TELLIIES MARKERS
3.1.1 lddnifiord@Of NqMicro€d.llit
l4
3.1,2 [email protected]
Mic.!.Llitr i! Llddoi PoF!.ri@
3.l.at Mdlipl4 rcR
35
Andyri!
39
ESTII/IAIION OF THE MIIIATION RAIE IN CROVINO
l9
3.1,2.2 Stalirticrl
CBLL LINES
3r.l
SepL Col.cliod
39
3.2.2 EBv Ttudmati@
39
3.2.3 Cc[ CqDtilg
40
3,2.4 DNAPEFEIi@
4l
3.2t.1 DNA
3.2.4.2
rtld{ion ton Blood
DNA.xtn
li@tln
42
lrs$for.d Lrdphoclrct
32.4.3 Ederd@ ofDNA
32.5 SiatLpl.trcR fcsli!!.sc DcE tid
32.6 Cll Ebclr.phdsi3
3.2.7 St tirii.d Arabli!
3.1
45
MALE INFERTIUTY
45
3.3.1
3.3.2
46
Suplccon.lid
Sce@ils ofAZF !.gi6E for deLti@
3.3.3
49
3J.3.1
ARII4S
ICR
5l
PCR{,FLP
3.1.1.3
4
RESTJLTS
53
A}ID DIIiC'T.'SSION
54
4.I IDENMTCATION AND CIIARACIERIZ{IION OF
54
NEV Y MICROSATILLIIES MARKERS
4.2: CAIrcutr[lON
OF
Mt TATIoN RA'I!
TRANSFORMED CELL LINES
viii
IN
EBV
63
4,3 MALE INIERTILtrTY
4.3-l S.l@i!s
69
of
Aa agi@
foi
&lclioB
69
7l
4.3,3 H!'logFupilg
4.3.4 ScEcrias of CDYIa^ FlrroryhisE
5.
7
78
SUMMARY
&
LTTEMTURE CITED
E5
AI'PENIDICES
lt0
T.bl.No.
T.bl. 3,1:
YSIR
t bL3.2:
Trblc slo*ins
rtbh
33r
T.b1.3.4:
t!8d
Da*a .nd rh.t sqtuq, le.l
Ddr4 d6iglared B Ys IR4.
It. disribdid
of
dis$.d @lditid
M!.k6
a.l.lr
38
tu grouF
bsd oa
rt6
da! tb. @6pld. oi peri.l deledoN of U. AZF.,
&d c esioN
to
Biallclic
Edt$
along
$.
m.rhod
ro d€6n€ haplogoups.
r.bL
spl6
th6c
coltaililg .ll
u.d for ihct Vping
47
a8
Mld in
58
rt@i'lovcnll
6t)
H.t bzrgGiti.s &d nubd of AIei6 for atl fivc
Pllir|lli
b
crhlic ercup3
TlbLa.ll:
H.te.@'g6ili* .!d clt
!atisstr
Tdb4.r3:
HaploqF c@birniod &d lh.ir
rFpulatioN.
lttc
for all fire
n
&!qwy i! P.tkidi
6l
populalioN
T.bL 4.3.l: Subje& with or without d.l.tioB i! ditr@rr haplogbupd
Trbb aSrt Eaplo8m+s 'sig'-.at in tb. bfarile Fluhd@ h.vi!8 vsio6
76
TlbLa33: Showingib.resultofAzF,bddccgioB
79
rrbk 4.r.1:
Haplotype
rLtozygolity v.le! io Pakistali
d.sE ofdi.ce
62
12
No. Titlc
P.g.!
rig l.l
Loc{i@ of wioG.thric 8rcqps i! h&ietan
9
Fig 1,2: Cytogtrlic fatuc of tn. Y cbM@nc slowilr th.
t2
.pt@xilrl€ l6.rio6 ofAZFa, AZft ed AZF. iegioD.
Fis }l
M.? of P.Listlr 3h*ing thc ld'iotr or rb. v&iou .tt!i.
t7
u!€<l
in
|}e
$dy
90146
Fl€ 3.2: Phyloga.tic te b!&d on rb. Y chrcn@Dd bi-!.ll.tic 50
Fig4J.: Rcpeat uit A.{eey ofDYS648 ldu ir P,tisra!
55
Fig .l.lbr Rep€at dit tEqErcy of DYS549 locu in Patisiaa
55
rir 4,ls Repear bir iEqqcy ofDYS6SolocN in P*&ia
56
Flgdld: R.Fduir t queyofDYs65l l@u in Palisrar
56
Frg4f.: R.pat uit A.q@y of DYS652 hcN i! Pr&iiti!
57
rE ar'
oYsoso nnlrio6 ots.*.d in EBv !|s&@c<t 65
Fi&
Itrphoblai.id
eI li!B.
Sbowi.g
rl. fi"gD@t
sizc
b.s plir
4h Fat dd [email protected] of eFlt @it itr p@ hdi!.
DNA tuplc pFp.rcd fiod bl@d (B), &oo ttdfm.d
Itrphdttes (LP), Aon rafod.d lFpho.trca .A.r l
bclow
oonrhs for srewu
Fi8 4 2
2r
Gir)
DYs652 duraiion obscrycd i! EBv i6sfom.d
Ituphoblastoid ell lirB rftd 90 g@iatios. ShowilS th.
tlsb.nt si4 ha$ pai! bclow ecb F.k md Mbc. of rp.rr
@it ir F@tlEsis. DNA
66
rupl.
pFprFd &!@ blood (B),
ton rrafm.d lFpheyr.s (LP), aod rl@[email protected]
IrDphar*.n.3bo h5 ror 8D*rh oiP)
Fig,t23: S&difeE
Fig
43.1:
lraphoblasloid c.ll ties
tid.. Cnph
of ells ailq .v.ry
v@ ucd
forthc
calculation of doubling
showing the Ctos,lh
rcds oI Mbd
24
2%
Asmse gel showing
rhe Esults
68
i!
lN
ofMultipl*
PCR
for
73
AZrt' regiotr
Fi8a3:: 2%Ag@*gelstowilgth.BulbofMultiplqPcRfor
7)
AZFo
Fsic
&{3a: 2%Arrstddori!s6.rqli[.di.nof010E{01085. ?5
Ib rqL rtu qlltldd !! iori{ ni(loddrddt
h&AAc nCtod
llta3,a€ Pvt tr dir-tuc)nrdbr{DqtLE,3 t-&h
ll
.dqLdorirFs
oftle
dt
CDYtr ad b.
EBV
ri{€P12
hfb!
lBE
Tri! Bod. EDTA
s13
Soditlh Tro EDTA
NH{CL
Kgc03
MAK
A.BI
MCI
tTs
iM
ACD
TET
HBX
Floe*al
lrbeLd
t
Do*
FAM
TAMf,A
[email protected]&tt x)'rtodEi!.
DMSO
GgNDIST
PFgt'@c t€led
IIERV
E6do8.'fu Ellwi!.I .koc@
on Ccn
tic
Ac4towMnmts
My ri@F tb.d5 b dy suFwi@ Dr. Kh.lid M Kh.r,
yie Chr@llor PMAS Arid
Aeri.ult@ Uaiv6iiy, R sdpirdi &r ht rlt rift guidda, @Btalt slppon aod
acouag.n nt.'nF Bp.cial suppon of Dr. Am Khanuru Dad S.icMs,.e Bd be
o My Co+uperyi&r D!, Atikr Mlrwi and Dr K€bl.!h& Mszhd for i,leir
supFn ud guidae duirg.ll .ro.s Dy triq $pecially duiog ph.D. ThDl5 e
dr lo Dr Yeh Mlsdq 3 Elt for na h.lp- I atso wt to rhlrl D. tuhel
Q@d.!d DrQaib Ayub fortb.irE !!icd h.b.!d Fliliv. criticila in ddirg Dy
forgott
Vert sFci.l lblrks b Dr Alt f Hulei! witiosr whoE ny theb muld hrrc rcq
b€.n po$ibL. I woltd a&o lite to rh.* my @lleques Amb@r\ LubB, Sooda
Ati., Na4 [email protected] sb dd I!o&.1 Sb for their tiid co{p.ndotr and besi
vish... Th.d! e d$ dE ro Rdbdr bd N.fer bow N oG t[& &i.n ts ro oc_
sadi!,
o*
s itude ro &Dily Dy hotae Slolrt At i $r]id Bbti &d T.hi.
Bbri tu lh.n br &d s[port.ll rl@s Ey tif.. My sisi6 B!ji, Ruby.!d M@ tur
then suppqt .!d lM. TIErrs e d& to 6y hub.ld Sh.bbir, sio b alsys rtEG ro
help 8., ny !w.t daugltd NlmiE ud lovilg ea U6d thcir prtt! *!rc duys
I
hy dep
sp€ci.l
tlrnk
q4h.Dd
rb.!t!
to
d
dE to sll
rhe s[o h.lFd m€ in enhy duil8
lh6i! Eitiret espei.ly
do@B of i[e blood
AI-LAH for bi! @6trDr Rlho.r .!d btcsings_
spL'
th. riD. of oy
Ad v.ry sFcirl
ABSTRACT
Y{hrcbosbe of
idditrcdion of
1.86 Mb
the
w
w
lcFGrl uing e@oE. !€qtlEc dal! h€!.
3iogl. duhiplq @cdor .trd lh.i. dcgre of
baLs
liln
poltooryhis
Prtisrar Divditi.s of ildividu.l l@i nagcd
Pikistdi [email protected] wbes hlplotyF divaEity
rh.
ecidr tuh.ma
Mbl.d
polFo.Dtic [email protected] !. livc lai 9@
idbcs, cxhibitd v.ry low
B
t@
0-963.
div4iry
B
for th.
ilto
a
ilvdtie.cd in 755
0.2209 lo 0.7938
'It
-in
Moh@r, who dc
for rh.se nv. l@i (0.650)
ed
ore hplotype @YS648-DYS649-DYS650-DY565I-DY5652 haploryrE 9-8n7.ll24)
&@ut d for
brd lh. low
liG
vith
57% (38/66)
l4b
of
dildity
ofl[k
popuhtio!.
0.64, Th.
@. $r of ni@s!,r.[ir.
ooiliion
Itc
other
6t M
topddior KabEiri dlo
clrcda&d
E rk r, r*o ndrrionr @ i!
D@l@tile
EF3t tavilg g.@lb div.Eity of 0.79 ud s@nd in
nEl@dde
r.Frt
hrying g.Ftic dileniry of 0.E51
TIF ndation 6t€
6
m
@ foud
calcultLd to b. 3.l8xlo r aii.r 90
dd.tio6,
3,8%
AAb .nd
P{dsttri Dopurltiol
DYS650,
DYS652,
. p6L
gtrnrioN i.0.3%,
i!
which
foENic cdc work
on Y chroeosobc for infertility Esuis
5.E% AZF.
r &!r
itr Patj$ad popubrion
b. usd 6 corectior tutor whilc Bing this €l of meka
6ilyd!. D.l.lion napping
h gmwilg @U
i!
2% AZFa
d.lctio8 aspocibl€ for tule i.fcniliry
Ib. AZlc dcLtid q* foud ody
in
!
i!
gbup h.virg
I
CIIAPIER
INTRODUCTION
Y CSROMOSOME
lrx
i5
d.t min.d b hmtu by thc X ed Y<hrcn@n6. Fcmate of
rhe
htuM sp6i6 hlw tm siniL( X chron@me wh€e G th. hale hs. rm iqy
d'sinilr chtonoen.s,
egg
the
@i6 h.lfof $e
6r[d's sFm
in a
sy
$ar
.htMo$6..
is
d.,
Y
of a shod (Yp)
on€
in inc Y
dfi@t iod
t[& of th. momd]
do rot @r't in diloclMdri!
chenoffi,
ed . lois
humMs
8,hich is
(Yq)
a
m.
oru. S!.m
d.
f.rtitized
@lt! @ difteEnr
.rd n @t 6!ry rlE X o.
GpoNibt. for nd. qHifi. functioni
tt is .lways
paltu
rhc
y
@nsisrs
y inh.drcd, m.rning
rh.1 a
6py of hh own y chrchosoDc, which i! ulualty pssd
sau chog6 ell€d poltoorphism o@r
cnNhoM.
I.
holh€E DNA bd half of th. frthes, The gcn.ric contriburion of
fod€r paes on ro his $n
intaca Hou.rcr,
X dd on. Y (Mcctung. 19@).
orcr
.
long
r.riod otlinc
6 it is pats.d doh ti@ g6@rion b Sdqrion, fte
cheg6 h.lp in our undeFtanding oflh. huM dolltionary hisro.y (Unddni r, al,
2000) and
dc us.tul for fo@ic
Smiih, 2003).
analysh a! well as pa&dny
Cheg.s thll @umulatc
in nh. Y
t
stihS (Jobting
chmnoson. inctude:
sd
Ll,l lenioc od.Ltloa
Y cnDEo$n.
etul
in
of
insqtion, whid is Flricul&ly
300
b.* FiE
rh.t i3
r.nicul& Egion of th. DNA. Hmmd (1995) luglsts rh!1 dE
l!.,4ls clchat @t@d
. miqu. c@t
in
food lo b. YA!
$l[m AiiM
hln8
.tu
hilto.t dout
th. divds.ne of
popul.ric
ul
nosr ot d€
N sh.
dricn YA? b Gsinar.d to b. .
&id
wdd poplldion (tl|mh6od Hodi,
instion
6
!!cl6dd. poltro4h&@ €NPr)
SN?!
@
I
lbdq c
chon@n
is
in$ts
.xet .l.p&*
iNnbN
in
hmd
$,ho
DNA of
th. b$tu of in diiihlti@ in dp
1995).
SirSL
sit e *hcF
+vc Y
popul.rios
hdf . nillio!,alt,
@ft
hm$ ed chinpo4.!
yds.go, Chimps4
18E,000
jc, *iilc hignd h{!.ncy ofYAP
lFk YAP .ld.nt. Th@
l,ll
$
Fplldion 3nrdi6..,li! ir a 3.qu@ of.pprcxitur.lt
i4rt d irto .
in*nior
,i& polrnolPnbm (YAP) ir
pmicule i@lani& is cnsg.d. Sr.blc SNP'S @
El.tiEly l'& !.d e itrtlquat t[.r (ry @ t mcd G 'uhu. G6r polynorphisor'
(UEP', Bit.ry Flynorylrtmt SNPs
d
th. Y chMosdc pledc th.
86dic l.g&r ofou speiB (.lobliig $d Snith,
2003).
p.rh.l
.
Undedill
.r al
(1997,2000)
lsd
chmmalo8raphy 10 idendry biall.lic aites th6t
d.Bluring hiSI Perfommce
liquid
fom.d pasimoniou! g.nealogy ofl16
haplotlpes. Since Y chomosom. @udulltc rlbFqu.nt bisll.lic mulationd cne8ps
{irhour etuing p6r .v.nts, it is now po$ibl. to.oisttuct th€ h.plotpe by pleing
rh€m in dder
ed @Ntrud haploryp.s
for rh. poprlation. Distiibutio of the Y
chrcnoema div.uity bse<l on tn. binary
inporr.ni
hum pr.htroric
l IJ Mi.Mt
F!r*.B ha bm
llft. loltlorphbn!
Epat d ! v&iaue nudbcr of
in
! p.rticuld vdidl
titu
(or
of
2{ bc.
or
in fornsic
delle
(Moron
@ &d}!b {d
plir $.r @ $ndemly
tl6!8nou h. gcmmq TIE sDeilic nmb.r of
.ll.l.) 6ully minr !nchr8!d ftm ga.rdion
g..drion howcEr chDg6 do oeu. eh.1im.s &d lhc
irc6e
d.
nov.m.nB (Und.rhill ., ai,, 2000)-
[email protected] @ sinpl. s.qF.@ np6ts
EFlts
cmployed 1o qplain
ed
WiUs, 1999). Y
numb€r of
sd r
tlpau my
micD$Lllils a. Gpeially etul
dolurioMry rtudi*. For patmny tcsting
iivolving a dee.s.d all.g.d f.lh.r
ii
6p€
nalc child, E!noty!. of a fatnq
inf.Gd tom hk livins patdnal rlsriv. (Kay$ md S.jstila,2001).
dltabe codaiis moc th4 hund.Fd difl:cnl
YSTRS
ed
therc
4. n.dy
6es
m
population (Benson, 1999).
6n b.
be
Genom.
mor€ still
to be discoveed (Kaysc..rol.,2004). Thc pNgdm T..ded R.pc6t find.rh.lps in
&arching micrcsalellire ndk.!s, *hioh
to
tle
us.d fo! ils polynoahic natua in a
Micrcst llitc mutation
dd
aocnsic dalysis
m.d.
10
dalins th. branch.s of the
av.lut th. mMlional !n6
sbow thar
is
6t6 @ v.!y inpod.ll
a
singl. st
p
n@ @mo., thn
ts oiis r.t€r
of th.
r€eardihg iheir
tre, A nunber of
ni{olarlllit
rc|. in rh.
aitempts hav.
bcn
mlyss
hav.
s. P.di8E.
mLtaljon
@ltin8 ii itrctEe or dclEe of siDgl. EF.t
un
two sLp
nutnio. @lting in in@ or ddw of r
e
a
(web6 dd Wong,
l9l). vsjou
nodds n
w b.a
popGcd
*plaining th€ nutation m{htuism bs.d on thc polFe@ sliDpa& duiing
cplication. These nod.ls
a.
mostly
ba.d on Y niccat€llit s &d small sized
f.th.tson pair studies or ddiv€d rlom the lutosonal sTR3 (K6yer
Tr&llod.d
lymph@,4.s provide
sDl€
dl.,2AAJI.
fo.lab.Elory s:rudi6, which a. onsledy
ltoM in tisuc cul$E hbontori6 to nli lin
the @nsknt epply of DNA,
T@fomldon ed go}{h inrclvs tlE nhb.r of divisioG ehid
if, th. g.nstic plttem- Th.EfoE it is
*
impdlot
ro
* *nd
Mt 6lt
cheBc
stent dE g.naic panan
obewcd in [email protected] lin6 EpF*nll th. original DNA smpL orth. donoi. Ii poimr
towdds lhe inportance ofth. mut lion 6t. c.lculadon in tdsfomed cell linos.
1.1.4 MlilnGllir6 Polt oorphi3me
MilisEllit6
oncn
*tat
to
e
nomaUy 10-60 beF pri6
$vcal d@n, Chdg.s dsing th. @pyi.8
ftc{ucnily in niohareln€s
B&..al.
ldg ed
(2000) the
ihd
Y{peinc
in
nicssr.llitls
|@us
(Joblins
lhe nunhd of ap.lrs
pres t L
pl@ moE
./ al, 199). Ac@ding !o
MSYI .nd MSYz aE theonly knoM haploid
mini$t lliEs,
tst
dispbr
{ c imly hid &8re of lEucoDl di@ity,
nini$t llil. veisr cpat polyn @.h.in E!.lion (MVR-rcR).
