The Ocurrence of Multi-Nucleated Cells in Parathyroid Glands of

Y.Y,Ü. Vet. Fak. Derıı , 1996. 7(1 -2): 54-57
The Ocurrence of Multi-Nucleated Cells in Parathyroid
Glands of Sheep and Rats
Mehmet KANTER
Yüzüncü YıL Üniversitesi. V~teriner F~kaltesi. Hisloloji ve Embıiy(lloii Anabilim Da lı . Van. TÜRKIYE
Geliş tarihi: 14 Haı:iran 1996
Koyun ve
Sıçanlarm
Paratlroid Bezlerinde Multl-Nilkleer Hücrelerin
Oluşumu
Sum mary: The effe cts ol different f<Xation m ethods on the uıtrastructure of paral hyroid eclis was st udicd in sheep and ıats. Electron
mıcroswpy of pa ıa lhyıoid (PT) glands ol sheep fixed by immerslon demo n stıated mutıi-nucfeated cells wilh ctose lo each otheı
irregu laı ly shaped nuciei showing fOklS and undulations. wrth remnants of the plasm a membıane between them. The chro matın Ls fi nely
granular and dispersed over the nucleus in irregular clusterıı. The electron-l ight cytop lasmic matrix wnl ain in9 few secml ory 9ra nules
(SG), smail Golgi wmple" (Ge). rough endoplasmic rellculum (RER) with dilatcd cislemae were ind icated. Mitochondria were markedly
sW<lllen . In e<mtrast. the PT glands of the rats fixed by perfusion exhibited unilorm rells instead of muHi-nuclea r cells. The uniform cells
conta ining electron..ıense cytoplasmic matri.>;, toıluous ptasma membranes. slighlly dilaled RER ;;;ınd Ge, few SG and incanspıcuous
dilated mitochondria \Yere observed . From Ihese finding we eonelude that the oecurrence of the -multi-nucleated eells· in PT glands of
shccp dcpends on fixation methods.
Key Words: Parathyrokj gland,
mul!i-nude~ted
eeL! . !ixalion method
Özel: l:Iu çal ı şmada. farkl ı fiksasyon metoUanmn koyun ve ıat paratiroidle~nin uıtrastrOkHlrO Oıerine etkisi incelendi. Immcrsiyonfiksasyonla tespit edi~n koyun parellro<d (PT) bezleıinin eicktron mikroskopisinde, multi-nOkleer hüctelerde düzensiz şekilli
çekirdekicrin bir birine çok yakın oldu!!u ve aralarında plazma membran kalınt ılarını n bulundu!ju belirlendi. Kromı:ıtln'in , çekirdek
içinde ince Ilr~nül ler ve dOıensiz psrçacıklar şeklinde da!jıkl ı !j' görüklO. Y~unlu9u az alan srtoplazmik matrikste, az sayıda salgı
granülüne ve Golgi kompleksinin kOçOk kesedkıerine rasııandı. Endoplazmik retikulum sistemalar ı ve mrtokondriyonların oldukça fazla
şek i lde dilale oldu!!u tesbit edi ldi, Perfü~on-fiksasyonla lesp~ edifen ral PT bezlerinin elektron mikroskop incelenmesinde ise. multinlı klee' lrüCrt.!e1 yerine uniform hilereler gözlendi. Bu hOcrelerde plazma membranının kıvrımh ve sitoplazmik malriksin ~un okııığu
belirlendi. Endoplazmik retikulum sistema lan . Golgi kompleks kesecikteri ve m~okondriyonların önemsiz derecede dilate olduğu ve
salgı granüllerine çok az rastlandı!!ı tesbit edildi. Bu bulgulardan , koyun paratiroid bezlerinde multi-nükleer hücrelerin oluşumunun farklı
fii<sasyon metollarından kaynak l anabi l eceği sonucuna varıld ı ,
Anahtar Kelimeler: Paratiroicl bezi, multi-nUkleer hücre. fiksasyon metodu
Introduction
Histologically, the parathyroid (PT) cells
comprise dark chief cells, lighl ehief cells and oxyphil
cells (13). In addition to these cells, occasional
menlion is made in the literature of mu lti-nucleated
cells or syncytial cells (10). Theyare reported to
occur in parathyroid glands in man (1) and dogs (3,
6. 14, 17) regardless the age or funcUonal stage
(12). Multi-n ucleated ceIls are especially found in
hyper-paralhyroidism (14, 21).
