Maternal retinol asetat (vitamin A) uygulanımı takiben yenidoğan rat hipokampus Ca1 bölgesinin ışık, elektron, immunohistokimyasal ve morfometrik yöntemlerle incelenmesi
Küçük Resim Yok
Tarih
2001
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/closedAccess
Özet
ÖZET Çalışmamızda ; maternal olarak uygulanan Retinol Asetat (VitA)'m loza bağımlı teratojenik etkisinin, santral sinir sistemi ve özellikli olarak ripokampus dokusunda oluşturduğu ultrastrüktürel değişikliklerin incelenmesi maç edinilmiştir. Maternal olarak tek doz 40000Ü/kg Retinol Asetat (VitA), 160000Ü/kg tetinol Asetat (VitA) ve %0.9 serum fizyolojik (kontrol grubu) uygulanımını akiben gebelik sonlanana dek beklenip, postnatal l.gün intrakardiak perfüzyon işleminden sonra yenidoğan sıçanların hipokampal dokuları çıkarılıp ışık elektron mikroskobik, immünohistokimyasal rutin takipleri yapıldı, parçalar ışık re transmisyon elektron mikroskobu (TEM) ile incelendi. İmmünohistokimyasal çalışmamız için vimentin ve nöron spesifik enolaz adlı belirleyiciler ile çalışıldı. Kontrol grubunda belirgin hücre proliferasyonu, ökromatik nöronal nükleus, belirgin ve düzgün yerleşimli nükleoluslar, düzenli hücre membranı, düzenli nükleer mebran yapısı, sağlam mitokondrial yapılar, akson ve dendrite ut transvers kesitler izlenirken kontrol grubundan anlamlı derecede farklı olarak Retinol Asetat uygulanan gruplarda doz artışına paralel biçimde artan nörotoksisite bulguları saptandı. Bu bulgular nörotubulus ve nörofilamentlerde dezorganize akümülasyon, piknotik nükleus, sitoplazmik protrüzyon,, interstisyel, ntrasitoplazmik ve intraaksonal ödem, nükleer membranda ilerlemiş tonfigürasyon düzensizliği, genişlemiş granüllü endoplazmik retikulum yapıları, aksoaksonal ve aksodendritik sinaps, presinaptik membrana sahip aksonda yoğun veziküler yapılar (özellikle 160000Ü/kg retinol asetat uygulanan grupta), mitokondri ve krista yapılarında şişme ve lizis, fıbröz astrositer uzantılar, ribozomal dağılma, kısa dallantılı dendrit, belirgin intersellüler ayrışma (özellikle 160000Ü/kg retinol asetat uygulanan grupta) olarak izlendi. İmmünohistokimyasal incelemedeki sonuçlar ışık ve elektron mikroskobik sonuçları destekler nitelikte idi. Nöron spesifik enolaz adlı belirleyici ile nöronal heterojenite, akson ve dendrit hasarlanması kontrol grubuna kıyasla oldukça belirgindi, hücre içi iskelet proteini olan vimentin ile yapılan çalışmada kontrol grubundan farklı olarak ödem ve hasarlı alan çevresi dezorganize radial tarzda tutulum belirgindi. Tüm bunlara ilave olarak ödemli hücre ve normal konfigürasyondaki hücreler sayıldı ve morfometrik analizi istatistiksel olarak yapıldı. Kontrol grubu ile retinol asetat verilen gruplar arasında anlamlı düzeydestatistiksel farklılıklara rastlandı. Ödemli hücre grupları sayısal analizinde en çarpıcı sonuçlar son grupta yani 160000IU/kg retinol asetat uygulanan grupta bulundu, toplam sayılan 924 hücreden 503 tanesinde idem bulgusuna rastlandı bu % 54.43 gibi bir orana denk geliyordu ve doz artışına paralel olarak hücre hasarlanması ve ödem artışını kantitatif olarak belirtiyordu. Kontrol grubundaki % 8.79'luk oran ve bu oranın yüksek doz retinol asetat uygulanımı ile % 54.43 'e çıkması retinol asetat ve teratojenite anlamında önemliydi. Gruplar arasında istatistiksel olarak anlamlı farklılık olup olmadığı incelendiğinde; kontrol grubu ve 40000 IU/kg retinol asetat uygulanan grup ırasında anlamlı (*),kontrol grubu ve 160000 IU/kg retinol asetat uygulanan grup arasında oldukça anlamlı (***) ve retinol asetat uygulanan gruplar arasında anlamlı (*) istatistiksel farklılık saptandı
SUMMARY In our study it was aimed to study the ultrastructural variatons caused by dose dependant teratogenic effect of maternally applied retinol acetate (Vit A) on central nerve system and especially on tissue of the hippocampus. After maternally application of single doses of 40000 IU/kg retinol acetate, 160000 IU/kg retinol acetate and %0.9 serum physiologic (control group) waited until the pregnancy was over. After the postnatal first day intracardiac perfusion process, the hippocampal CA1 tissues of newborn rats dissected and after their routine immunohistochemical follow-ups under a light microscope were done, the parts were examined by a light and transmission electron microscope ( TEM). For our immunohistochemical study it was worked with determinants called vimentin and neuron specific enolase. The following findings detected in control group; evident cell proliferation, euchromatic neuronal nucleus, nucleolus with evident and well arranged settlement, systematic cell membrane, structure of systematic nuclear nembrane, sound mitochondrial structure. In addition, while transverse cross- sections belong to axon and dendrite were being reviewed, in the groups retinol icetate was applied an increasing neurotoxicity in parallel with dose elavation was found, which was significantly different from control group. The following findings were detected;disorganized accumulation in leurotubulus and neurofilaments, pyknotic nucleus, cytoplasmic protrusion, interstitial, intracytoplasmic and intraaxonal edema, advanced