MSY ir ln. otrly
Y<hrchoenal syn.m qna ln. ch.st ri$ie of hrg. nudbas of hurdions
nudicd
bca!* of
irs 6pid nudion
dr.. Mini$r.llite!
lowc.ful lool for O. inv6tis.rion of hutit@s
in
,l$
p@id.
. hlploid sy!r.n, .nd
.
for
6
b.
uniq@l,
lh. d.ring
ofp.t6r.l limrsB (lrlD &rl Prg., | 97).
ROLE OF Y CEROMOSOME IN POPULATION GENETICS
Th.
unchs8ed
3Fific nmb6 of EFrl3
ton
r!n.
.tploit d i! r'&itrg th.
ion to
r
in
&*6tjd
pdnicul&
hicrls.LlliE uully t@ini
(MoM ed willr. 199),
populdion hinorie
tqn
d
a
This
poFny i.
popul.tion 8..cti8. Popllttioi
g.n.ti6 lllv. shom th. odgii sd higr.lion plrr.n of vaious populdioN
@nsEGtid of thc phyloS.ncic
sp@ilic
phylosoctic
na
dlos th. inv.srig.rid ol tn mb
N .x!.$i*lt b.ing B.d for th. @nststior of
Thc n@bq of
rh. lonsd lh.
diflcnt
which
dpels of thc topul ion'3 hisroti6. Th. dl.lic sLcs of YSN?} .t6g widl
thc bpidly cvolving YSTR'3
UEP,
lrG,
by
wioG
d
impon&t in dlting
rtlcs of dift @t YSTR! .ls h.lp
linasls (Zdj.l a al,
.ll.16 show
tin rire th. UEP @s. d!. set
YSTR5 allcl.s. As YSTR5 hlvc
sNPs thcy
YSTR
1997).
dnc for Fniculu
r will b.
v.ry hish mltltio tnc
$. n.l. linag6 id.ntif.d
in
thc
tE imb€ of
cdp4d
d'.
ro
tt
SNPS.
All.lic
th. @nnruqio of h.!lotyp6, 3howinS
wi@t
by
oaPtkbtr
1.2.1 llinory
Pal<istlr cosists
0.{.w.F.P), Pujab
&d
., al.
origii3 (Mchdi
Popul.ilon
of fou! povine,
sindh that
1999).
ft6.
4
hdo-E@p€n, but th.y
larglag., BEhui ed
Pakisid
SincTibet
a
ha m
@ b.scd on linglisti.,
>60 l&gu.g6
rle
estim.ted
hhu
.!lt!nl ed librl
(Gtin6, 1992). Mon
ircludc m
r l&gug.,
ollhe
ieh!., Bchastii t Ddvidid
Bdri.
of oed
population
169 niUion
in
2007
c
(WikiD.di.). 'nF Bal@hs @ the main populition of adochistan. Thc B.l@h
@r4nt.l.d
t'&.
in thc
Kala! sibi
th.m b&k to Sy.i. vic
@nsid.Fd to b.
Tl.
ed Leli
!n. ft.
bdeE@pd i.
disrricrs
ofQ!.na Divisim. Thcir u.dnions
hrgu.g. spok6 by theh
is Balo.hi,
s,t'i.h is
sd
ris
origin (cdmcs, 1992).
DmvidiM cultuE blossoE.d ovcr lhe ooluriA in Sindh
th. Indus Vall.y Civiliation eund 3000 BC, *tich
oftrib.s fDm F.srqn Eurcpe. Sindh
ChFd Asi&
dpiG
Esulline in a distincr
populations .onsist
sucb
dd
s
.
mey €thnic srcups of div@
inhabitcd by
Seups
.ffiliaios (rie l-l) *,to Adk
L.g!.86
@
Bd@histan, West Fontier Povin
lh.
E
wt
ofid th.linal
P6is,
gavc
inlo de.line by $c
sr@ tor Middl.
to
eivrl
E!$d
md
CEks, Ad!6, Turk &d Af8hm
unique cultua .vcn b.forc
ih. eival ol IslM.
Sindhi
of Mohdnls, M.os, PaNk dd indigflous Sindhi populltion.
Moh.nn. rdbe is spE d over Makan coasr in Sindh
dd $uilem
Punjab, Th.y
4
,le
idcntifi.d
s
rhc M,llal'or
lish.db.
p@vin6 th.k chn o.e&ianions
hwe @nn6tioN wilh Hindu
-Ite
M@s havc thcif oiigin
Thc o.igi.s of (he
prcph.l
Zooat
lhe
th eirlry
rhe
"P6|
r,
ell
6
Hindu
cat6
dalmatcd
ft follol6
lhey
c
Iruir
Sasie.npiE
* mpl.d
th. p@nt popuLrio
not n
v.
sll
d@u@re<t hisrory blt
in
w.e c.lled
(N@vuuy
[email protected] b
@ rh. desndels of lh. Dqd! fom cc.F.l Asia (Biddulph, !9?7), dd
4
Pr€-Aryan Crcut (Wikip.dia, 2007).
rnori.s for tn origin of this popu'alioi, $.y mid't
shc clain
.,
be d'e
to be dF de$cndena of Alcmder
200?). In som. srudi.s no
genetc
of th.
(m.&ing "Aom lrs"). Ev.nru.lly they hded ro Mlnbai in lndia &d
-ft. Plthu @ th. mrldt lag6r (p.triaEhal) ethric
(Mssoor
io
and not only from ihc Rajputs
A.D. Th€y eltled in 900 A.D in cujadr, India, whde tn.y
consid.cd lhem
Isdel and
Sindh
Bml6ms, M@r, ht5, &d BlEti'6.
wio migal€d io Indi!.ftq th. collapp oftbe
197). B@sho poplLrion do6
some
such
ton nuy
K@chi in P.idsr6, Iioa wn
some lhey
od
llsblc $@ of th. Rijpu|l, but olhcc sm
6tcs
P6is de
M@3 mostly inhaln tnc Punj.b
gd.tic
evidencc
srcup- Th.a
e a mt
d*cndcnts of rhcjc$ ol
hinelf (Khyb.N.lch.on,
h.v. ben found to ftUy
suppon this th@ry
di., 2004), whil€ the sludt of th. Fna$ e, al. (200?) haw foud sone
sidilaity b.twen Gckr od P|ths. Kal6h re m ahnic 8ro!p of
the
Hindr Kuth, Ftlung in the Norlh-W.st FEnli€r Povioe dd sp..k r h.mbd ofth.
Drdi. finily
d.*.ndetr of
direct
ALxmd.r
lhe
lmguag..
klsh
hiNlf
Son $hoh6 hlw speuld€d
either G@k
sltl.6
$c Kal6h
of the AlexMd.r th. c€at
(hnp://kalahp€ople.c@)-
population
rhal
Howd, l.@t
s
b.lhe
might
lmy
or ev..
86etic lening
mog
ha shom th.l rh.y e, in f&t, a disti.cl (sd FlhaF
lbongi.al) population and fallclo* to
t!.
Gst ofPakist&i populstioN (Firqal
€,4r,
2OJr.
Tib.i.n p@tle ud Dedic bib€s cffi. to Baltisr.r Fior lo civilistion, lhee
gow *ntullly
$nled dou, 4d oeig.d
B.lti p€opl., howevd,
e
nol
.ll
s
th. Blhi p6Ple. Thc dodh world
fro6 lic lib.lan stock. With the
mdy ods rlibes nmely, Ka3hdiris, Ar.bs, Pcreie od Tu.l5
CaduaU, ndecd with lhc locll popuhnd,
bM.
Baltis, Punjab Province shows
r
cd.
Ato s@nd d thid
passaSe
of time.
to Eallirr.n
g.n nlion lh€y
@mpl€x adrnixturc or c$1e5 Md groups
(lbbetso., 1883) dr@foc no 8lnetic popllalion study haE includ.d punjlbi
sludy
i. d.ir
s fs (Qher d .1., 2002).
@i.d
Stodics b@,
Pakhar
I
dd
by Qamd
(P),2 (A), 3
out lor
d. @nsdction of rhe hum&
ryolutionary lEe in
a al. (2002). Wnich hrv. shoM th. hilh frcqEncy of haplogroup
(Rlal), 9 (J) dd 28 (L) qhich loserbo make up !o 79% of dc total
Palist ni popul.tio.
(Qdd
€, al., 2002). sMies ba$d
esrinate d.r. h.v. show. rhat c.ntBl Asi.n popuktions
o. nicmsar.llit frlquacy
d
$c m61 divese
it
Elhnlc Gmop.
Ih
I.rr
rxE
Is
FEl.l r/gtk
l-Ad
ol
niqrdilb gffF
|! P.rid.!
l0
Eu@ia sgg.ghg llut Cotal A3ia paul*ioni
gcn.tic pdt
m age
wirh ecou.rs of
Indis subcontircdr (s.i.ls:tad ., dr,
tJ
$dnr
e
the
old.sr in lh. eSion dd th.
Indo'E!rcp€n
q.n6
@nqu.iih8lh.
1999).
ROLE OF Y CHROMOSOME IN MALE IT{TERTILITY
Almost
l0l5%
couDles
@ inf.rtilc (hilto on@ire
!
child) &d lpPFximt ly
((qtd dd Bokd. 1999). hfstilitv ii
nal.s may ncve dif.enr @s like .nviMn€ntal or g...tic Th. t quentlv
ndf of
l|)* c|g
msl. fa.tor is Espotuiblc
[email protected] o@ of mlc ilfqtility
inpalriog
st matog.n6is,
inrctving !
sri6
of
is
Y
chrmom.l d.letiois (M4eur,
2004) by
spdmatocEnclis n . @npld [email protected] of sp€m Poduction
lsious sLF it t[e sinaf@u! tubuls of th. t sljs
(hnp:/
rw.u.aleay.c/Uofc/.due.Vvinul6b.yc'ldcv) Nom.l humo nale
t3-90
)96
6
mok I billion spcm a dly ed 6du.tion in tt m Poduclio
A@oding to senen snalys
is
inf.rl ility is caLeoiiFd inio:
R.duc.d spqn poduclion NullinS in spm count <5 nillion spcm
elltnl
ll
Aaospemic condition develops illhqe is
th.
sFm!
!.rh howewr
Thc hul@ Y
80
pMss
thc
of
spm
poteis sohe of whicl
e
b.liwed tnlt lhe Y chMoeno w6
g.ne, the ex.detdmining
Sonada! devclopn€.r
in
n.Fb
or it
in
Aponsiblc for
2003),
gm
dd
b. du.10
sullio!
Ym
1990s. The
bd
pliB dd
c.ll detelopDar od
ago,
. $n.tic @Llmd .r4pr
fi. sly
*
production is nomd
g!n. SRY (lh. sxncoEmining
.honNnc), s,li.h ws idatifi.d
.honos<re
.Gq
cheno$m. mtains +pmxim.Lly 60 million
Gini.nec. Gp.m.log.n6i!) (St l6Bty .r zt,
ldds to
problen in th€ production of
in s@inif@u bbllas @sing spdbdoSaic
ob6truction in rheir
dcods
sy
n6nt biologists
for
o*
@gion
import&t
on fie Y
plts.@ ofSRY poleins
the ngl. pndotype sugS*ting thc rcle of Y
Gnility (Schl.g!|, 2002).
&d Zuii.di,
(1976)
w@ lh. firs. io htToth6ia . oRlrtion
bd*a
Y .hrcnosFal d€l.tions &d m.le inaeftility. Ctto8.n€tic r1udi6 in sl.lile
h.lpcd in rh. lo€liati@ of
.
men
8de @rmlling sPhatogpn6is, cf.Gd lo
6
u@spdmh fador (azF), qhhh h6 AzF., AzFb ed AzF. cgiore (vogt ,r dl.,
196), Th. cgio6 rlonS with sp&ific 966 b shos in Fig
our by
Bo{n ., dl
(1998)
ed FoElta d ai
(2000) Y
1.2. In tbe srudics
chonosomd s.n
s
di.d
lhat hav.
l2
I
I
4F
l:
?@
<#:
I
:g
I
I
d |bdneA.
"tddi!&
rfit.drZF. n{i&
Iirl'2 CrroSd;&-tt
lo.io
of .AE
.
oftb
Y
dt @t
ll
b.a lint d to dp@aloga*is, Miding in AZFa region
cxpKed DFFRY, DBY (DEAD
I
(Asp-Olu-Ala- Asp,4lis) box polypeplid€, Y
choooshe), ubiquirouly E6erib.d 1.tdrli@Fptide
(UTY)
Dolil
nd
ubhuitin spdifc
ms
(RBIIO @nsis$ oflbout 30 @pi.s oD bcnh
inplided
in
sd
s four
ga. Gnily (M! .r at,
$e t nis. Compl.t ftap ofAZFb
dt ib
gcnc, Y chonoson.
(USP9Y). P-r{A'binding
ofrbe Y.hronosnc. USP9Y
bse psn d.l.tion in this 8en hs
daL infdtilitr (Sm./,t., 199).
AZFb Esion is RBMY
eFlt
Fo{ss 9 d Y chrco@dc
is a functio.dl esrc in tbe AZFa Egion
bd
includ. ubiquilously
1993),
The
nlin ddidrn eE€
wn@ .xpe$ion
ovd 3.2-Mb,
ad
is
in tn
qi.ided io
ovcr 12./. of ih€ rsion
@NiNof rcped.d6.qu.nci0*lin.r,1,,2001)
TIE go.s
Pag.,
199}
i. tE
AZFG
i.
o.stis
(lrln
!d
Dcl.ted in a@spemi. (DAZ), BPYI, BPY2, chonodonain Fotein, Y
ch@nosofte (CDY), PRY,
ed x Kell
cgim haw spci{ic expBion
Lstilsp€inc bansiipl, Y-linked I Md 2 OrYl, TTYD
bl@d 8rcup pE@Grclatc4
copi6 in thc AzFc cgion. The tuin
Ylirr.d GXRY), all .xisr
ddidat. g!r. i! lie
cluit 4 nuldpl. @pi* of DAz (>99% id.ntic.l
in DNA
in multipl.
AZFo Egio, is lh. DAZ
s.qu.ne) clusld
have
ben
AZFc regions de dponsiblc for 8 %
cB
of h.le infenilir, rcprcenting the non fr.quotly del€t d Egion in infcftil€
na
Eporr€d
i. AZlc Egion.
Del€tiohs in
it.
Y cnmmemal DAZ
gft hs a tu cdoBl
homologw DAZH
d ec6tr.l Nto$nsl g!r.
chomosme L Analysis of th. botn gen.s indiooled ftat
naw
gim i* lo the Y chon@nd
o. hutu
DAZ e.n. durinS mlution sugr6ling thc
impon nt evolulioney pr@ss ro prcseNc th. f.niliiy
g6.s (Sdoia 4 ar,
1996).
Anoih.r inpod$t €volution ry adaflarion h thc pc.n@ of multipl. copi6 of th. Y
g!n.s,
s
coti.s of
$e Y choho$m. h6 .o duplicak hut ils p6liodrom.6 povid. b&kup
irs
critidl gen6,
so it
ce b. apaitld
g.n. h6 foui @pi6 dd d.lcrion
Fe
in.r,r
G @ist
ofoy
wh.n dm€ged by frdadons. DAZ
oo. or rwo
apy
is
€lled
a
panial dclctio!.
€005) suSgesld lhgl ohly p.nial MFc d.l.tions ENrtngDAzl/DAZz
d wirh sp.morog.nic
impsifrot rvbels thoF eholiog
DAZSIDAZI
m.y have little or no .fTed on fcnilily.
Haplogoup ssocialion oflhe Y d.l.tions hrv. b€en docudcntd in various
rcgionr of tlE
wrtd s
Rcpping er
al
(2002) found
6al
lll 9/g
dcla.d chrchosn6
b.longed to haplogroup D2b, vhich is ftcqu.nt in Jspan. The grlgr
lhe dcldion
of.
l&g€ portion of AzFc cgion
dd ! k om
d.l.tio. involv*
risk f&tor for m.l.
f€nility, l1h6 ben susgBted that this erl8r dcl.l.d hsllotype mighl bc
subf.nility in
J.pe. Fensd6 a
d. lEen
fo!
al. (2003) found 1.6 Mb d.letion in lh. Y
chono$me of hrylogroup N, which is wid.ly didributcd in Eropc md Asir md
tn
€
is no
indiolio! ofr.duc.d f.rtiliry
th. Y chonoson.l d€l€tions
10
in this cgion. Sci.ntisis
!d rying
particuld hlplog.ouP which nay
indi*c
lo
coii.lak
such gbuPs
l5
e
al .ist lo d.velop infertility. nEefoG th.c is ne.d to figue our the Y
chrono,orul 5utus of
Patjsran
'nfenil.
rols,n
is inhalit d by
Haplosmup studi.s have ben
18
tried
Palisuni popdlarion!
dfffil
.r di.,
1999).
al. (2002) to
asigi
ednic ercups (Mchdi
out in P.kisbn by
Qder.t
them vaious haplogroups howevq no study is dvlilabl€ ro show
lh.*
haplogFup! with infsiility. Th.E
b.tsq
hsle infcnjliry with
Our
ain
is
als
pari€nts, by napping
to
wlos
a n..d to cdry
o
a@idtion studi6
hatlogoups.
fud out ile pcMtlg€ of AZI deletim6 in P.Lilt ni
d.l.lim
th.
i.f.rtil.
in AZF!, b End c Fgions, therefoE lhe stldy ha h€en
ldenlify polynoAhic Y nakqs,
Esrinltion of
wa
th. .sciation of
mut tion
dd
d6
to use theo to d.t mine th€ poPulalion
in gtowins
ell lin6
Pr.pae 6 calaloCue of nom6l snd pathogenic
cnomosonc in inLdiL Paldsrini m.lcs
dd
also
pdracule h{plogrenp. Thh wiu lad
Bine Y sp.cific
CEnetic variations
of
1be
Y
lh. deci.tion $is deletion
b rh. d.wloth. ofgtneti.
wilh
mt
r€st5
ior such p.li.nts md povidc scl.clion crileria 60r inf.nility clinics
involved in ss islcd Eprod uctive precd uFg
t6
R.EVIEW OT LITERATUR.E
!.T IRO}XRTIES
The
difi*n@
OF TIIE Y CSROMOSOME
b.t*.i
th. t*o sx.s is tt€ pEenc. of Y
hunM mle (Mcclun& 1902).
turction.
n. qpBioi
chrdosm. i. rh.
of mlc [email protected] is it! tuior
HlnD Y chomosnc cohpds of 60 nillion b& pri6
(Jobling
ed
TyL..snid\ 2003) .nd @aiB .bou 75 p.ot in<oditrg g!!6. The e.G
@U.<tiv.ly dcod. r,r
lad
27
kroq
didin
l
Fot inr (Fig 1.2).
0997) d.qib.d 6e fircliodl speialidion of
rhe gd6
t
h. dd P.g.
in lhc
p|ffi
of
spamog.n6is. Y chM@m. @sisl! of two p.rls, $c psudo{urGomal cgio!
sti.h .x.ha$s
boM a nal.
or
m.iqis, bul
etion.