The first eleclron-microscopical description
of parathyroid was by Ekholm (7). later, there have
been many eleclron-microscopical investigations on
the normal, slimulated and inaclivaled paratlıyroid
glands in different species; these have been well
documenled by Nilsson (13). At the eleetron
microscopic level. PT glands contained 5 cen
variants: dark cells, light cells, atrophic cells,
inlermediale cells and mulli-nucleated cells (22). In
dogs and rats, mu ı ti-nuclealed cells were only
obser,,' j in PT glands fixed by immersion in
glutaraldehyde (19) bul never in paralhyroid glands
after perfusion fixation (22).
In the present study, we first compare the
ultrastructure of PT glands of sheep fixed by
immersion with that of glands of rals fixed by
perfusion using Ule same fixative and second
evaluale the influence of different fixation melhods
on the ullrastructure of PT eeIJs. The aim of this
study was to investigate the occurrence of the multinucleated ce!!s in parathyroid glands of sheep and
rals whether depends on fixation methods.
Materials and Methods
Parathyroid
glands
for
electronmicroscopical
examination
were
obtained
immediateıy after 3 sheep were slaughtered. The
glands were immediately placed into ice cooled
phosphate buffered saline (PBS). dissected into
smail blocks and immersed in 2.5 % glutaraldehyde
(GA) in ü.1 M Na/K-phosphate for 2 h at 4 ·C under
gentte shaking. Three adult rats (200-300 g.) were
deeply aneslhetized by intraperitonaea! injection of
Ketalar (65 mg/kg). Parathyroid glands were
removed (under the aid of a stereomicroscope) after
perfusion with 2.5 % glutaraldehyde in 0.1 M. Na/K-
Y Y.O. Vet Fak. Derg. 1996. 7(1-2): 504-51
phosphate at room temparattıre for 15-20 min. with a
pressure of approximately 120 cm H20 through the
left cardiac ventncle. Then, Ihe glands were
dissecled and immersed into ice-cooled 2.5 %
glutaraldehyde in Ihe same buffer for 1 h undar
genUe shaking. Atler immersion-and peıfu5ion
fixations tissue blocks 'NEre Ihen washed in the
same bl1ffer for 1 h at 4 ·C or slored al 4 ·C
overnight, postfixed with 1 % osmium lelrexlde
(0504) in 0.05 M same buffer for 1 h et 4"C,
dehydrated in a graded series of ethanol starting et
70 % e&eh slep for 7 min. and atler two changes in
propylene oxide. The tissues were embedded in
Epon 812. Semithin seclions were stained with
methylene blue and ultrathin secbons with Mguranylecetate and lead citrate.
Results
Light micrescopic examination of methylene
section revealed a
blue stained semithin
heterogeneous appearance of PT ooUs in glands
fixed by immersion whereas in pertusion fixed PT
glands the parenehymal ooUs showed a high
unifonnity.
Electron microscopy of PT glands of sheep
fixed by immersion demonstrated multi·nuelealed
cells with clese lo each other Irregular\y shaped
nuelei showing folds and undulatians, with remnants
of the plasma membrane bemeen them. The
chromatin is finely granular and dispersed over the
nudeus in ifregular dusters. The eleeıron-lighl
cytoplasmlc matrix containlng none or only few
secrelOl'Y granules (SG), smail Golgi complex (GC),
reugh endoplasmlc reticuium (RER) wılh dileled
dslernee were indicated. Milochondria were
markedly swollen (Fig, 1). In conlresl, the PT glands
of rals fixed by perfusion did nol exhibit any multinucleatecl oolls. In eddition, perfusion fixed PT cells
were very uniform so that the distinction between
dark and light clıief eells were impossible. The
uniform cells conıaining eleclron-dense cytoplasmic
matrix. lortuous plesma membranes: slightıy dileled
RER and GC, few SG and inconspicuous dUaled
mitochondria were observed (Fig. 2).