MSY
failue.
The
scgncnts duing syDs,sls
*ith
rhc X
sp@ific rgion (MSY). MSY d@s .ol
homologous Fombinatio.
b*n
snown 10
chomo$nc &d dE othd
@Dbin. wilh G
Frt
honologqc
t*.s tle. bawd uplions
of MSY
8.n B& d.laion!, cohmonly @ulling i. spmstog.nic
nah 3p.cific Fsion caiii6
!
fcw s.n s
ed
lhc 16l of h .ompds.s of
nicrcsatelh6, minielcllit.s md Aluel.ncors (Schl.8d, 2002).
l7
2,1.1 Y SBinc Micosrt llit
Thc Y spccific hicrdsllclli&s or shon
bs
pair unirs
ed
th€
l!.d.b
Ep€ats (STPi),
nub.r of rcFd uits vri6 ftom individud to individud
e
shoqing 1hlt the nicresar.lliLs
porynoryhic (Lin
dd
Luty, 1989; T&E, l9E9).
Amdg hicos&llitcs, rhe dituclolid. apots w.E rh.
Mi.sfield
.r ,l in
@sisl of2-6
1931.
Micclt Uir6
e
fr{
to b. ftL.tified by
ini.6pe6ed rhoughout
lh.
non-
ps.udoaubsonal Fgion of lh. Y chlohoemc (Rocwq er al., 1992). The nunb€r of
ep..ts .cmliN tlF
same durins
dF sn. This pop€rty
S.jstil.,
@nciling
2001)
th. tmsFislion oflhe Y chomo$m. fron frthq lo
[email protected]
ed fomic
$. gd.ric,
is cxgloii.d in
linguisric
dd dh@logidl hinori6
ud Hdne,
(2002)
(tyleFsmio
deribe(l th.
N
of 9-l
reliable
populltion siz. Eay sh&
a
num41h, Fquic
of turuE
1996). Y chonoem.l !€sqrch
1999r JoblinS
sd
I micrcs&llh.
Mk
!
a
iD
$ud.
fot
Katsr.,
a?,
provid€d a unique
6 fo. fomsic u.lysis,
but
foi
l&gc nudbcr ot mi@!.r.llircs
infftne, vaiols studi6
@mmoi h.ploiypc
|!ssh
hs h.lpd
Md Tyl.Fsmith, 2000).
.volutim&y srudi* dd popularim ga.tic ltuda*
@ rc.d.d ro ddw
of
ed idplenotalion
t@ing ll1. evolutiom.y origin of th. huns bcing
gcnctic did.nce
rd
arlysis. S.iatbrs har l@gni*d ll)c ncd of
linSlislt .rd Ba.ticists to int @t ir tlE d.lign
st'ar.8i4 (Mitch.ll
Fldniry tBring (roys.r
harc shoq rh.t a l&e.
Zcrjal er al. (2003) !.pon d thll 4yo
ofpopulaiions acoss Asia shee a omnon h.plotype for 15 loci and in othef $udy
IE
14%
of the P6i popullrion in P.kisla h.vc b@n shoM to shG a @nnm
h@lotlTe for 16
2.1.2
nimt llic l@i (Mohtlddi.
Y SDeific
a ar,
200 | ).
Miliei.llitB
e
bms of ihe variable
nature they aE used in popularion studi.s, Jobling €r ,1, (1999) M rhe {irn b disv.r
mini*t llit MSYI, @Ni$s of e u.y of 50,115 cpeds of 25_bp units h&ing
Minisarellites
v&iabl. s.qumcB due ro
very slow dolving Darlc6 and
b4
!@nd ninisd€llit (MSY2)
subnirution. Blo rr
a slow
al
in the r€ar 2000
disova.d th.
dolving m.!t6, @ntrining 3-4 copi6 of .99-
Alu insrtion is al$ impordt polymorphism for populslion study. DYS287 is
3
Alu elemenl lepofied on the long Alm
t94).
son.
The YAP in$nion is a uniqu.
188,000
yes
ago alt r thc
I95). tlk gDdy B bed
Euopan dd
ollh. Y chromoede
FIam* (H4mq,
.v.nt in hune hhtory, which h6 @urEd
div.i$!.c ol huM ed
on thc dna
by
obtiin.d
[email protected]. Glammer
frm AfriM, Asraue,
cl'imprc smpl6. Chimtoz. sudd N
YAP
i€
tapan@,
od moc
of
lh.
l9
Ari& popll.tions .xepl
YAP
+t
thc Japd.s.
b round in sub.Sai@
l*k [email protected]. wllil.
hidcar ft.qu€tur of
Atiqs ,orul.rion! (Hmmo dd Honi.
1995).
2.r3 Si.gbNu.l6lid. P.lyEorDlird.
bin.r,
Poinr
muorioB thlr
turt
d,
e!.
*nici inclu& be
8rd Elropoen iMnioN.
htplotyFs, vhich shows
l9a;
S..ios
.,
aa.,
m.rt6 h.v. rm
(1996)
lmy
ec6l6l d.F
rcrld.
fo( Fovines,
on rh.
on rhc Y
1998;
c.tt.d
at. t997
.[cli. ![r.s
GhrM
of bi.llclic
sel.6 dd snjdr,
a
linete mat.
chDm$mc hs b4n rtudi.d
on. of Ih. no5t arhricdly
S.i.lgrld ./ at, | 999). tatisrd hrrc
rontici Provine (NWFP), B.lochisrM
F.d.Elly Admiiist dd ionl dd Tdbll AEs (FATA).
ed l&8ul8.
(Mchdi .r
dl,
s
choi@n. s uniquc.6rsr
qd Prtjsta, F @nr
eidinS in Prl(is&n bclong to rppbxiftarcly
cultuE
thlr pdticulr gtuup
wirh Espet to rhc Y
Indir
e
us.d ro @Bt Da
rh.l rhat d I ddi!.d sll.t. desnd liom
thc Punjab, NoRh Wcn
6 rcll a
Th.y
by Und.rnitt .r
(0) or ddivcd s!.r. (t).
ed g.n.riqlly dirc& cgiotu (n ijuftt r,
Sindh
hislory of
H.ploqF @.ruct d
dif.dnti.rioi
agions of rh.
Mt'
Ths birry/ birlLlic oskc6 G
chmomB
Popul.tion
s deribcd
subnitltion
tll. lindg. r.d
2000).
s6r.5:
.hon@n. @ re
de.ib.d tn. bitrr.y lolynoDhis
implying thlt all
in
o. Y
18
!d
P@plc
diff.cnt.thnic 8rcup! bas.d
1999). v.rious studics h.v.
bs
etri.d
on
out ro
20
r@
th. 8.naic origin or
ttut Ih. sulhw.st
o
vrio$
Plkjstari ahnic 8osps.
popul.tiod of Pdi.r.n @ry
!
Afii6
ep6lc
Aftie hiSition oarh. P*isi.tri fopul.iio.-
out of
t[@ Flllrtionr md crhro-hisloric
Nofthcd .thnic g@ups includc rh. Parh.lr Pujlbi,
K.lsh ed Balti. [email protected] .rhic soups
od
Sindhi. Noth.m
P!ki!t{i
a dr
(2003) u5.d
l{9.
tu hmo cElutiodrt !8.
e
Hrzt!.
mc lnd
nunb.r ofrh.
Thcy found
099)
3hoqld
d.ta $ppotu
Btosho,
t
Kqhniri,
rh. 8.1@h, Bdlui, M.la.ni, Mohen.
.thnic gDups (Prthu, BuNho
ih. [email protected] of tnc Gdls, who
Ayub
aa
YAP' Y chonosom., wnich
i.dic!t6 dc
origin of
Q.ro .t
th.
od K!bh) cl.ih
Al.x4dcis invsion
in rhis E8ion.
&iosd l hicoln UiE.lrr
rh
cloer to Euopa popul.rim. M.!Mr
io b.
ro @n5rtlot
non of rh. Pa*isrei Fpuhrios
G
., al, (2004) us.d Alu in$ni@
polFoQnisn ud auhidlclic mi.rs.r.lliE d!l! ro inf.r rh. gd.ri. Fl.doNhip of
tfis
potulltionr witi
Popuhtion G
YSTR
ampe!
bst c
CGkr.
to
Burusho
hd Pnne
*E
d. K.lah. Rstly Fier
found
1o
rh.
Gck
.r a/. (2007) us.d dE s.t of
ro tunhd n@w down th. g.n.dc disre@
Two Dopulrtiois hNing @mnon hplot
cloF
16
h.tq6 Pdh{ .!d
t. n qu.nci6 d
rhouShr !o b€
cloe
&d h.ving simil.r bioloSiql hisrory. Hinori6 of vrrioc ,orld pop{hrions brw
b.cn
Belcd
by Y chromGotul b.lysis, showing rh. g.n.tic cla,tionship in .v.n
gagBphislly distinct populalioF (Jobling dd Ttl.rsmith. 2003). PHYLIP dd
2l
.e li. @np$er pogMs that @ ued for th. conitructio. of
phylog.naic ra ba*d m ditr dl Y fricoscuic tequencics dd popul.lion
CENDIST
grclPs.
Td6 als
based upon
dereloped
th.
i. lh. cal.ulaiioi tlc divcEqce tine for $o
help
dbtance
be$en tn. brrn€s. Nonenclatue sysLns have
b@n
to 6sien lhe nm.s !o wiols h.plogroups by thc Y Chrcmosn
Thc YCC
h
a Crcup oa $imthrs involv.n in @llabo.arive cEon to $udy
gcneric vori.tion on the Y
lins deliwd frod
chonosoh..lts aiEs ee
ro eslqblisn a lymphoblstoid ccll
individuals Eps.ntinS indigenous populErions all ov.r lh. world,
md lo povid. DNA iel.re<t tom Ih€*
ell lin6
to invgrig$e polymoQhis on rhe
Y chomoeh., Thc YCC .lso.ins to 6rablish a @mmon d.t
Esult! ol typin8 DNA5 fion
many
populalioB
rh* all
be
@ni.inin8 tn.
lins. YCC lins ro includc $c rcsulB of
lei a po$ibl. (Iobling and Tyl.Fshith,
6
1995).
MICROSATELLITE MUTATTON RTIE
Micosai.llir. loci
all.l€s, bd
$.
have
eldiv.ly
hiSh nutation Etes
od lag. nunb.6
of
larg.sl alleles eenemllt hav. ferer thM 100 repets (taulz, l99l).
Polym.rde lemplaL slippage muldtions hay occur to chaner de nunb.r ol Epeat
u.its in a micoeellite
Micros.llit
vari.bility
lss
is
(schloltear
not onlt a
ed TauE
1992; viSu.ra
endion oflhc nicroerellnc
.r
al., 2001).
motdion rat , but
22
.ls
populalion hido.y
2002a).
Undqhill
€r
single evolutionary
.slutionory
zl
isrmpf dd Goldsr.in
dsib.d
(2001)
2O0l )
rhlt mo<Lh
@ldi2nio, ed difr€En6
wilh
in
to nubtions dudng .volution.
vdoc
swivll
(
$ogEphic
1997)
of ftultigeo.ntio.al pedigrces, covqing 213
n
ar
Bdt
of nultipl.
ag..xFrsio.s"
miSdions.
is the
of th. populltions. MimsateUit s
H.yq a ai
er
hmd spci6 originlt d frod .
dent Di4city don8 hlmu popul.rioN
eqL' Miar.d
illd
&d s.l6tion pNu@
e.lyzed
the nine loci in
@ subjal
d€sndell
iosis and found four nutrlions
n
4
.vq.ge mutation nle.slimat of2,lxloJ (L.i&d Sun,2003). Boyer et al- (1998t
leponcd rhe sponlar€us
nuratiors
nltlrion
ar
Esuhs
i.
6 3.lxloi4.8 x lor
x lor to
1.5
x
IO
I
sain or loss of l-3
tne 10,000
huhd
livc ni.os.lcllit. l@i ed four ninisateuit lmi. Th.y
elcul.r.d nuEtion 6t6 G i{ring
O-5
ofdinucl.otid. rcpqls
/ c.ll/ g.n@tion. Sajdtil! 4 cr, in 1999 ealyz.d
gmlin. r&sdision .!dts
nom
rarc
iw
!t ninisst
0.2
llit
r loj
to <
3.1r IOi d nic@rellic4d
lociEspcclisly. M.jeity ofin. nulatioN
Ep.!t uniIl whcr.a 8ri! ot
loss
ofnoE rhe f.w EFrr
2,2,1 Microsalel|n€ Mutatioo Mod.lr
Thde h6 ben much incFn in obtlihing
svolutionary pro.Ess
Krugltal
er,r
.votulionary
d
inpoved u.de$mding of th.
thd Foducc micost.llit v&iarions (Jorla
1998,2000).
Mo@\i.
1975
dcwlopcd oosr
oftn
a ,r,
197;
nod.ls ftr th.
pro6s6 thd podued tllclic nlqu.ncy dintibdo.s Tn.e
models
m
nosily bd.d o. &alysis ol 6dom-*.lk modcb md
sLpsi*-mui*ion rodcls
in "st€ps"
bcde.lLlq e sudcd
Nltin3 in inc@
r
tn
k oM @ll*tively
nutnc upw.rd
dccls. of micoslr.llne
''lin6-wirh€n tanf' ftod.l. F! ed
on
to
e
ChakFborry (1998),
olhd had desfibcd that mutalions do not occu
dd
i.
leneth
hger
6e
poes the prcbl.m $al $me populatioro
those
e
il ws
2000)
lropoed
{
calen
il99t
the oriSiml
expeted to
lwh
tudom
auele siu s
,
gNup of
$8g*Ld rhlr poiot muratioN cm @cu.
.vollnioray
pe*
i.
$
st ps md suegesled.
micosat llit6 b@khg tn m inro sh.lld unib (B.U iod .turh 1997).
(198,
ed
obedcd in n.tuE. A hechanin wa popoed by
sidrins in which
dMwd
or
Moftis 4 a/.,
ddom walk nod.l for nutltion6. Thc non.cquilibdum mtuE of
wlk bod.ls
a
wirhio
Kogllit
€t aa
toirt nutlrioN Md slippag.
wnich
!.r simunlMNlt knoM s r M4tov ch.jtr trFd.l.
IGy$r
er
at
(2000) @nclud.d thlr multion
ed Y chnmmnd micrcsr.llit
msheisd is see in
both ryp.s
snalic @lls, bolh in
subs€qrent culturing
i5 @nsin.nl
of micost
lymph@tt
nr. in .utorcm.l nicosatellites
&d
llit6,
thus th.
nulllion
Instability
ws
gen€aling
rlso fohd in
s diecrly cnd ihnonalization atul
ofihnorrali4d lymphocltB. Mcl'gar rr al
duing
(2000) r.poded the
sel€tiv€ pEssw on codin8 rep..rs. TlEy comp.rcd th. edchm.nt of sp.cinc qpe of
nicrcsal€llites in codins Egions
b
b€
ed
in
th. non-coding Fgioa. Coding rcgios appd
mdtr mucn $rone.r sd norc sp.cinc @nsfiainB. Tri nucl€otid. &d Hem-
nwleoride ep€ds
e
found in
qB
in both codin8
ad
non{oding regions but olher
Ep.at typcs
speific
e
much
fFqsnt i.
*l6tior lslinn tm.
Sibly
.t 4l
non codinS
shiB
Egiod tho
in codiig cgions, indicatjng
muilrioN in coding Egions.
(2001) dcvcloped
r
new arpm.ch !o
M!*ov
chdin nod.ling of
niNstrcl|ne evolution $owh polyn.@ slipp.gc 5nd inlrodu.td a nd nod.l:
''@nsic.t-slippsg!-de'r nodel, in
*{
.b tha. is no dep.ndoc. ofslippag.
nicrcsat llite le.gth, Thc au8gestion thar slipFg.
ald
6t
o.
aE zerc or close lo erc lor
ii
v.ry shon [email protected] ( 14 Epels) ha impodo! impli€tiom
unile6t nding
{r.
n*hani.n of polyn@. slippls..
Diaingd ud S.hl6tt
os$
d
(2003)
.t
li.d tlE p.mur.tion-b4€d slrinic
ln. nicrcsareuitc disribution iD l0 conpl.icly equ.nc.d
with pFvious reporls, tb.y ohsned
a
ni8lly siErlifidl exc$
thrcudoul g€non6. B.!.d on @nput
r
rinularions
genomes. Consisi.nl
oa
d.t
b
lo.B
nicblat lliks
inroduced
a l.i8rh-
ind.pcnd.d slipFg. (in&l slipplS.) nularion mechdism op.nti.g ar Ep.!!'
Mictu$tcllilc alippa8 mutations
ce
pr66s* $at follow.itq stontaroB
bc
th! 6ult of hhtakca in DNA rcplit
or induced DNA
dM.8!.
extcnsion of s prima-Lnplab @DpLx @nilining a l@p
it
SliPPag. m.ds th.
.ith.r th. prin r
or
tehplat strdd by a DNA polyne@ (Kunkel &d Bebcn.k, 2000). Sponh.ous
nutation rares vary widely b.twed typ6 of Eicrcstel|n6
lci
wirh idcotic.l
Epet units.
Su€b
nltllions
have
dd
b€twe.n ihdividu.l
b€q rcpon€d not otly in lne
d emtic
gem lin6 of vaious sFcics bui al$
Oxidlrivc DNA dahag!
mo<Ll
sy3ts (Jekon., al,
ce
lead to micrcsct
1998).
Ci!3Eltc
mdarions ih cells wilh int.ct
llit. lliptsge nut tio.s
in
sok @ eus. hoy tyFs of DNA
drn.gq rh. obw.d r.n .nuclaride micosd.ln€ i$tability in lung 6s
Iinked ro this expGuc (sia
r,t,
f.cto6 phy. role in obrfrtd
bnStn
dd sleinc Fquae
fleking
2J
squo6,
ROLE
Y
Ticpolo
Of
Md
Y IN
ii.
1997).
Th* euls.leely
nihsclli|
of ln.
q.r!
'l'ai
iis
go.tic b&kgound
&d Zdfatdi
nas..y
(19?6)
for
nultiplc
cl$moMal lqtion ad .ssiatd
in which
$c mul.rions re m.a!rcd.
fts
h.lc s.xual d*elopnent 4d sp€maiog.n.sis.
suggErcd thc
ed mal. i!&rtility. ltcy fourd .leletions
posibl. olc of tn. huma Y
Funiq slysis of
of
ths
b. M Y{nrcn@he
lh. lsminal Yq rcgion in 0.5% of infcnil.
dcnovo deldio6 of dis6l Yq by hish-
@lrnion beding tehnique ns id..iific! ih. spematolpndis l@6 o. Yql1.23
1be
@spcimia factor AZF, Mohcula ealysis oipotients wiih nicro deletions
shoM th@
be
INII&Ilr.fry
chon@m€ b
(.4Fnic).
indi.rtE
hry
frurdion rar6, Thce facto$ ircludc tnc
chhdoem. in 3pmatolpreis dd d.scribcd r oftlation
mn
s.v.El
dif.Mt Yqll
difi.Enr phs6 of lne
suEEgi@
Fo6.
tn
6
has
l.d to sFmlto8ddis diNPtion
a1
AZF locus is divid.d into tn@ norlvdlapping sub
regions, AZF
{d
, AZFb
l@B b elive during
AZFC. E&b
8!ft clf d.velopn nr. (vo4,r
a
diilq€nt
phe
of nale
al., 1996i Hanedir a al., 1994).
s€vcr.l g6€s loolcd in AzF Egiom ac exp6s.d in rh. t€stis. Reijo rr al,,
lh.r tic d.l.tio6 h dy o!. chreno$m.l qion @uld
(1995) lhor€d
sFftaroSei*is csing infdtiliry in ma, Bddell
chohosone deletions in 9 our of 80 sospemh
AZFC
d.leriotr $eir
t*tiole
er
hd
(1998) d.teted paftial Y-
ar
(l l%), Two pati€nts showcd
biopsy showd hyDosFm.rog.n sis. One plti.nr wirh
d.l.ion of.ll lhE AZF Egim shoaed S.noli C.ll Only ryndomc
&d thc mdning six
T6liold
biopsi6
p.lio$
Evald
nr?o+.m!to8E$is.
(SCO) on biopsy
h.d deletions involvihg AZfb .bne or AzFb rnd AZFc.
@n
sp€malocttic
Bd&ll
a, al.
6sociatcd wilh sp€m6togcnic failue.
(l9E)
in fou pdi.nls
sug351en
Bli th.F @ r@
od on Ba h.vilg
$.r lhc AzFb
delcrion is
AZFC d.l€ted men Rho have
co.c.iv.d multipb cnibr.. natuElly (ChMg .r al., 1999; Oarll .t al'2002)
eN oflh* 6a
c
tI.
al$
deletion show.d a
sho$ld . Dild6
all
ddclion bE{kpoiirs in 4 infehility psri.nrs
*nh AZFa dcletions. All paricnc hrd DFFRY (USP9Y) sd
3 patients
ad
infdtih.
In 1999 S4c.m.r at rc6n d
ed
disNpt
hEd DBY deleted. P.ti.nts
sd.rc
SCO phsotrTc,
olig@ott@ic pboot p..
s
EST,
pM AZFaTl,
aZFsTl d.l.t d
DFFRY,
dd
OBY
wh.r. a lh. pati.nt who eroin.d DBY
2l
Su. .? ar (1999) cofthted sp€@ro8lnic failur.
lintd e.n.
by
s.quacing $c AZF!
sd
of
ba @dd sdadl
litl.
s.qucn@
wid$,
these
tulclional @ns.qud@. Th.y id.ntifi.d
@sFmic tM, lh. nltalion w6
csticul& bioP€y
dcd€d ryab.togaic .!r6t
ch@terialion of nlrudlly
4i. (2001)
chdct .ird
ocui.g
g.n.s in
most of *,ltich
non{bstructiw
For
in
!
a Croup
nol
sainif.o6
of infeftile
od
ro
p.tidt.
d.l.tions in AZFo, Kuoda-kwaguchi r,
Ggion in 48 in&.lile mcn
foud rcha&ably !ni&m del.lim, speni.g a 3,5-Mb seghdt dd
$@ly olig6Fmic
eG
(lds lh& 5 nillion s?.m
N sube.r6 for hmologous
MFc
delerioG
&d
F.
ml).
bounded by
lambin.tio.
D.ldions of the AZFC Egion of th. Y chtuno$mc
krcM euc! of rFmatogflic
.
tubuLs of lh.
They
thar
'.
bro0er,
re .aFhic
229$ dir6r EF4r
b. heitlbl.
f.niL
in ni3
who
o.
Y-
$o
appai.d
pqdl
a
idatfied
They
point nuhtioD in USP9Y
ifiGtitial Yq dcl.rioN of AZrc
thc
poi.t nut!1ion in
rqid ofdic Y chrcnoehq
tundioMl g.nd US?9Y &d DBY. ScEding ot
f.rtile
10 6
e
lh.
mo3t
Mmo
failure. To d.tefrine the incidcoe of vdious pankl
rh€n .flccr on
fdili9,
K@da-Kawaguchi
.t al. <2001)
dcr.mi&d thc @nplctc nucl@iide s.q!.n€ of AZFC by idenrirying 4d
distiogonbi.S b.lwecn the
&
idcnti..ldpli@N (ldge Ep€aiedieque.@
irddtive mpping-eq@cing
ftom ra.dem duplietion
compkx .onniis
ciei
p!DB. Th& idotisl Epets
dd inv6ion
eenc
may
blocks)
NiiS
h.v. cult d
during pnmte cvoluiion. Thc p.lindonic
fMilies all exp$Fd
in
tsti.uld
thsue.
28
I<Eue ,r al. (200D .valur€d th. incidene oI Y .hrodosome mjcrodeletions
in
!
@nrrol ru1. Fprlation
wh€th€r
sd irfrflil.
m.l.s of $e Deish popul.rio.. To
th. suboptimal sem.n qudliry rcporrcd in
in i.Lr@ of Y mic'odeLtioG wirh
rsFt
to
Ddm{k
pdidtl *king Ift&ttopLshic St m l.jdion
f.nile
Fens md
det cted in
dltated
I
? %
(ICSD
paidls hryirS sFn @wt of moE
an/cr'TtoesFmh. n.y foud
sFificrlly @iat€d
wirh s€!@
di.d
nlltmol
out nudy
100
nl)lml
genenl popul.tion. No d.l.tions
<Ll.tioN in idiopdnic @o/c.t?toz@spedia
wih mnidiolslhi.
e
rilnlry p.Ms tom
nffiGpfric
ssialcd wid a I'iehq
oth.r popol.rioG, $ey
on 138
107
is
ddi.e
ds
dd
wE
I x ld/nl. I1Fy
io 7 % of individuah
Y chrcdosom. micro&letions
$dlrogcnic f.il@r ed
lbe
f'lqu.tuy of Yq
microdeletio.r in the Ddish populatior is 3inild to Ihal ofother @unhi*. Howevef
Knw .r ,.i. found ilc hlploqF .ssi|tion in .bov. oario..d
idiopadic
oligomp€mi.
Thcy lound r hiSh
or
i!.id.M
ras?emia with
ihai of
of h.plogoup 26 (27,9"/.)
lic
$
grcu! hovins
homal conEol popul.tion.
comPared lo conrol
Deisn
mrl. pQuldion (4.6%)- Tlty hyPod6iz.d thal this cl6s of Y chonoeo. m.y b.
ftk
d
for inf.rtility mong DMhh m.n.
The AZF., AZlb, and AZFo intcry.ls were dcfined by incslilial Y-
chod@ne d.l.tions
tl[t
impri. or atringuish sP.mttogd6h R.tpirS
./
ar
(2002) studi.d mme of the legcsi of all hume inreditial dclctions for which deletion
junctiotu
ed
distind AZFb
conpl€t€ int d.ning sequencc w.r€ avsihbl.. They conclud.d lhal no
ilt dll
exins, ltlhough
ilc
AZFb
od AZFo e8iotr
nad
hen lhousnt
29
to b.
non{rcd$ping (VoB
ffi
widts (Sl$)
fct@ds .r at
.? aL, 1996).
slFoxioiLly I
ofr[.
2003 6ed single-Dlcleolid.
$qua€
id.niry
17
AZFC
h.plotypq md h.v. exmin d th.m for d.l.tion of individual AzIo gdc copi6.
ftey
lound
fiv. individuh who l&k d SNv3 tom
DAZ3IDAZ4
urltss
DNA
ad
egns!
b.ldged
lack of |h4e SNVS
Tt?ing I lE biD!ry Y
a larye
egi.nt
o4k6
to
of DNA cont i.ing
wd due !o d.letio. of lh.
sftosed thar
R.tping
?t al. (2004)
a
Enov4 hdf of th. AZFc Egion inctuding
8a. f.nilies. Deldcd
un.lerlying
.ll fiv. individuls
identili.d
simild
12
ItF
belonS€d
p.Ms
15
d.lelios
of
nenb.6 of 8 testie
geos includ. RSlvtY, BPY2, DAZ, CDYI, PRY,
ale
CSPG,ILY, COLCA2LY, TTTY3, TTTY4, TTTY5, TTTYC .rld TTTYIT Tney
fowd tn. d.btioi
Fiiarilt
in
bNch N
.hMo$n6 cei.d lh. d.laion.
wid6ppd
in Euope
$.
AZFC. SoudEn blot
sinild d.lclions @c .le obsded in a grcup of
to hlplogreup N.
.8-Mb) thlr
spFific
lh.
shoB.d that
AZFC
ga. doubl.t in disEl
BPv2.2/BPY2,3
to Y hoplogrclp N,
(|
lvlb
ii
th. Y{hrcnosome genqlory,
ii
which all
As h.PlogrcuP N G tnown 1o b. .ommon
dd Asi., {d
theG i5 no indi.dion of Educcd f.ftilaty
6d
i. sm
wi& ihis Y chrmoema so ihoy @ncludcd thsl s coinon vlriaht of thc human Y
ch@md. lekr
ftniliiy. Lu C
,
/oligozoospemi.
dislal AZfc Egiod cnd th6
ar
@00?)
!$d
ed 5r5 f.nile
tls
eqa cmot b. Equitld
ml.
285 idiop.ihic infertil. m!1. individuah will] @o_
rfl
iD
otdd to exPloft lhc ElationshiP benreen Y
chDd@nc hlptoSroup ed sp.matogoic liilud- Th.y $cd a s.r of
narkd 4d rubjets (cdes and co.Fols) fon
Thay
for
fohd signinor diF.MG b€5v*n
thc
the sam. Ggion
n
quencics of
$.
12 binarv
wq
hg
('
in
$c infedil.
t0
ml6 ed mm.l @tol pot{htid. Th$
ofm.l. in&.tility
in the
SIR) tllele duplicltions
lb*
bd !
dsk flctor
He Chin.s.
Boeh md Joblinc (2003)
carryinS
Y hallogouFK. midt
witti!
duplicaied
det€cted
Y{honosomal short bnd.n ftp€r (Y.
lhc AZFa Ggior &d showd lh.l 2 chono$nc3
6llcl6
conr.in
d a tnird juncrion-specific
Eroviil chmdrs (HER9 s.qu@ [email protected] l'sis of ttre
endoge.ous
c!s.s, shN.d
dlll
bFltpoinls lie in th. sm. Egion of inLFHERv equ.nc. id.ntny, is do deleion
ftip@rl
of lhc
ih€ [email protected]
folod
bskpoinE, sugg*ling lnat dr. ft6heb6 of dullic{rion is th.
n chsis
To
in
Gulring in deldid.
d.Gmin.
the
E€qM.y of g€.etic dal.tioN within
tri.d
vdiols world populations h.rc bm
inf.nil. od fifty f.nile
lwli
our,
MadSe.t al (2002) s.lged 6l
d.n ld nidod.L.io6 oftlt Y chrmoen i.rclving
thc AZF €gion. Th.y fou.d fricodele$ons in 10.8 P€ocnl
ThG
nd
CAC
oflhe inf.nile subj.oE
apat nmbq rlDvEd ! si8lifidt dill.M@ in inf.dile nei
conpdld sith f.,tile nen (16.6). Among ?00 inrenile m.n wnh
6il!r.,
Folin ,r di. (2003) {ound lhat 4
ofgcim..lls
delcdotu
ih
tn.n tesls dd
2 with
uGht d subjdc
sPematoS€nic
(2 with a onPl€r€
$vcF ht?ospmatogch.sir)
(18.6)
abse
nad Partbl AzFb
sld.ppd.nrly idenial beatpoinB. anolhq 8 .fLct d m.n
had @nPlctc
3l
EIA\\Ay.t dl.
@ied out a nudy ro.stablish rhc
p€valencc
of
nicod.lnioG on th. Y clsm@mc in inenil. T!.kbh hal6 wirh .zosFmia
6
(2004)
oli8oespemi!, To d.l,.mine Y chonoson mic.od.l.tions sev.El s€queieljgg.d
Find sc !w nulriplq.d. Micio<leletioN io AZF., AZF1 ad AZF.
@M gdo Fgion! w@ d.lct d in two ofth.60 (3.3%) idiopathic inlenile nrl6
sit
(STS)
eith iomal
b!t!9pd
suge.sting rnal
.
.nd
SRY
8d.tic $|Hing
tlsloc.rion
B
d.t.mined on {6,XX m.1.,
should b. .dvised !o infertil.
ssi$ed Gptoducriv. trEarrn.nts-
!l
^v.dr
er
el.
n.n
b€foE $anins
lz00'4) d.rmiDed
th.
nicrcd.lct@ in 3l i.f.niL tul6 *irn u(h!ccb5l. chon@nd Montli€.
oul of the to$l 33 ( l2 %)
ne. h.d
Yq( I I ) hhrodeletions, lhus supportine lhe
apo.!.d fi8u6 in othd popohtiG.
Thc offiN
4
c.s
had
sequ.nc. deledons with dieEtc hislological findin$ of th.ir
delerion wirhin AZFC
@
hld
t
t$i.s.
Four
avdalc
tr.d.h
Single sY272
w6 rlsincd wirh s.noli c.ll only syndom., wh.@
plti.nr wilh i$l.lad sY84 deleiion wiiltin AZla
maining
singL dMl
Yq
hod
l&ge dcl.tiod in AZFa{
hDorh.sis d1.r th. Yq(l l)
a
imdatuF tesricllr erucluE. Tbc
ed lhod $auG supponin8 lh.
ei@mpss $..lic d€temindts oI slatuE
besides
$nG
@nrrollin! spmdog..esis. Ahhough del.don of tne comPlele AZFC incdal of th. Y
chomo$d.
is
lh. frost @mon
hoq
lpn tic
tus
of hum&
pMi.l AZFo d€lction! (grlg ed bl/b3) tnd Fmov. sma
harc
'@tly
h..n id.nnfi€d
i.
ft.I.
copies
of
infcrtility.
aU
TM
AzFc 8...s
inGnn &d Lnilc popllations by Machev .r 4l
(2004), and an ssocialion study indimLd dat &e Erulting genc dose eduction
12,
Epr@nc ! risk b.tor for sp€matogenic f.il@. Th. onlt deldion
eciltion
e
deletioru
a
with iDf.rtility
lr (!
DAZ3|.!-CDY
= 0.042), $ggesting lhar most
be
!.d
pair
stippcd
e
g./9
n€utFl veidts.
In th€ aZFa Esion t@ tunciionll 8!nes USP9Y
(Ldrn
tyPe to show
Pas€, 1997;
ud DBY hlG beq r.pon€d
Bown ., al., 1998). Sui .t a/ ( 199)
dcndo nut lion in tn€
sd pnt ii uu€tion h
donor sPli@
lh. firsl to E on
a4
d
eion |o be
.dsFmic pltidl
Th. AZFb
sit oi
non{bssudiv€
rd
USPgY €using
Esion coliriB RBMY @did.L gcne (for lFrutogn4is), nutltion of Phioh ha
b6n Gporred to ctus spdabgdic arrd in Dous Th. nrst aZFc @didd' &ne
to be i<Lndlied
ws DAz
by Rcijo
.r dt
altsrimr.lv I in
no$ @nmon, dfeif,g
oftne Azfc inreryd @ &e
095) D€ldio$
I
O
n€n with idioPothi.
'esFmia
(vogt
Sq€ning for Y chromosom€ hicrcd.letions provides cocial infomalion in
rhe
@un*lling of couPles s..kin8 infertiliq
Y
chrchoon' nicrcd'l'rion
s
ofscvoely imPand s!.matogen66 lnprov€m'N 'n
biologi@l achniqB have n.de ditgnosh noE 'dsibl' in Dudn'
scEening is advis.d
noleuld
r.*neni
i!
dalysis. Howete, Y
chonoe6. nicrcdcl.tions de
A*nin.S.ifcr ?t dl., 2004. pFql€d
!
studv to
not
diagos'd
dd.min
reqlic<t for this e.lysis, in Frmcc Micrcd.l.tion detctio!
fast
ed @6i5l.ntly.
tr
lhenfte s.€m n*s$rv for
in all
IvF en&!s
rhe Eguhiory ond'tiots
M
found to be spi"ding
a @nsosus to
& crh'd
on
idi.dd tuI rrgrrr.d. !t!8 tud 6. ir$(ndd cffifu tl|.rbnr
b d ar C.004) cmlcd ib @ @lt ohoi!.d Aoo v3 p.d.d w|& !dob-nds r-.qqnia olirptu r.@on
ri! bfrriL
qH!
frd
E K.
thoM io h.v.
h odholr
([email protected] 2mt
dir a olitoro..prnir. 6.Ot of
Disodddc
in
A2lc Fdolt,
r,h!. 6 c6
CHAPIIR
I
MATERIALS AND METHODS
3.1
IDENTIFICA-TTON
AIID CEARACIERIZATION OF
NEW
IIITCROSATEIJXITS MAR(ERS
J.l.l ldot'rli.ltio! olNr Mi.i@tem.
Y{brco(,!@rl DNA s.qu.@ of
wliteb.ad Ilstirur./
s.q.wi.Eitedd
wT ccene
l-E6 Mb
s $E.o.d uing dd! ton
scq@cing Prcj€ct's dalsb.s. (htlp://M_
publi.-Fl4€/hu&Y. 3ht!l) s d*cnbed
clo!6 tbrt s@ scre&d
60r
the
th. iddti6catid of
bv
w
Attb .r
't
2000 The
Eicrs.Luitc islud.d
ACO0582O, ACOO44?4, AC002531, AC0O48I0, ACO04677, AC0O2992, ACSJ4777,
ACOO5O9, AC00OO2I, AC000022, ACOO6565, AC005942, AC006371. AC0069E?
ACO072?5. Ite@ clon6
d@ibcd by thd
wrc e@ncd by th. prcg1m
BcN! ir 199,
[email protected]. Micrcelellites
vd
nicmslt lliics wE nalc
sPeci6c
gnoDc olhd
tbe
Y
$6ich
chsn6oo..
T@ded R.Fai finder
h.lF in tDdirg thc epets i! th. givc!
choh ftoE
s ih.y.lo
the
ouFd of this ptocru The *l4cd
not hav.
Seq@d@s
wE
a'ni.F
lleir exet Eatch
lheu
!!t i!
lw,ceil.
Foslrd Prinq of acn sd
dultipLd E&tion. Unhbelled piinqs md lab.led primes (Table
Mwc
olisp (Biotech, UK).
in lhe
rle PoEu Prioq 3,
this softw. helps i! Pnher d6irlljrg /hltb://w-eenonc
bir/dipd/din€rl
&d
6
3
ei nit.educqi-
Lbel.d lo hlvc
i) rcrc suppli'd bv
l.1l
Ch.r.cr.riatio! ofNa Mi@!t lUtd b P.tnbnl PoDoLlio!3
Eogiftqi4
IBftn of Bioocdical tnd G4tic
oBCE) wF ed for tte YSIR typins, th€* wples *t coll€cLd
6 t p.n
b@d
Population
3.1
of
Mpl6
ed debil
aEilable
!t
tIE
e.none div.nity Foj.ci &ob
is grv.tr in
vdiou Egios d6ctib.d
in
fis
Appddix L
!.1:.1 M.ldtht PCR
For YSTR typilg of
E&!