Figuro 1.Sheep PT cells fixed by immersion wilh 2.5 % GA in 0,1 M NalK-phosphate for 2 h al 4 ·C and
postfixed with 1 % OS04 in 0.05 M Na/K-phosphale for 1 h al 4 ·C. Part of a multi-nueleated eells wilh ciose
lo each other irregularly shaped nuclei Showing folds and undulationS,with rernnants of the pJasma
membrane between !hem. The chfomatin is finely granular and dispersed over the nueleus in irregular
clusters. The eleclren-light cytoplasmic matrix conlaining none or only few secretory granules (SG), smail
Golgi complex (GC), fOugh endoplasmic relleulum (RER) with dilated cisternae and markedly swollen
milochondria (m). (EM x 7400).
55
Y.Y.Ü. Vet. Fak. Derg. 1996, 7(1 -2): 54-57
Figure 2. Rat PT cells fixed by perfusion with 2.5 % GA in 0.1 M Na/K-phosphate for 15-20 min. at room
temperature and postfixed with 1 % Os04 in 0.05 M Na/K-phosphate for 1 h at 4 ·C. Uniform cells containing
electron-dense cytoplasmic matrix, tortuous plasma membranes, slightly dilated RER and GC, few SG and
inconspicuous dilated mitochondria (m). (EM x 13000).
glands which were fıxed by perfusion under carefully
controlled conditions, multi-nucleated cells were
Multi-nucleated cells in parathyroids were never observed (18, 20, 23). Since this cell type
first described at the beginning of this century (10) . exhibit largely distended RER cisternae and
remnants of plasma membranes (3, 12), it is
Their abundant presence in the present material is
typical of parathyroid hyper-function as described by believed that multi-nucleated ce lls are the result of
Dammerich (6). It is understandable that the number disintegration of cell membranes taking place during
of multi-nucleated cells increases with age, as fıxation or subsequent dehydration and that theyare
not norrnal constituents of canine PT glands.
reported by Bargmann (1) and Setoguti (17).
Glutaraldehyde reacts rapid ly but penetrates
There has been much discussion about the
development of multi-n ucleated cells in animals. The . slowly (11) . FA, on the other hand , needs 24 h for
most common polykaryocytes are foreign body giant fixation but it has the advantage of rapid penetration
cells and osteoclasts, which develop by fu sion of (8). Therefore, immersion of precooled PT glands
several mononuclear cells (5, 15). The syncytic into a formaldehyde-glutaraldehyde mixture seems
trophoblasts in the placenta (4) are morphologically to reduce the formation of "multi-nucleated cells"
similar to the multi-nucleated cells in the parathyroid,
(19). However, this procedure does not prevent the
observed by both light and electron-microscope, The development of other fixation artefacts in dog and rat
syncytiotrophoblasts are presumed to be formed PT glands such as shrinkage of nuclei, dilation of
th roug h fusion of several trophoblasts (1 6, 9). RER cisternae, distribution of Golgi lamellae,
However, contrary opinions are also found in the swelling of mitochondria and whole cells.
literature: it is suggested that theyare a resu lt of
At present it is not clear how multi-nucleated
cells arise. However, the undulated nuclei, the
nuclear division of one cell (2). Oksanen (14)
suggested that mitoses are not seen in the multi- swollen mitochondria, the dilated RER cisternae and
nucleated ce lls or in the chief cells. The multi- the remnants of plasma membranes indicate that
nucleated cells are considered to be living and
"multi-nucleated cells" are the result of a rapid
secreting ce lls and not degenerating chief cells (6).
degenerative process. This process can take place
They were regularly see n in hyper-functioning within minutes before or during immersion in
parathyroids (14).
glutaraldehyde. From these finding we conclude that
In morphometric studies on the behaviour of the occurrence of the "multi-nucleated cells" in PT
cell membranes involving more than 200 rat PT glands of sheep depends on fıxation methods.
Discussion
56
y.v.O. Vet. Fak. Der;. 1996, 7(1-2): 54-57
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