B
sph B
O. Palilrdi popdaiiotr hultiPlcx
dplifi.d
for 5
ba*d (!dds
r.beledeith eil,\q of IET, HEX
d
P.i6), o!.
FAM on 5'
l.b.l! fd ih. pdneB. Ttrec o.*eB
PCR lechniq!€
Gld
pihd
wa ued.
of c&h
pln
Il s etn flo@st
(DYS6a8, DYS649 3!d DYS650)
*@
tcE.|@l@ti& Epels, @c p66 !.ucl.otftle Ep.!t @YS652) ald oE h.ra
neeLoiid. Ep.at prin.! (DYs65l).
10
X
pind
prcEix
ws Ptp@d
usins thc
@dtlio! d descnb€d i! Tlbl€ 3-2, lrd stoGd ai -20 'C Th. dultitlcx
tcR ssay M perfomcd i! l0 d offinal volue (S@ Atex II). Th. emtion
6nrl
cosisr.d of 0E 2oo tM .lNI?s, 0.25 U of Sl4€r T.q GIT Biot*hlologv Ll4
cebddsc), 0.083
Fg
Tlq Stut Artibody (Clort ch lrb-
uM ofth. YSTR. MgCl2
m
supplmmftd to
Sive a
Ilc
Japd)
ud
01_02
f]!al concentration of 3 nM
E pc-irculstcd at Mn tcdp@rd for 5'? oil aft.r
in€ubatioo ltF etibody @ added to mast€r dix ad disp€sed inio tub.s
@nrlidng 20 tg ofgcnomn DNA PCR Aaplifiqtion M c.ried oui bv ainS
ib. rouch doM PcR Poto@l Folo@ls s d€snb.d bv Jin .t al l99?. Tlt
Thc Ta4 Sra.t dribody
J6
cdi.g pam.tcs wce pr.-ir6b.tion for l0 din
&aan6td.t
'C
94 "C
fo. I Eia '..eling !r 60 oC for I nin lrd dicBion !i ?2
tb. 661 8 clcl.s, Thcs 8 cycl6
'C for
5
nir
3lD&'.4
Thc
w m
@plimr of
0.3
on
d
E
cn 4
br on
7o
denaturing
Ddr
wr.
fltt
collccled
cxtecio!
2.4 Fl of
tullowing
at 94'C for 2
Ccl
s
uy B.l to gll
vdialion.
corespoldtr. bel\r.cn
(ThoE.!
er
the
ai., 1999i Ayub
F*
s
2
ei!
i
e
cE
l le
b€forc l@diig on
cLcttopLoris.d fbt
eft*e
!*
siz. stu<ltrd d
.ll.l€s
4l , 2000).
ths
and
o) IrE su.l. siz.!
}w .ls
nmb.r ofEP.al uit
er
wlil
at 72
l
5
be!Ac$6h iFt udioa.
c.libEt d b6.d upon th. nMber ofrc!.al uits A
u!.d
4 'C
o$
cdioilg foMide
loding dvc
uirg th. ABI @[6tio!
wE
*a 6i.d
9@ l€pt ti
.stimi.d uing GENESCAN softwe (v.6id 2l)
MAKI33 ald MAKI36
ctcL for
oix.d viib 0.3 d of TAMRAI5o $tich
we d.4ted
uhsCENOTYPER softwaG (vslior
ir.4h
ABI 377 $ql|.r@ cing
polyacryldid. g.l nD
AlI l77 DNA s.q@6
oC
fono*ld by l0 cycl.s of 94 'C fd I
tLc s@pLs
&
!e<16 intcmal sie $.8ldd.td
The sanpl$
Edecd by 0.5
Ein. A
L
Aftd conpletion ofrcR,
od d.xtid blu.,
12
wF th6
for I bin aod 72 'C for
PCR products
E
6
fd I oir 'I!e @.aling tcEFElw
bi4 56'C
'c, fon@d by
ar 94
F68:Mt 3i6
th. dl.les wa. $otd
ofaU Lle ldPles
of DNA
*h
w€
wE
splcs, MAKI32,
Cpl
in odd lo
@!r*t
Fq@@d to d.v.lop
dnd thc
allel. sia
l!'
i! be p'is
3E
Trtl. 3.1 YS'IR Drte6 &d tbcn r.gulc,
P@l cdililias .U th.!.
@t6
d!$sur.d $ YsTRl.
Dv.
UrL
Sn nld
Op) cdc
(lM)
DYS54E F
CGACCATAAGGCTOCAGTT FAM
DYS64E R
OAACICACAT{GAOTICICCTCA
DYS649 F
TGACATCTITGTOOCAATCTG'TET
DYS649 R
OOTCAGAOTOCAAG
DYS65O F
ICACAT(rcCTACAGTCACAG€
DYS65O R
CTCTTTCTCCCTT@CACCT
DYS65I F
CTCCIAGAGAGGGGCAITTCAC
DYS55I R
A.ATANATCACICIC,OAGGCATTT
DYS652 F
CACAGCATGCCTTGGIITTA
DYS652 R
TTTGCGTIATCTCT(rcCTTIC
CATT
250
0.1
0.t
ITIC
lt6
0.2
0.2
2EE
OI
0.1
ACCCAA
304
0.1
0.1
M
0.1
0.1
.t9
r.l.rl
Sbrirti..l
turlFit
AI€b a.q@ies wcE crlcDt.r.d for @h 6&k r Eilg thc Mimon
Excel
sonwa @d
thca hlplotypB
hct€orygosiiics in populdtion
3.2 ESTTMATION OF
we @ rrELd ltultotype fr.qlmis
@pLs w@
abo cd.utlted in lhe Micrcloft
IEE M[IT TION RATE IN
Ald
Exet
CROWING CELL
LINES
J.2.1 SuIL Co[adotr
Mondu populatior sdples avail.ble
at IBGE
muradon Ete in thc gbwirS ceU lines.
P@ urcd &r
FBh
bl@d smpt€s wi{rin 24 lus of
collstion wtc Bed for rbc rrajsfodation by EFLin Bai
(loml)
M
ofth.
the .srioarion
viru
(EBv). Blood
coU*ied in veulaitur tub$ co,r.irjne lcid ci!a& d.xros. (aCD).
EPst
i! Ba ViN *rs
lD.r
to
pnvid.
cls
us.d
0o
r.rsfod
th€
B c.lls to
cmr. tynphoblsioid ell
for g@tic &dysis.
3.2,2 EEv T..4forErlion
Thc blood sdple of5
th. falcon
tn
nl ws g€ltly lay.cd
on 3
tut dd €ntjtug.d for 20 Dinul..
nl offiool h}?.qE
The butrycu
i!
15
nl of
6r€
inrc.f&e of plsma and bisroplqw. Th. Iayer conainitrg lymphdrld
w*
,o
llGftftd io l5 ml &lcoa tub. conilinin8 l0 Dr e!s[ n did [Prtzl Ps*
Moodal ristirur. (RrMl
)1640
Mbirins
r% Fehl cslf
stu
(Fcs) ald l%
Clullmia Ptnv.t Pdeciuin .ud Sr€ptoDycin (CPPS), nix.d wll srd Ftl.ted
by
c.
rifteation @lom rpE for l0 mh. Washing ir <lorc
s
Fi@ll
wb.d
it is ioxic io
rqic. wirh
[email protected]
5
all.
Th. suFrurani
nl *rsn D.did.
dnl th. cdl viabiliry
Ancr
B
ro
mov. dy tri..! of
ru di*riH
Bhilg,
b.s@d
2.3x loo
rld r!. Flld
s
elb wE
fd
Bed
by thc h}?a! blE cxctuion r€st
a'ns henoclroEdd Gtu! 19E4.
Ltrpheycs
9w ta3fu
(o 25
rrqtlf()@rio! ucdiu cortainilg RpMt-
f€ol calf
!.@
C
or cultui!8
o.diu
sulplch€
su!.@rei (l Et) ws .rtr.d ,!d
iI [email protected] rh6ph@
od cE*fod
t64O
ed with
(ICS), 5x105 Mold b€L-ncrcer,to.lbool
cycldporia A. EBV
3r
6 q E fsk @trrliling 5bt of
(19E7). EBv
of 95
%.jr ald
i@dLliad eu
rhc c.th
5 % CO2
could b.
s
cpps, l0
ed
*@
0.S ug
yo
/nl
[email protected] at
drsibcd by w.lt5
visuliz.d.td
l-2
*!cls
s &.. lloadng roud clups of {r. elb. Ilcrascd acidity of the
b rllo
e
indicatio! ofrh. cels rE4fomarion.
oditond Fgoldlt ui.8 i!rcncd di@3coF rd
n.di@ a Equind. Osc lh. c.lt lin ! w€
94
cstlbtish.d
Ttufom.d c. s 1@
srpptcn .d
t!.y qw
wirh
iqh
split in [o rhe
3.2.3 C.llCoutiig
CcU
CoudiS ws dorc Egdaty b calcuht ih. dolbting lin. for th. ells. tOpl
of ttF
cdtr nediu E
irten in 95
et! pht ed oL.d
wilh eqld
votu. of
d'.
0.6 %
trypd blE solurion .ad lo.d.d
h.D@yto6ct
r-
th. neilod of
l(lw
C.U5
EE
alo$Ed lo
d
$nL
thc
d6
tur a f.w
c@rirS chdb€r of rh.
s@ds bcfoc coudng by
atd Pandson (1973).
3,2.4 DNA PREPAX TION
32.4.1 DNA Pr.pntrio! hod Btood
DNA
%
Apprcib|tdy
5
cx!ac!.d by F@cdw
dl of
blood eEple
v&!l!iM- ftFc
dm.!
EDTA) wB added
b rh. 5 nl
volu.
of
B
e[
d*tib.d
by
Suboot.,
aa, t9E9.
@U..cd in &id cillri. d€xros. (ACD)
lysis bufer (KHCO3, NH4CL, &d 0.5 M
w
nir
Suples ee c. ritug.d st l20O rPE for l0 biD .r 4 .C. SuFm!1.lr B
dildLd lrd Fll.l ru GsFDd.d i! diriob vot@c of rh. @[ Irsis buf.r
Elch @pl. B washcd wi€ by lO sl of ccu lrsis b{t€r. To eeh pelbi of
ccUs, 4.75
hl of Sodim Tri EDTA
sulph.& (SDS).ad losl
iEuborcd
ovmidi
Ndr
ph.!ol,
of btmd 6mplc
in
shat€n for l0 Einutes
@uiIrgld !r
3200
K (20 os/Dt)
st ling wcr blrh
rpo for
30
sE
ad
placed on
(STE) butrq, 250 Fl of l0
ofptu!.ite
day the @ples
ed
{
5J
ks
Ms
sodiM dod.lyl
sdd.d &tr
@pls e@
rclMc of equilibilr.d
tepr o! ic. for t0 ninqtcs.
oir !r 4.C. A{hE l.yd
chlo@fom-isodylalcohol @a:l)
for 30
.C.
extricted with equat
sinS cnt tip into eledr. hb.lcd tub.. Io
o/o
ia
6h
SdpL
tuh€s
s @illy
*@
colsied
smpLs equl volMc of cnilcd
.dded, nixed
dd
k.pt or ice ror l0
42
nilut t. Suph olcs we enEifig.d
layd
B
olatcd 6i!g
To
cur dD inio
*t€ste
pot.i@e-K
(20
ng/ol) w& dded
Ein n 4'C.
AquG
E
ho6.
.dd.d. S@d6
.!d
250 pl of 10 % SDS
*4
10
lrl
sch sdple ed imub.tcd in wat r blrh
!o
sh,Io .r 55 'C lior I hou. Sdplcs wE
2
30
labcl.d tub..
th. @pl. l0 Fl of RNe (10 ns/ol)
ilcub.r.d i! sicr ball sblte. .t 37 C for
of
rpo for
sr 3200
rxr&r.d
wirh phdot &d
chl@fod.
i$dtlalolol (2al) b t!w. @irir8 proi.iB. Tho b th. !q@u Lrq
500 pl of l0 M tuonim &ct.re and 5 Dl of chjlled iropopeol (or lo ml of
vw rbrld urit DNA
peipillle 9@ visibL 6 qhir. ihr€d!. Sdpt* *w ph..d ar -20.C fd
chin.d ab$lut..rh.!ol) w@ .dded Sdpt. rubes
ovoignt, N*r dry thc ample
'C.
Sult@tat
w
*ith 5 El ofchillcd
tubes
wF
c@rifi)ged .r
dieardd.!d d€ p.U.r
70 %
elt@t
for 3200
6!!Fd.d.
Do fd
discad.d ed !.U.ts w.r. rlried, Dri€d tupl€s
scl
(pH 8.0, volMe ofTds !dd.d
w
Tle
rtufom.d Itrph@tre
6te in gowing cell lincs,
IE
*E
High
Sepl6 EE wN[.d
nb r 4.C. SuFrid.lt
E,sup.trded in
to rhe
0o
r/
nM Tris
cdcutft. outltio.
o!. molrh ofgrcsah
B
callcd
s
Low
drac&d .Iier llEe nonlh of
PofiL (H!) grcwd. DNA
usins ploced@ descdb.d by Smhook
tO
M
si& ofpeud).
u$d tor DNA *rractioD
6s1 batch after
clild s
rpm for 60 DiD at 4
tyrpioqr.!
Pofile (LP) Crcwth sld 2d b{ch_of DNA
continuoE grc*th md
40
\@
@ddils
U.4,2 DNAdtnctio! lroD rr.nrfor.d
32OO
dt. (1989)
6
s
.xr@te{i
in ihc prclious s.criod.
1
Ccll FUet @nbining 5xlo
(33.3
8.0)
r'r ofl M N!cl,
50
lO
I Etls Pw vorr.ied.rd l9 ol of SIE buffd
nl of lM T.is, pg
Ps &!dd folow.d by t 6l of lb. l0
vie 6d
b6th at
ofgoicirBe K
20 ul
5t C.
%
Thc
6d2 nl
sodiu
dodysyl
[email protected] with
l4pd rqu6N
6tio).
volh.
of 2 M
lth&ol
or
e
Digbl at -20
Tt
'C-
volM.
ft.
blfq
TE
of i$pmp€nol to
Ptripit L
@pLs wE @irifrEed .r
B
add.d (10
incubated at 37 "C for
add.d
dd lh.
tuov. RNe
tube
50
I
br
wiih
El of l0 y!
i@ub6ied at 55 oC ovmight
butrq rh.
spl.
in.
e
FlLicd
and rh.
ih. DNA ws
of chtorofom: isdoyt
tr.lred with
rh. DNA
35OO
oe
knrh
,n
r 3toriDg
ov*
rpo for 30 Eilures !r 4'C
v&uu dti.d ald lbl
rhis t 0 ut of
RNe
SDS
&d
5
d
of prct
of Tris
m
(long/ut) ru
rctatory *Brd bsrh.
i|@ K els !dd.d &d
An !.dding 4 nl ofthc IE
d.qul
votM. of ph.ml lnd
with cl{obfotu: iso@ylalcbol. The uppd lay.r wftn ih. DNA
*ith
.qurl (20 bl) of
incubar.d at 3? o€ wiih .hating fo. 2 hs. To
ws again.xr.cted qith
de ldth ofils [email protected] M
bt&y wde!
bM Tris, I oM EDTA pH 8.0 ). 'IlE h!b.
sb,IiIg. And
qls tunlq
u
ai
by 2 volMe of EBhly diltiled absolur.
to pdl.t rhc DNA. The DNA prll.rs v€re ihcn
EDTA
E
DNA @ftahirg eluti@
ediM e.tltc follo*rd
.qual
$lpbd. (sDs) drcp
phase @nrdring
q.frlly eiov.d ed ext a.t!d wirh m equ.l volu.
dcohol (24;l
of0.5 M EDTMit rpH
ng /ml) &d incubated ovhigltt
spl6 vw
Ncxt d.y
plEel OH 8).
.quilibEred
(20
7.5
sodi@ acebt€
la.dd.d.
{4
Enov.d &d
TIE DNA wN pEcipitated
.qurl vol@c of i$pop€lol ad kcpr w.r rig[t
ar -20
"c_ Th. DNA Ms
by c.ntritug.tion for 30 nin ar 35OO rpd rnd wshcd wnh ?O %
6El DNA p€lla @ Esspend.d in
lO
thcn
oM TneECt, pH
8.0.
.th&ol
3.2.:lJ Etrturtior ol DN
h oriLr lo qlcuLrc tb. c@@tt tid
of DNA, opti..l de.sity (OD) of rh.
@pl.s
B d6ucd ai 260 m ed 2E0 @ Bilg sp4trophotoo.t r. DNA aFcrtrr{o!
M
calculaLd
Ab6o6dc.
Nilg rh. folowilg fodula:
at 260
M
x 50 x dilution fdtor
- DNA con€nration h
pg
/ml
M als checr.d ud @
toD 1.7 io 2, *dich ! fd p@ DNA. Tb. DNA spl6 w th.r !rd!d.t4'C
WIFG 50
i,
corcclion faatoi
llE nrio
4260: /{280
32.5 Sird.prq PCR for SliD!trg. D.l.clion
'Ihe o.thodology .pplied was ,inslcplding so
Dplifiqtion, w.
€r
dr
(1995) to
ed
1% DMSO in
i!.
PCR
.r
to r.ducc th. rcn-specific
@cr ni,lrw s [email protected]
by S.io
.linilaie ihc chl@s of outtipl. bed utifa.ts ald for lh.
c@fi@tion of slipplg.. wli.a N En of tb. Ea€.nrs wE s.ne d ir Dultiptcx
.xc.pt ibi! Eactio! Dixtue wat wiihoui &tibody dd co .iD.d ody onc pdn.r
pair Sirgl.pLr
d!$ribed
PCR
pwiNly
@doN
for
M
.lso
tuid
oullipld @tion
( Jin
our by a
'torchdoh" prorcol
., al, l9?; Arlb
c, at., 2ooo).
s
45
3.2.6
H
ElelDpho61|
w tb.! datopt@.<l o! 4 % &lyboide rek G
ddcrib.d in tb. Fai@ clid for MultipLx |*riod M Sdpl€ aw lqdql
ADplifi€d ?CR prodet
in
{L@& lar.r
da.cli@ of
!o prcv.at
ilipFgc
ay cttu.
of
Aillovq Aon
Tb. @pLs stowirS rlipFge
ldjMt wlh
*!E
E4an d
md for tbc
tlo d ttE
riEc. io @!fi@ l[c Fsult!,
3.2.? Strtbli.rl^r.t6i!
Mutdid 6ie! qw *dD.r..l .i th.
divili@
by tlE
lubd
of
old.tioB divid.d
by
t!. IMhd of
nctlod ofKuict .r a/. (199).
t"= t(trS8
Whc
S is tbc
Mber
of
!.!@r. lci
rll.16 obsd.d for al lo.i lrd n i!
!3
t st
4 X is tbc 106l nuba
thc poputatio!
nmba ed
g
i!
lor
of
qd.!r
s@bq of
MALE INI'ERTILITY
3.3.1 Sd!'L Collelio!
I! oi&r
ad.d
io
detedic rh. ssidrion of infcnility .trd Y cbroEo!@.
5l ildividu.ls &d gFup.d dEE 6 ob.nucrivc
duc ro sp€maiogenic
It
msl, rcvd.
nmb.r of irdivi&Els i!
oligospc@ia and
eh s4p
i5
'@GFdia
oliSonmal
slFM ir TaUc
3.2.
s
tbsF@ir
but stil
bfeniL,
hdividurls
EE
Dirlt b. de
grcupcd
s .dsF@ic $,lr.r lb.t h.v. no !p.d
to obsiruction or sp€miogcnic
pams hwirg lFm coul
<5
oilior/!
@st.
Sdd
&.odi!g
in
sei\
oligosFmic soup of
to tb. WUO
diidia wtiL
irfrlil.
Blood
colLcrio! profotu. ceying all ihc infod.tion alout [email protected] pbfiL, a!e,
.$!c
mMlt (Amx-lID.
l0 Dl
th. forth soup i! lavirg sFtu co6t
b&kgroud wet
of the
v@N
also
blood is
uord.d
nitlior/ ol but
5-20
slong with thc infomed
@lbr.d id v&!1ain6 ed DNA
B
.xt!&rcd Nins
dlia
d.sib.d ph@l chlorcf@ .xtlction Falod
332 S..cri!g
Y
rll
of
chsoosdrl
is
dcledoq s,ltich
.r./.,
DAZ obtoined
sothd s.t
@oE
a
of
nrlpcd uirg th. $l of
MLd
by
lioi
ioponart
!o
c
e
dd
p6is
c Egions, For th€
.letstion
nl'
th€ grlgr
*a
ued to
lrc aoE u&a
DBY ald
cl.{ picn!. of the
of DBY genc in thc Azfa reei@
AzFc egron dclcdo6.
anplificatiotr and prodat size
b
dlso used io givc a
mplry . s.go.
rid6 sFuilg
e4*ry ClG &r lp@rioeesJs
b 5 prioq
tn. dala bas v@
5
lh. R!i@, ar €003} Iris s.r of
01084/01065
Gw @pi6 of th.
2004). In.ddition
d.Ltiotr pshetu. DBY
DAz n
lw
6ed io Eap U. g$s deletion h AZI!,
of sicrd.letioB
(Ma.hee
dclcrioB
dd c r.8i6 d.stih.d
rhc AzFa" b
deker
AZf R.gioa fo. d.Liion
Ajlprn
giva ii TabL 3.l.
ed
E .long tn.n Egior of
T.tL:l-t:
T.U. .bowils tn. d!!ibdi@ of @pl6
dislcd codnbd
Olol4l
Ot ltudirc AaoAdDi.
Orsq2
AroorF@i. du. !o lp@at%.nic
Od+l
[email protected].
Orc{p
4
Nom.l boligoqc@ia
.r&6i
i!
8ru{c b.!.<l @
17
4E
ftDh
33t
M!rt6
to Dlp tb.
cobd.t
AZF^'ft'-2 AZfa 220bp
or p.nirr dddioG
DBY
SYl27
AzFr 303 b!
AZFb n4Up
5E.C
6.C
5E.C
SYll4
AEIt
56.C
SY254
SY25J
AZn. 350 bp
Azx. t25tp
DAz
SYl532
ALF.
SRY
301bp
bp
l67bp
96
58.C
58qC
ID
5E"C
oflh! Azla,b &d
Rrio ,,
c
ar.,
2003
Omae
R!ic! cl
2@3
Rlicu
.r
ar.,
41,,
2003
.,
2@3
R{6 .t
Rdcu
a1,,
d.,
2001
G@8.
R
io ., zl,
2003
EIE
CDYI
DAzl 923W
An.
AEc l4l
60.C
M!(l.! .t
al-,
2@4
60pc
M&f
2004
et
dt.,
49
w doG foi both prio6 oflhe AzFr egiotr pbx-2 dd
DBY a both [.v. wid. difeff. n lh. &Edilg r.bFEt'E 58 'c ior For-2
siagLplcx PCR
rd 66'c fo
DBY. For
i!. mplific.tion oftE
AZFb Ggion nultidex
aing ulabelled sYl27 tud sYl34 e both hav€
t!.
vbldird bt clhidiM brcnide strining
clcctrophoeed for up to lbr dt
sc .r8dirg t bFnte
*w.aily
dplifiotior
siz. for DAz
B
96
uilg prid6
of 58 'C
viiualized o! 2%
WhiL for the
on th. 2 %
100 volts to havc
Multiplqing a.l!o .loE by
thar
amaling t mpentw and
b.twe! th. prcdEr siz. is 25 ba!. pqia
ditrc@ce
!iz.r
@.
lxl
96 dod
rbis
a
ag!I$. 8el fte
cled sie
sY255
difccne
Fpolior
ed
caily
g€l
ofth€ buds.
3Y254 both hav.
hrrc eid. diffc!!@
B
th.
h th.i. prodEl
agr@ s€l by clhidiu boDide stahirg.
ofth€ DAZ rouchdoM tCR prcto@l wa
w4
prodet
bs. p.ir.
3,lJ E.plogrcrpilg
H.plogrouF
EF
ssig!.d lo lt€ supl6 by
tyPing
of
tn
si4L
n.Elotide @iario6 (Biallclic odk u) on O. Y cbmosom. and .slablishitrg
or.ir *olutioMry Elaiioship
wE [email protected]
oflh.s
fo. th.n Eos:t .Lriv.d
w. dplifed
14
Mrd 6 sLou in Fig 3.1. tlaplo8ioups
n.nd s &sib.d
b'allclk @kes ald orc ineltior
ro assign ditr€renl hlplogrcups to
ou
by
CYAP)
sMplca. Mdken
v&iou dpli6clrion pl@dG v@ s.d lik ARMS
U!d.Ibi[ .r aa, 197.
dd
onc dcl.tio! (12f2)
@
listed in TabL 3,4,
PCR or RFLPS
*hich h.lP
50
E
t-
srlI
?
I
t
s
i
l!
t'
5l
u
ihc deLctio! of Ihe* EutatioB. I'lE bir!.lic
@plifcrlion 6.thodolos/ ir
8iE i! TaU.
MLd
.loDe wiih Ihci!
3.4.
33J.T ARMS PCR
Aapli6€tion eAeto.y nutatio! sysr.D (axMs)
qcluiEly
on
M52, Ml?o M172, M173, ME9
!d
discnnimrion rcli.!
ile 3' .nd b6. ofr piibe.
Apr
ItPing nethod dd rh€ prioq s.q@nce
2002. Irtehd
ld.
sta'd.rd
6
94 .I
ty!.d
sa desdih.!
only
17,M207,l'124,
tlc Auclc Speifc
PCR
by Und€ftill, 2001 and YCC
Nd i, uplifying ih. l2o
ob$fle !o @plifcadon ci6.r dE to d.Ltio.: or
33J.2
by
M
*t E .lt t.
bdae
of
d.letio!
uplificltior
ro
6ile
FCR-RFLP
VeialioN in ihe Ln81h of esiriciion ftagd.nls th.t @ glnmi.d aftd
di&G asrictio!.EEo (cFdzictd.r dt, t974).
DNA is dis*rcd wirh
RFLPS
iI
de dE lo introdraion or @ovsl of rcstriction .iLs multing i!
thc leDglh of th. n'agncnis
tool to @tutruct derail
dpli6..don
a!e*
Emining
15
RFLPS
d
nosdy EusAl
r'd
s
c
s!
sd.tic linkas. orls (Krysr a al. 2004). And th.
with Hp€€riv.
gel to cn ck
F!du.d.
difeMc.
pde{ et th. l0 d
of thc podrct
@ ru o. 2 7o
dplific.tid of noNFci6c ilgD.nts. Th.& ,ft{
rhc
ld of the piodur wa diSElled wilh 1.5 U of eslricrid @yo.
@ntdning lX EsFctivc boff.r.
52
Trbl. !.a: Biallelic d.rLs
and
ih. !.rhod
Bd
for lheir
tling
io def@
195
tlitrdllvlu
c_c
M9
SRYl532(sR O-A
Y roE3r.D
I
5t4+!94
Undrhiu .' z/., t997
167b9 D6III
55+
I
4l3b!
&l
C
NAOI
A
- C
RPS4YMIIo
C
-T
ll2
bp
lE+295
AUel. soc.i6c PCR
423
All'l' sPeifc
PCR
14u15*234
Bogro
ot
",
,
o*
YCC,2002
ARMS PCPJ NLle
U!&rhill
,
arr000
&2001
MJ2
Ml70
A
A
r C
-c
534
282
bp
All€lc ap'cific PCR
ARr/6 PCR/ Alele
sFcfic PCR
vcc,2002
ARMS PCR
YCC, 2002;
Mg9
C
-T
i,21
Allclc aperllc PCR
YCC.2002
Seqrpr.
53
J3JJ At!ru
G.l
EL.t!!lo6L
AA.r uplifi.ltion 5
d
dngc c,
Ficol
20
Fccd
of th. Orug! O lordios
.rd looDM EDIA
[email protected].
6e *!6ntid) Fd
oflb. lX'I3E Dqft.
39
frlr sdl
n,
rcltagc of
ri4
E
cont
i!i!g
.iLLd io
c!
0.125
tn. bpliEqt
r
ft.
.$r!€.or2rl ilioe@iw:.gtorc gd!(fo!
toililg ed Diri4. TIE
uk
Frcelt
nDpL.
@pL! 3 gnr ofth..g'w s.l ru di$oltd i!
by
USA) or OXI?4 DNA
d}!
lle. trI
disert
100 b?
(([email protected]!, US,{)
FldEt ELotoFosi:
t2cl50 vol! !rin8 Po*q Pacl
DNA Ltlda
3000
w qid
@
150
Dl
(ftm€t
lord.d in the
oa rl .
coad
(Biokd L.bor.loti.t) for 3040
CHAPITR
'
RESULTS AND DISCUSSION
4.I
IDENTIFICAIION AND CEASACTEruZATION OF NEW Y
MICROSATELLTTf,S MATI<ERS
Five @vcl l@i
nucLodde
rFar dd
@istirs of lhe r.6-nu.l@dde
orc
id.
h.turcLorid. EFat wE
silglc tulc<Fcific llodwr. Fou of th. fiE l@i
EF.b,
Th€ p.nt3-Drclatid. Fpe.t
(AATAI)
@
labelcd
for
i!. .!.lFi3
of 756 bal.
ilel., msed
t)mbd
of
n@bcr
or.Iel6 ugilg t!6 ll
AoD 4-12,
lw
bcN
gave a
sioplc horog.M6
FF!t3 (AC005820). Udlg dr!-
PcR @crion
spl6
i6ed, .scn
ws @6pLx. nt Fp€ll seq@.
inEEupr.d by two AAT4AI
pdo*, . linsle nultiplq
EFi' dc Far!-
B
sci up
(App.dix
2)
ad sed
fiod four.n P.}jlrui popol,lios. IlF
Tw ndkeB rce hielly polForptic
(DYS650)
Iig.4.la-e. such lDlrrorphic bdk@
e
to
12 @YE652)
bislry vuiabL atd
with
&d shoh in
e
suit ble for
@pliotio$ ir foeEic $ic@ ieludilg Freniry dcc@i@tion ed bdividuat
ideltifiotion. nth
is abo shoM by th. direBity vslus of tEse
ditr@nt popdtioG
(trbl.4,l.l). Mst..s
vaiabl.
l6i
el@
Pujali populatio! ed
for
DYS649
loM
ad
DYS648 had only
DYS651 had onc
DYS646
t'o comon alLLr
ruiEu
od
DYS649
.nd showd
odkds itr
sE
thc l€asl
ztu div.siry
valrc of 0.56 for Buruho popuLrion
comon allcie vith lh. oder all.l.s @ceing dt
fEque@i6. DYS649 hrvc shom
e
divcsiry valu for Kdhnni &d
55
ft (1.:
Rcpcd
uit filqEsy
or DYSA8
lo@ ir Pttji.a
I
fq 4rb: Rc!.d uit tlqtlrEy ofDYs6a9locs in ttt&br
:tr
!i
rta,rg
R.?.d uit Alqultcy ofDYS550 !@a in P.LisL.
ln
iii
Ft{ld: R4.{ bit ilqtta.y of DYS65I l@ain Ptrilt tl
57
Er
Er
23
naarc
R.t d Dn f!q!@t
ot DYS652
Loq! ir
P.ritu
5E
I.bL ,l.ll: Hd€roz*ositi* .d
Prki$ei
I'STR
[email protected] ofAlLLs for aI five
6 itr
€tbnic grcuDs
DYS6.t8
DYS649
DYS650
0.4E@.@5
0r2l+0.014
{n=8)
0.000drlqt0
0.25010,t24
(n=8)
0.r501{.1.24
oofltro.o0
DYS65l
I
5
0.?3la{.009
DYS652
t2
0..177i0.015
0.@71{_102
0_67914.061
0.{d*!0.135
0.00010.000
0.607+0.102
0.730*0.025
0.432*0.050
0.810+0.012
0.txt0c{.000
0.659r0.082
0.J85+0.104
0.6?040.085
0.505+0.002
0.2r8r4.017
[email protected]
0_10240.041
0.65010.012
(F12)
0J60r{).017.
0.203*0.042
0?J?+0.018
0.1t814.040
0r5lr{.0r2.
(a=21)
0.519*0.005
0,7244r23
0.484+0.034
0.670i0.010
0,t07a0,004
(n=42)
Sindni
o&t
0.J45i0.051
0.13E*0.050
(n=t8)
0.506a{.006
0 220+0.048
0.790*0.0?4r
(Fl8)
0,471a{.052
0 209a0.081
0.595+0.0?4
(!"EE)
0_4E5a{.015
0.25!4.03E
0.761a{_016
0.J84!0.042
0.74410.020
(F83)
0.45140.024
0.09310.030
0.751+0.011
0.6204.n29r
0.76914 0r9
(n=66)
0.41640.01J
0.19E+!,045
0.500!0.050
0.ll?riLtl3t
0.579*0.043
(n=lL7)
0.442!A 021
0158+!.030
0.705+0.021
0.234+0.035
039244.0t2
59
ndiau
Balti populatios
ed
DYS65I 3ho*rd
& [email protected] lewl ofvebrio&
for Kahniri population this
valE ol0-510 fd }trz@ pop'rlitior Th. locu
@kd hs z.F
wirh ciglr
div@iiy
diflcanr.!c16, bu
vile.
Two out of
file
Dar&6 b!v. rnom &D divdity for Kahnnis. 'ftc divGity rdEs in
Fpul.tior previ.L vi.w of lh. doluliory FlalioBhip6 b.te&ca ditrcmt Y
chrcnosodal gsups
@rtcs
toD diffdat .dDic b&kgroud. 'fte diveBity valw of
for
al
the popul.tioB
'fte ovdall
C
vd@ for all th.
6on
wi€d frlD
9% @YS6a9. DYS550) to
lai
N
0.221 (DYS649) ro 0.795 (DYS652).
12,6% and for
t6 % @YS652)
idividu.l
s shoh
@t6
vdi.d
in TaUe 4. L2.
ofl! 6ve Y-STk individual h.plotlTes
l* odtret d Toral 154 t tlotyFs v4 foud in 7J6 @p16 Oa of ilFe,
70 haplotyF3 *e Fcgt h silgl. iulivid8ls, 2E w s[!Ed by tm
iftlividu.ls, ll by 3 individurls ard thc nbbd oI sha€d haplottlB incrt$ed
g6duUy. T@ h,plotype r@ foud dor 6.qu.ndt, thc h!9loq?e
9 8_17_ll_24 la sbrFd by 105 irdividuds tbnt B 13.8 % and9 8 l7 ll 25
By enbini.g th. slhlic stat*
wa
sharcd by 68 individuds (9 %) G
shoM i! Table 4.1.3.
tl.plog!6 de hiShly inf@tiv.
lcsftn nhtioNhiF &d
fi5h.md tib.s, dhibiLd
6
male linelels.
b *lr
they prcvi&
e
icight into fien
Tle Mohrln , qho @ th. &cie
m.ly low dimity of 0.66 tur rh@ fiv.l@i.
similarly Kadmins hav. also shoM low divdity value of 0.64 (Trble 4.1.4).
OE h4bqT. 9-8-l?-ll-24
m.omt
d
@YS548-DYS649-DYS650-DYS65I-DYS652)
fo.57 % ofhis popuiatioL
60
T.bLa.l* tLL@ysdniB.!d cnlrlEbrrll6ED.rt6si!or6ll
LOCI
Grt
o.t25f,
0.52146
DYS64t
0.14912
0.tE535
DYS649
0_09(t
0.22091
DYS650
0.0915
o.7gN
DYS65l
0.1fi7E
0.37645
DYS652
0.1510?
0.?93E1
5l
I.bLa.B:
Hrplotyp.
cmbiEti@.!d
thcn
A.q@y
in
?.I$t&i
DYS6aa_DYS6a9_DYS550_DYS66l_DYS{t
9El?ll24
9a t7 lt25
8817 ll 26
88t?lI23
94t5t227
881?ll25
98lElr,3
9E l7 ll 23
EE rE ll 23
9815 ll 25
9816 lt
98 t5 ll
8 818
24
25
11 25
9818 lt24
ll ll
8Et5|l26
88lE ll 26
9E
2.1
8815 |l25
105
0.6E
68
25
0.44
0.162
25
0.t62
2t
0.t42
19
0.l23.lBd.l8ldhlD
l7
l6
0.110
0.103
0-155
0!97
I5
t5
t2
lt
ll
ll
l0
l0
0.097
0.097
0.077
0.071
0.071
0.07t
0.065
0.065
62
T.bL {.1.* HlplortF H.tozrgdrty
!oP'&rids
{dq
fo. dl lh. loci
E.plotyF E [email protected] v.lft|
0_75
0.61286
0.96727
Bdri
0.75t24
0.90E49
0.9t36
0.81476
0.9E2
0.98
0.E62
0.9t1
0.942
0.66154
Sindli
0.941
i! P*islani
63
'tli.ty eich individulk
our
ofde sixty 3n tttlr&d
u
ftcqEncy of thjs haplotrF
PolDlalioD
@lEibuiiDg lo tb€ higll
Patistan. Thc [email protected] or rcvcl polttot?hic
ofls !.w
Eicmsatellils od thc Ydmoosode
f{)mic ces ed ir th.
w
losibfincs for inv.sdgation of
sndy of sub{t@t'dtg of cloely Elt'l.d popul,.lioB.
rp*ific h4lottlca q@ ds shoM in @lid 5Mi.5 @icd o'n by
Mohrlidin ,, al (2001) i! rtich for
Psi lopd.tioN
hav. shoM
F
16
rukq
baPlotyF
ulalion sp.cific
Population sFcific bnplogTca @rc
als
t[.
Br.huj,
trdd b@d
9-8-l?-tl-24, ni!.t cn ol'n of boDd.Fd .trd
hrplottpe
riD Moh@.
3h@d by 68 individlals,
IAe
dt
19 iDdividuals.
Thb
disvqy
ou of lh€n
of th.d 27
18
haplotyp€
P.th4
m
.d
a::
2E 7
thG
m
% of th€
P.|he Pop'nadd)
th.n hlPlot T. frEqMcv
will otr€s re* po$ibiliti.s for qvestig ioo of fomic 6es.nd
hme
$e
9-8-18.1I -23 shmd by
(20.5 % of th.
of mvcl polFo.phic oictu$Luitc!
ed
ildivi&als
sildbi
wrc BEhui tbn nal$
@oion
re
*tdten
105
iost @@or laplottP. 9-E-l?l l-25
'@ld
Blahui popllation. Th. tbid most
uPon YSIR
obdwed in Siidhi, BEtrli
P.rhd populatiotu. Th€ dosl @mon h|ploq?€ cosbtir'g of
M
klarh &d
iD the studv
of
population sub4lructurc in Palistrn.
Cd,CULATION OF MUTATION IIATE IN EBV TIIANSFORMED
CELL LINES
Dr
6
to th. Irck of &conbinaiion a
hG h1hs, .xc.p1 wh.n mutatio6
sn cdi.s
lhe
sdc Y-S'IR
@u. SEh nulatioc
e e
baPLotvpe
and their
64
a.qrcncy
i! co@otrly
us.d l@i h4 bG{
loc Fr gdmtion Eo.w
4 d/,,
1992;
ofth€ mut'tion Er., YSTR ndkds
Method
ssiior) w€E
Out of 66
(s
dc$ul.d !t appoximat.ly
Huls
,
2
x l0-r p.r
ai.,2001). For thc c.Lllation
dc!.ribed in Table 3.1 in M.t .ial
dd
usc<I
c.ll lims &rlvscd.
2
cll
lirc3 showd @w EutatioB.
b
o@
B obs.d .nd 30 gcnmtiotu rld i! lb. othd rna 90
sddiioN. A ouraiion *tl coBidc&d a a ew F.t with h.idi of il00
flllo@de @it! in ldditiod ro tn rdrc F.t- This [email protected] 30 % of rLe
tolll &d B ob.d.d i! te ccll lifts. h botb c.s peL h.ighI of onsinal au.L
(ot6a!d i! blood splo s, hisb.. the tblt of ew rll.1. (Fig-4.2.1 & lig
422)- II asdts *a biCt EFodEibl. .rd wa so in boln indieiduds in
l0 Epli.nioE. Oo. od.ti@ d tbc DYS650 tcln u@l6rid. &!..I llN B a
lo$ of ts Fpet uits, &d B obsdrd i! ell liD (MHN 06D .nd l0
c.ll li& nutrtid
sca6li6, &d l!@in d sidilr
in
Fl.tiv. h.idt
DYS6'0 locu bls h.t rczygolity of 0.731
Th. sdo.d Dui.tioD,
nucleotide
rFat loc6,
s.mrioB.It.
ru
i
0.009 in
En
90
gcedti6.
Iad
P.Kstrli popuatid,
! giir of o!. ap*i sii !t DYS552 a Frt!
id.riifi.d in dothc. c.lL
L@u DYS652 havc h.crczygolity of
thal is highest ofaU thc
r
fi!.loci
lie (M{N
049)
0.?95 + 0.004
lner 90
i! Paldsid
iestcd. Thc doubliog tidc $ts estindt€d to be
53
@
n
a.l,t
ddE
ffi.d tdd.ne&..|l
lb.r Shri.s 6. fitrd rir. t-. !d. t low ..d Ft d
u|at . of nld l.it h fr!d.d.. DnA r&plo pr.!..d 6d Uood
(B), a@ t&!6r.d !&$ocyi.! (LPt a.d tor&@.d
DYE550
VEFocyb. lnd
ot ..Gd h EAV
3
Dodb
ir
rro$h CIP)
66
rq €J:
nrtdid ot atEd iE EBV t6bded lyDlbtl.lkid ccU
li6.&6 90 sF.lioa Shovi!8 lt. t.sE@I !iz. t * Ft b.lw
.!.n F* dd lMbd of Epet uit i! p@!tlBi.. DNA spL
Dr6552
F!F!d ao6 blood @), non ae{m.d lFph@}rd
h@fffid [email protected] .frd 3 Bonth'r tur ee"r! G1?)
(LP),
t6
25.4 + 3.1 bouB
(o..o + S. D).
It!
douuils tiD.
s,
cdculi.d
by
tl.
@utirg
e[ liB !n r ady 24 hs B sboM i! Fis 4.2.3. T!.
oo&l dLL * wilG E 17 + 0.93 fd DYs650 @d 2:1 * L5 flr DYS552 in r[.
lllobq
of
poptdnioo
eU! i!
3ix
cxbi!.{L sr. oh.qEd 2 dut tioB, @
tlFrt hlvilg s.Flic dit6ity
EFd
g4dc dilq3ity
b.vi!'g
E
olt.'lid nt
olcd.t
of 0.731 .!d
of 0.795 in
a
P*bt ri popuLlid
&cdde.
F
or- 099D
i! *!i.h lb. c.lcuhLd butalior ntc s
DYSI9.
berliG c re blr o*r
ourdoc @
pe4
of
tb. drylicdiou of
6.
?0
t.m n8Lori&
Cl.blc
a.l.l).
th. ,llr(ly coldu.r.d bt
3.19.
l0r
fo!
K|yld.r
rilgl. lo@
ent of 24 bee Fir i!$nid
F
dj{at !.qudc (au Y ., ar., 2000) .!d Oit
EFll dtpli..rio! i! iEFd&t i! .|tly Drcro{alit doltnioi
ts'nB dlo elebi!
tt
Thc
.tui q) g.c6li@ .!d 3,55xl0'/
[email protected]
io
DYS550, r
gold i! DYS6' . p.!r. !@lcoti&
d io b. S.lErlO_r
locu!. Whicb
iD
digi! oflhdr n!d@ E!a8 ty rcptcldoi ,liF.gc
iani@ .!il .l$ provi&! ih. DuLli@ n!.. i! lh.
ed6 F
It6.
o.
bt
rlg tDDugt
rlp.rcd lo!g.l@ gto*lb. Thi! fndiig b i6pdr.!t i! olvilg th. prc6l@ 6r
th.6.rl(.t3 u.d for plylog.nclic udFir .nd fo@ic !ndi6.
er*a
wbich
e
v.lB o
h.vils low divdity
Ft nity !6ting, a |}a. n t ct !e
6i6o6lt.lliE winio|le.
Borb
E*cE
rbrr
EisL.d t!.
Aieof licftq
8dB €p.cidy
in
3pdr.@B ort tioa @ Ptod@
DYS650
&d DYS652 Ddc biC divdiry
P.li.LDi Fpubtic lbE i!dic.Ii!8 vdi$|. Dte of lle
'@!s
Mrcu. gid divdity lrls s b. lh. Bdt of &c@uLtion of ouLlid
v.l@
duiDg lb.
qole ofcvoluliotr
6t
I
I
a
I
z
naa.a!!
of
dfrrd h/og..tl-id cdl [r!. Ft !..1tu ib &n
&rdiry dc rb g{t loliry |b goet in la6 of nnt r of
Sn
cdb
d..6,rry
2a
hr
69
M.t,€d d dr
@000)
odper.d ln. dichocd of sFific g?c of oicbsatelires
i! codiq egios ei& ihlt
rFlts a
or the coding
typ€s of
@dpatld
rFais
kr pt&rt
{*
foud in lh.
nrc|@nde
noFodilg Ggro6 dd foud eLctiv. prc$@
of th.
!o
rcrcodirg oE!. Funlq il
B
mlcd ilat aI
Egios wh.t !s bi
itr th. loFoding
and
h*
cotling rcsion6 (Yinsl.i Dd Fetr8zhq 2003).
nut tio6 w& only obeocd in th. t
tra nuleotide
but not in L!€ heh epelt (DYS650) ucd
te@ ofspe.in *kctiv. pE$e
ir lh.
in codng
dd
peh nNleotide E!.als
ftt
!fu(ly.
Ite
c@ br qplained in
Egio6 .gritut nur.doN.
:I.-} MALE TIT-ERIILITY
,L!.r [email protected]
of AZF
r.gtud
aor
d.l.riou
A nMbd of rt'rdja h.v!
corlation b€treer th.
t
poinr.d out
qrcncy of nicbd.l.tioro
$qu.@ tace.d sn$ GTs) dalyz.d. For.rdplc
s .st blist d
detelcd .d rubd of
rhit th.E is
ifw
@ci&r
only studies
strich idiopathic @osp€mic or oli8o$p.mh palicnt wee ealrze4 the
of STS
wied
(Najusbadi
a lor up
sr
ar
to E0 but FcvdLncc did not increase
1996, stuppia
F.tlin et al. t9g9,Kftvsz.t at.
.t al,
ifnore
STS
TEEfoa only sv.n
STS
ndber
*r ed
1996, Pryor 2t al. 1991 ,\<rcmer et
1999).
ir
al. 1997,
ndt6 l@
ued in thi5 5nrdy which hav. .lso bcd us.d h va.ioN orhd studi.s io srinate
lhe
pEvddc. ofni.Fd.LdoB.
the obwadon ot
Rai! .r
This
al. (2003)
sl.clion of MLer s.Lction
m
sltt
aor a
n
s d6{iH
thn
based
on
cdirl
70
di.gn6is of thc oicrdelelioB th. p@l of STS should.ovd all thc n c.sny
g.ncs GquiEd for sFmatog.rcsis.
Y chtooo$Dal coes idvolv.d
b.ftd
*w
c
Ggi6-
!5.d to
$lla
Two addititui
ed
ro
Orn
of$s. eEn D.rt
de'eti@ for AZFr, b
@td
@v{ {F
i! spcmltog{*is
DBY ir
dr
&d
c
ir tb.
DFFRY,.!d DBY
Wh.c
6
s.M
6dts
of the delerio!
e.lt%d cmi.s AZla
dclctios
pdiiats, Esulis hrrc lhom
dlt
ww
d€Lliotr
cd 6ue olisosF@ic
d.lctitu
w
syd@. i! p.rihs.
h.vitrs deleti@ of AZIa rgiod
*w
nrund
i.
3.9 % of
shoM aociaiion with s!.darog.nic
ud m.m
lbat
s
d.y
kc
hin
faile.
TLcse
h.yirg
t?€ of d.Ltion.
dc inftdne popdstiotr tbd
tb'l
s
*!s
mppcd by Sdg.nt .r al,
or sCO coDdidoo d.padine upor
rl &rltkd. BEndell .r ar. (t998) sugg$cd
nonc bais
lo$ of
lh. prtimt $io elliEd DBY rhow.d a 6ildd otigo&sFmi.
sD.maiog.nio aEst or obstrEtion. AZFa d.LtioN
of
.lrlysis
deletion rh.l
Hdrilr i!
.a3pdt ed biop6y culls 9@ rct .viilable to categorir
AZFb
1160
by S*8dt .r
nryped
eus.d *rroli c.ll otrty (SCO)
ph.rcg?€. nE parisr i! this study wbo
(1999)
STS
Egi@ in 5l irf.njL ildividurls.
3.3). Thc
2 % of ihe infenile population, AZFa
(1999)
in AZFA,
6, firc plriiouly publih.d Yq
hrva shoM thal onlyon Fcon our of rh. 5l
d.k
Bid.
AAa €gion $d DAZ i, AZIc Fgior wE
Esio! (TibL
'Ainm
nosdy
the
idke6
is 2
ce3 oui
AZn d.l.tior h!v.
weE ba5.d on
.ll
d
to d.t cr th. delerion of th€ whol. Ggion
(Rdcu et.i.,2001). AzFc.lelerioB d@ouis for 5.8 %
oft'& infetile popularion
Tt
Fprestarivc rsults @
tiilt
is 3 our ofihe
5l ces 4 shou i!T!bl.4.3.1.
7l
shoM in fig
4.3.1 ,
et at. <2cn5,
for A2Xb dclaiod &d Fig 4,3.2
@ d.ldiors
le
found
n6t tlqrc
ly imlxl€d
i! conrok, Furthci noe
ar (2001) obswed th.t AzFc Esion qhich
ir
l*
tdy. sloM tbli Y cbrodolMc lonr-m oi6<LLdo6
der€cr.d in 6.070 of infcrtiL ocn lod
whtrs
fd AZF. d.l.tios- a{Eta
tu
rtr MFc Esioa
Kurod. Kawglcbi €,
n6t co@only ddei.d
th. Esion
u@sF@ia ecouk for abolt 12 p.rc.lt of th. ca.s of loFbstrutive
azoslmi.
@ied o'n
drd abort
by
6
p.rc.lt in !cvd. oligorp.mia.
[email protected]
dr dl. (2007)
p.@!t
ed Aac Esi@ we &iccicd a
2.6 yo
loE nl. ofd.Lri@
to
a
.ooled
ou
of
sG
WhcE
a i! Fcdt
Ei.!od.l.t@
in th.
snldy
A4b
(7250 of rhc cdB i! (uwir showing
d.tr
Pritiin d.bdoG in azFc Esion cpod.d by pcppilg .r aL @003)
g/Cr deteiid lhlt Movcd spd of 1.6 Mb,
lolrroryliss i!
ln.
AZFC bul
sonc popdarioB
rdue
ihc @py
fti!
dclctio!
a rrlgr dd.ti6 &
@t
nebdof th..i8ht gqn
Gr/cr del€tios \N@ napp.d by
!!i!8
thc 0l0E4V0l0E5
sden
sdd6 wrc foud
13.7 %
thu irdi@ting th. hislt prcv.bft. ofpenial
io hav. g/8r dclctions
vhil€
only 6.3 % of the dico&Ltionr
Miltal
et al.
(2004). c./s! d.laioD!
e
hd b*n
kroM
ro
s
B de Epon d G
[email protected]. rtoL of
&Dni.s
i!
this
egid
pridq tet. In tbit stldy
i! Patilt !i
Dopularioro tbrr
AZFC d€l€iiotu
ij
(Iable 4.3.1).
Eponcd in Nodh.n Iodia by
.xist in ih. haplogoup N
4
!
n
T.bL tt3.1:
Subj.cB wid o! wirbou
d.hlio!, i! dift@r b+loStoup!
13
Itg 43.1.
2%
Fig 4J.2: 29o
Aga]k s.l lbwin8
ihc
Agesc e.l ihlqilg th.
a'
6dr
! of Mulridq rcR
ofMulripld
PCR
forAab
fd AZFc
74
wiel dd n!9bgMp6 N3 .id N2 @ cun nUy wid.rpF.d i! $nc
@unil
Fpulrtid ofEat h EwF @GEto
hom
to
blE grltr &ldiola
rib.r.! !@pL i! Alia tr'I
lryBtcd
ar. (2003)
i!J!r.r
Th!
not
,r ot, 2007). Atuth6 h.plogloup D2b i3
chbaMba
D2b
N coolo! in J.!o and
i! EeFe rld ADdi@ popubti@, Rlppirg a
F6.E oflb! blplogrcup i3 BporBibl.
Epr*
niv.
e!d!
for
Edrrd fcnitity
ofgr/gr d.ldio@ @ lhoM in Fig 4.3-3..
€3 f,.plorFlDL!
tn
dd.'
ro
6!d
out
brplog$p.ictuion ein p.nid &lcri@ i! i.fcnilc
!!it r3 lidql io T.!L 3.3 rw s.d b! !.rHirg. El4.n
li!..98 (brplogreuF) @ &fd .!d tbir cLlio8bip, u!d.r ti. .lsuFion of
t!. Dilidu lEba of Durli@l ddt!, g@Et6 . re (Fig 3.2). IX.
!m@ld'G ed.r d@ by Y chl@o!@. co@ni@ (Ycc, 2002). Th.
brplosow !tudi6 tlw 3bu rh tlE 60.r t qud b+logow fo@d i5
go'+ r
d
hld%rcW
br YCC
of bid!.lic
3
eitb 52.9
i5 h.s.d
o
%
tb.
(I$lc
m
4.3.2). Th. hlplogro"p 3
Eor&lt Euirid
idcntifcd by tN. MU dclcti@ (Mcbdi .,
b.
tb.
tlqMt i! .ll P*isrui
s.
hlploeroup
al,
n6
1999).
aot d
i!
c+A of
cd
s Rhl
5RYl532
.rd
is
'nis d.l.rion ha .le fould ro
poF!!ri@ cxc.pr Hlritl
A.q@r h8 be!
e!, d.sirn
(Q!u
Nonh
.r al, 2002). Futth.r
Ernm ENF.nd Asi.
(zdjal a 4r, 199). Bishdl t!qu.!.y of 0ri! b+lo8r.!p i! folrd in th. greup
h.ving 3F@logcd.
mst
i! thi! h.plogoup.
a hlplo8tlup
But
This Esy
3
irdic.t to*td!
tlE hiSh
tlqu.rdy o.cu
in th.
Ft
of
P.tir..j
irf.rtility
75
{33. 2,,l Asnw sd dowils |b eDdncdi6 of 010E4!l(ts5. TL
@pk3 eithoin 6d'c.djioo
r!
lhorioS Dicro&Hi@ i!
|b
A.zFc
a
&
5
e
I
&
6
.c
77
lolulatiotr. Th@foE
i1
rculd
be
hisLlding to conclude rhnt th€ high
of irf.nility in the haplo8toup 3, H.plogrcup 3 b
*iib
.z@pc@ia
equal
Ih.
'1G58
% in
pBddt
d 'x'
j! e@!d'@ to tE
the haplosrcup
haploSmup
is
172, id€dtified by
t
Aon
ha
.160
diftMl .lhric
L &dlluc for Pujabi polulitioi foi
6nd our rhe
sislitr@r
's@iation
'ftid
sr.t
ftqllmy of ihi!
b..!
r.ported itr the
@Fdso8
i!
nlpbgrcup
5l fall in
Ml24
rotal
Psi ed
High
Sildhi
spLs i! ou id.nil.
Th@foc it is
@*!ry
Pujabi popdation
to
b &eiri!
nost Fvelant biplogroup is 2, which b d.fln d by having &cestal
bapLSrcup 2 which
m*6
&d lhis Fsntage is clo*
th4t is 7.3 % in
Ou.htr
7.E%. whn€
h P.ri$!d popuLti@
diffcEne. Sinildly M124 sle h!rc fou
t!.
Fpulrrio. wjrh
iof.nilig.
of hallosrclp 172 wilh
cpodcd 9.7 % oftLir haplogtup
of
l2f2
grups ofPujab wtiL m blplogml4 dal.
(0) or M9 a.d dqiv.d rrar. (l ) for SRY | 532.
our of
d.
ud obtrutivc eospmi. grclp.
pop'lali@ (FeDal co@uicltion). M6t of th.
Fpd.lic qw
P.kislrni popul.rioi
by YCC) hrvhS 15.6 70 in bfenne
high ftequ€ncy in olig6Feic
ieid.trc. of
in ob€tr@tiw
v&iou .r,h!i. 8rcuF (Qu.r 4 zr, 2002)
scond non
rLLlion (d*icl)aied
qEmy
A.qEBy. Abou 50 % of th. i.GltiL popdltion
b.lolgcd to this balloge'p, eiich b
v&yilg too
.ls prw€la
t
lakistni etbric
to
ou1
of thc fifty
i!. F.Dtly
gloups
showed
fou smpl6
Qdd
er al. (2002)
shosing no
si8lifi@r
oft @pl€s iLll n
caiculd€d
7.8%
pEulena of th.
(F6otul comuication). Othd
7E
baplogolts wbjch de r€po.t
fFqult
d io bc thc
in
Pakti.! @ l, l0 sd
28 but
ou id.niL popubtion c&i6 otrly on @plc &oo @l h.plogrcup, Tt af@
il
he
is o€d.d to
cl@
i!ftld.
Orc
lr4e
lebd
of
Esi@ a lhom in T.bL
AzF!,
ed c c8i@ hde ben Eloll.d
this srdy rh.r.
4.3.3.
d.letiN 9w lidcd
nal6. l.g. ,r al.
(
1999) poinr€d out
iEluding 19 L$4.iptio.al
uit
6!6t
ceion .ftcls olty orc
by El
Aqady.t.l.
tl.
(200r).
&lcti@
i!
Eowva
i!
sllnE in thc ft}ltid
dd€tion of tte wholc 3.5 M! egio(r
go. fliili6 of AAc
of bd. difr@ldrli@ rtat ir {[email protected].@is
rbc
F6o!.ppea
ro D. h*tthy.
Sinilr
!l@ obw.d in lhis 3ndy. IlEe mples hld conpl.le
a3.r Scc.Ls
of CDYrr/b
est
with
polForphius sdc s . tool
onsiB, which
d.pe
up@ sinplifi.d
polForphilbe @ s.ldtiv€ly
f.rrility
a
singl. neELorid.
CDY la
wimt
in this srtdy
AZF€
rctml app€M..
povDoiptliD
Y chl@osoad
w6 $Ee*d
d!!e a
sreup to
havins l|.sq
cooptisilg E distiEl
delelid esultiry in spmatoepnic
ro aflccr
c.g
Sioild
ro otrly thc sLon
th.t
Aprn nob th. [email protected] &il@,
quls EF
a.h eihlic
lhou th. Llg. d.letio! sFding AZFq
AzO053 ha
b ald c
b
AoD
asidior
aboln rhc hrplogorp
ofou s@pls
lep|6
for ihe coNiruction of phylogc!.ric
$suption
rairrl. W . sotu
polynortbis (M!!bd
5
C,/A
sin{Icd 7750
dd
not€d thai cDY la is
bp ar
Y
tbnr
all Y chrcnosotul
polrrorpbi!$ @ kloE
€r 44, 2004).
eDd of
ldl
asiai.d
In cDYl gee a
ion sirn.o<to!
wirh ioLniliry.
n
T.bb433: Stosiis iE
El'rbrD
R{t
ofd.Lii@ ioAA!, b.!dc r.8im.
arrt
AZf.
trot
2
DEY
.y\27
AZo{t1
A20 048
d.l
AZO 053
.Y!34
An.
DYst5{
DYS255
Dl\Z
lo
l3lra
of
lli! .lbb did3
in b@ozy8ou forn
ir i!ftrlilc gdp lh.
r.F...dirc
|ll rloM i! Fig 4.3.5. ft !s!.ho obw.d tbd, rh. 66 % of rh. rlllpLt
te bolt gr/gr &Llided CDYrt potldrtio. M..h.v.td. (200a) Epolt d
r! utr
thrl th. CDyla
tt
Pr!..d sny
iLLLd DAZ
poltldphs .lolg
!.1 of
gd.
Dair
Th.EfoE EoE
.!d DAZ copy [email protected]
DAZ3/4
of
d.L(@
'dDd
EGd
to
Fur6€r
s
wiih DAZ 3/ 4
F!
*o*
.N.
iddrility.
[d
@btc b dindDgrilh th. @py !@bd of
br*.u
thlt ho{
&leiid
@
ldy
is rcquiEd to
&d.d
of
N
to
Dlp th. &Ltiotr
incnd
@plc c.ny DAZI/2 &d
ud.atnd
1n.
ooplet .p.ctu
dd F6ohgb v|riniB in Y chlooo|oorl .tt.nc stiob wil
nldDlt ly lc.d to frdiDg
rt loluid
ofi!&ctiliry.
ll
ng a34! PWtr diSd er CDYI. dd
doeiq
F@
polyEo(Pbi6!,
ofbot CDYlr.!d b.
b
3
b.lrlr i!
spL
E2
SIJMMARY
Y
ctloroeo.
. uiqu ed lorcrtn slsrd
provid.s
hrploid sl!t@. Th. Y chsnolo4.
tugiig E@ uiqc
.6t
ro
drdy
&cudar.r all diff.Eir
th€
mullrioB in s
kinds of nutation
Dolrrdphi3d to hidly usrlbl. ni.jrcr.llii.s, Tbcs
mutitio., polrnorphi@ @ ncbin in d.t @i!i!g rb. Dopd.tid st&$e ard
oiSralory D.tleDs
*E
ed fo@ic inrtrsis. S.qMcc
difl@
of
$ncn d ton ihe ctnoE <btrblse ilfonatior,
F
Y sFcinc cbncs
hav. idetincd 6v.
rowl balclp.ci6c, polyboQhic bicbsar.llilcs. A sirglc oullipl.x ICR
s{s etup for 6.s.
lei
snd tl€n
polynoiplds
n.Ls 8on fouldn difld.nr Egio.ni
ion
rh. irdividu.l l@i @8.d
id.nti6.d.
70 out of
B{€@ tw
r
).
h.plotyp.s
\rft
ed
Pltistar IlF dit€sities of
154
ditrcF l hlplotyFs
p@nt i. o.ly
lrl
lh€ Mobdna 3howd vay
0,661 resp.cti*ly. Mltatio! ra|e
obs.ded ou1 of lhc 65
cu.nr
tool
i!
90
lo* divdity
wa ale
cultG on buarios .nq
30
for
ln.* ldi
.alculated in EBv
mutalims w@
gm6dod .rd tM
g.nq.tios Esulting in nuianon nrc of 0.3% IrEdoE
sct of well chi@tcued nicnsaielliie loci muld s.we .s
individual
Paldstan
l@i, tcw FPrn.doN id
rdrfoD.d cell lircs for th.e Dicrcs.lclliie Dek€6. Tm
Dlrniod lnd
(13 Evo) attd
An ov.6U haplogTe divmity of 0.961 in
fd t[.F
wE
singlc individt!&,
@I! fi.q@ntly, 9-8-17-1t}4
hicl| lomr of di*riDi!.tion
panicular. ihc KabEiri
lhat is 0,54
154
&d
@lat d
i!v.3t8Eted in ?56
populalions of
0.2209 to 0,793E
h.plo$Tes vcF fowd
9_a_r7_tr.?5 (E
swgcsd
th6.
s
lwlim
idlntii4lion od plFmiy
tcsling in
e
lhe
atLlitioml
fotsic m.dicile Rol€ of
8J
th. Y chrcn@6.
i!
E le iddtiliry h
wll 4lblish.d
s.d Y chbd@dd
a BFcibl. for ih. cdE d or @
speddoSd3is. AlEosl lGl5% cowle e hf.nilc @tDd th. Krtd &d in
d.l.ti6
hav.
apF.xi r.ly
hrlf of rh@ c.tar
crB
of
b.L
3lctuarog@sis. Ttc
bos.s
@6ida!d
difftnt.nviloMot
@y hflc
gcn tic
be!
&d
l or gcEtic
€us. Md n€qedy d@u4&d
inGnility is Y cEon@aal dclclioN
luu
c!@<tcs 80
devclopD.al
o.l. n lor b Bp@5iblq llGnilily h n8l6
ettltilg iop.ir.d
.mtlic apptinardv
Y cl!@()M.
FoLiG, $E. of qnich @ capoNibl.
60 M.gr
fd g.r
@ll
dd @intstl!@ ([email protected])- Y chrcn@dd gm's irvolvcd
i! sFnllocltFsis EGdt cid.
in AzFa. b dd c Egiod. USP9Y i!
a
ioporr&l
nmd@rt gcF in lh. AZFa Egioa RNA-bindils botif (RBM) fmilv in AzFb
rgio!
and SErcs
llerfoF
play
Fridine i! ihe
t cd.i6l ol.
in
AZFC rcgion
haL fenilit
hivc sFcinc .xPn$iod ir i'stis
l,
ln.
ptlgt
stdv w' cdi'd oul
del.lion u.lysis for AZF\ b md c Egions in 53 i'Jtnile individuab
hlv. shoM
2clo
del.timi in
A.zFa esion, 3 8vo
AzFc egion, wnich mish
Th. bain €ldidat
i! rl*
b.
FlpoDsibl€
'!d
lrr
Fsuls
deLlid3 in AZFb anc 5'8% in
fo! F te infeniliry in ?itistani
AzFc Fgron is lhc CDYI
td
DAZ
'lBt.r'
Multiplc
in AZF. Egid A
mpi.s of DAZ Cpn witl >996 of eqw@ idaritv
'xLt
potynoDhim in CDY1 (CDYI.) blve shoM.seiatin *ith bJenili9 13 5%
of this atlclc exists in hmozvgou fo@
i!
iDGnile grDup Psttid
d'laid
of AzFc
rdbd cd atso
Egion Gr/gr dclcliont Esdlhg in ihc itduciion of DAZ
'oDv
grcop
cds. infriility, V. hav. fodd 15% of thc grlgr d.l'lios in ou inf'fln'
t4
|'ryl. !da8 g/9 &Ld@ l! b6rl io !.s cDYlr
poltw!fir! d wll. hrtr vrqt ir d.d b trd.nhtl iL c.opLi.
od
66
|!.ctre
X ql 6.
of
uDd
vlll uldnr{|r L.d
@d
0o
ldolotic 1|.rdi@! h Y &mdold ft@ts i,tbh
fldilg lb rotrid br ifflily,
CE,{PIER 5
LITERATUR.E CIIED
Ahin-S.if6 L
E.,
t jdb., R L. Toui!.,
IL
nicrld.Ltim
Q., A.
rVF
ed*
M,@r, ra !&.il
Mrd|t,
2004. Y
.!lo'no.@.
saiDg b itftn c oc! in ttte: r [email protected]
F..ri.r h.!.d d $
Atr$
R lria.
of
ft@h
H@. R!!r.d. I9(4):?E&93
S.
s. R!}o|D, S. Siddiqi,
Kliiq, a- Mohtrddi4
M PtFiereq
A.
A-
lt o..4 r'
Picz2t,l L
qvdi-
Sftrrr, 3. Q. Mchdi. 2003. R@ostrdi@ of
h@r 4oltdoMv te
uiog loltDnPbic trdos.drl 6idsrr.Uitr.
Ao J !hl6
AIrb'oFl'
122(3):259{E.
Aylb
Q.,
a. Mobyuddi4 R Qea,
L Mt!.t,
T.
tld cld.iaixali@ of
TyLT-SDitt 2000. ld.tditrcri@
cbllmsdd oi@sLlitd A@ ,.qrncc
AcidlRe
c
Zaj4 s Q Mttdi
'!d
dda
dd tue
Y'
b.!' idoiDtlie Nu'leic
l52E(2):€E.
Baow,s.zluA. P&dt', T. zdj{
J-
xu Q sh!B-DqIl
Yan&
c Tvld'
d ib' ht|'@ Y
ch|t)o@o. Piicb FoviL! a etul rolvDdpbic 6and i! ChitF
Soith. 2000 MSY2:
.
slowlv aolving
popd.tiod. G*.244(l-2):29-33
EilieLUii'
86
Bd!e! A. V-, M. P..!,
P. T- Mchlosr! D,
c.lt@.
198. Evolqtio! ofRPS4Y.
Mol. Biol. Evol. l5(11):1412-9.
BeU, G. L lnd
I Jwb
1997.
ft.
Llsth distibutioo of r.rf.ct diod cp.driv.
DNA is
cGilErr *ih i& .volutio!
F@s-
J.
Mol. Evol. ,14:41+121.
1999-
Trld.n Ep..l fnda
Bdsn G.
N8ldc Acid
Bktdulpt
l. l9?.
Boeh
tld
E.
t
by
D
ubi.ld
silgb sLP dtn
Fogr@ io .!dvE DNA
do
Fq|l'B.
Rcs. 2?:573-s80.
Tribd of
Joblins; M.
Y .bb6oed.
qBRv3 @d
t.
A
4
$.
Kdb I!dr. hblicdiots' Kdicli'
nilttoo
2003. DuPliqtioB
ofth' AZFs ngi@ of ihc hud
D.diated bv h@ologou
coDpdible wilh dd€ f.nilitv.
r'@bilatiot
b'tw!
H@ Mold G'nd l2l
34t-347.
Boyd
S.
tt., A. F. s@lt,
.ll
Point
L lvt Ku*ct
and
nd.liolt *hic! @
teooslobi! @ino &rd
2(l):111.37-
.d
K D Soirh - l9E
@c..PtabL:
nelcotide
Tbe propdtion
s dliidc
lcqlatE C4,
J.
of
b's'd o!
Got! cttol'
E7
Btund€ll R. A,, A. Mielnil, D. Liotrq z. Ye, L, L, vc.ck, G. D. Pd€do, P. N.
scbleg.l. 1998- AzFb deletios pr.dict tbe
.b$e
t rticulq [email protected]&rid FElidinary cport of .
HE
R.prod
Miichcll, M. H. ,on6, T. B.
hllm
ih.
Cb,eicridid of th.
R-
prc8no61ic
e@it t6t
@di!s
P Edcles G LotrslPicd M.
atd N A
A6a
..!ll.@ !d fin. dPius
of lhc
H4Mvq
DFFRY g@ &d @nFdliv€
!r-b
sFd.lo@ qith
13: 2812-2815.
Brcm G. M., R A. FEI@g; C. A. s.4cnq
l99s.
of
H.
J
C@Le
dp6ior
aullsit &d dlppile lo
inrcdal oflh. noue Y chrcoo$me of
in Dfiv gce nM
Mols. G6d-, 7:9-107.
Chdg t. L., M. V, Sau.r dd S
fdtho
sd
his
DubEva dd
G
D.lisova.
ctrcoos.d' micodeletior in I
14: 2689'2694
M Woai.r, T
Gdttowski, I
I. Zrktuov. 2007. Y4brcmosoe hiplogoup N dbpesals
Sib€ria to
ENF J E@ C.ret
Didinget, D- aMl C. Schldt@ 2OO3
Mutadon
Y
fou irEnile sos. Hu. R.prod.
D.Frko M.. B. Maly@huk,
fiod souih
Bbm !99
?|!..s:
Te
Evid.!.c Frco
thG
52(9)1763-70
Dbrirct Mod's of MicblaLllit'
conpt't' Gdodic scqMes of
Nitu Sp@ic' Gemo. Rcsch.13:2242-2251.
8E
El
Asdy M.
K., S. F. El Sbltd, E- B!e.4 IC
M.giud. 2004.
Esptir Ml6
F.din A., A.
TB&i,
F.
Md*ul{
rtudy
d
L M. Shd..n md N. A.
Y chrcdoeF. uicrodcLtioB
eitb idioFrhic irfcnilit.
C@, E, Mdhin4
Ar.i
S.
Aiie
J.
Andol. M.i5(l):53-?.
Bdtati, A. C@tls, B, Eoel
FoEsr. 2005. ]\!s{.i!rid of Fnirr AZFC Egior &t
sp.66togdic [email protected] od hslc i.f€rtilily,
in
dd
lio6
C.
wnh
I &.d G.h.t. 42:209-
2t3.
F.rlin A-, E. Morc, A. C&olla C. Fodstr. 199.
IIllDs oaL tnf.fiitiry ad y
cbrcDoson {telaioB: ble of AZI cddidsr. 8€B DAZ, RBM ed
DFRY. Hu. R.!u$d. 14:l?10-1715.
F.din A., E- Morc. A. Ro$i, B. Dalapic&h ald C. Forsta. 200j. Ttc
cbll@tu t 84cpdnh 6dd
md ddction enlysis in infertiL
F.hd<les
S., S.
2003. A
6.n
Fitul
b (Azrb) a8ion: s€qucc.
nd.
J.
M.d.
C.ftr,
40(l
): I
hMe y
sflf,re,
&24.
Pe&chini, L. H. Meyd, C. Ibndia, C. Tyld-SDiiq
p.
}l. vogi
L!e. AZFo .Llerio! Edov6 DAZ3/DAZ4 ald lEby 8.16 fro6
in Y blplogrcup N, Am. J.
Hu,
S., S. Kbaliq, A. Mohyrddi4 M.
Gncr.
74: I
E0-7.
pa!.iomu, C. T. SDirL p.
Unddhill, Q. Atrb, 200?. Y ctuonosoodt evidc!@ fo!
i
A.
lioited CF.k
E9
cffiihrd@
F@r.
b
rh. Prthro lopulalid of P.Li$!e Euopa! Jouml of
h@dg.&ric!.
15,
12l-126,
C., A.
Fqli! a/l
E.
MN.
g.@
on th€
ir'fer
i,V.
2000.
D.t
tid .rd .,Q!si@ ,!rlFj!
hu!@ Y chDno$a. Evet
HM.
Motec.
da
mjo.
FIG for DBy
hn
L
c.reL' 9t tr6r- 1\69.
FoBtr, C., A, C@U L. Berol@i. A. B.h.U4 A.
.homalitiq doas &vdcly oligGlemic
ifi&ytopLsic rF@ inj..tio!.
Pu Y.X,
of A-zFt
J.
nd
Fcitin-2o0j_ CEnaic
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Cli!. Endoq_ M.rab.
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C.
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Sibly
R I. Kotosta M, Dobi8tr
Mictu$t
llit
niwtah
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R
/acaA
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od
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P.
h
LiA
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Ma[oFini@,
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in
G.
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E.
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of
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Tylc.SritL
C,
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A. Mohylddia s. F!, P. Li, N. Yllllrstcv., R.
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Ao.
Zbu Y., D. C. Q@[d
ad
J.
and
Hu.
J. E.
Ruib.bev,.t
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c. TtlcFsoirL
Qdd, Q Ayub,
I
Xu, Q- shq
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2003. The seenc
B.
t8e.y of
G€@t. 72(3):7u-21.
Str6tu.
2000-
b*rtios, sulttituiios $d rh.
disin ofnicrcsat ll:n6. G@r, R.s.76(3):227-X6.
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shon @mruication
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