configuration irregularity in nuclear membrane, endoplasmic reticulum structures with enlarged granules, axoaxonal and axodendritic synapse, utensive vesicular structures in the axon that has presynaptic membrane [specially in the group 160000 IU/kg retinol acetate was applied), swelling ind lysis in mitochondria and crista, fibrous astrocyter extansions, -ibosomal distribution, short branched dendrit, apparent intercelluler lissociarion (specially in the group 160000 IU/kg retinol acetate was ipp!ied).The results of the immunohistochemical research have supportive characteristic against results obtained from light and electron microscope. rhe determinants called neuron specific enolase and neuronal heterogenity, axon ind dendrit damaging were considerably more obvious comparing to control şroup. In the study performed with vimentin that was intracell protein, in the >eriphery of the edema and damaged area different from in the control group liffusion in a way of disorganized radial was obvious.In addition to these the cells with edema and the cells within the normal configuration were counted and the morphometric analysis statistically achieved. Statistically significantly differences were encountered between control group and retinol acetate given groups.In the quantitative analysis of cell groups with edema the most impressive results were obtained in the last group, that is, the group 160000 IU/kg retinol acetate was applied. From totally counted 924 cells edema detected in 503, this was equal to a rate of % 54.43 and it was detennining the elevation of edema and the cell damage in parallel with dose increase. In control group the rate of %8.79 and the increase of this rate to % 54.43 by application of high dose retinol acetate was important from the view of retinol acetate and teratogenity. When it was studied to find out whether there was a statistically significant difference between groups, a statistically significant difference was detected between control group and the group 40000IU/kg retinol acetate was applied (*), a highly statistically significant difference was found between countrol group and the group 160000 IU/kg retinol acetate was applied (***), and there was a statistically significant difference between control group and the group 40000 IU/kg retinol acetate was applied (*).
SUMMARY In our study it was aimed to study the ultrastructural variatons caused by dose dependant teratogenic effect of maternally applied retinol acetate (Vit A) on central nerve system and especially on tissue of the hippocampus. After maternally application of single doses of 40000 IU/kg retinol acetate, 160000 IU/kg retinol acetate and %0.9 serum physiologic (control group) waited until the pregnancy was over. After the postnatal first day intracardiac perfusion process, the hippocampal CA1 tissues of newborn rats dissected and after their routine immunohistochemical follow-ups under a light microscope were done, the parts were examined by a light and transmission electron microscope ( TEM). For our immunohistochemical study it was worked with determinants called vimentin and neuron specific enolase. The following findings detected in control group; evident cell proliferation, euchromatic neuronal nucleus, nucleolus with evident and well arranged settlement, systematic cell membrane, structure of systematic nuclear nembrane, sound mitochondrial structure. In addition, while transverse cross- sections belong to axon and dendrite were being reviewed, in the groups retinol icetate was applied an increasing neurotoxicity in parallel with dose elavation was found, which was significantly different from control group. The following findings were detected;disorganized accumulation in leurotubulus and neurofilaments, pyknotic nucleus, cytoplasmic protrusion, interstitial, intracytoplasmic and intraaxonal edema, advanced configuration irregularity in nuclear membrane, endoplasmic reticulum structures with enlarged granules, axoaxonal and axodendritic synapse, utensive vesicular structures in the axon that has presynaptic membrane [specially in the group 160000 IU/kg retinol acetate was applied), swelling ind lysis in mitochondria and crista, fibrous astrocyter extansions, -ibosomal distribution, short branched dendrit, apparent intercelluler lissociarion (specially in the group 160000 IU/kg retinol acetate was ipp!ied).The results of the immunohistochemical research have supportive characteristic against results obtained from light and electron microscope. rhe determinants called neuron specific enolase and neuronal heterogenity, axon ind dendrit damaging were considerably more obvious comparing to control şroup. In the study performed with vimentin that was intracell protein, in the >eriphery of the edema and damaged area different from in the control group liffusion in a way of disorganized radial was obvious.In addition to these the cells with edema and the cells within the normal configuration were counted and the morphometric analysis statistically achieved. Statistically significantly differences were encountered between control group and retinol acetate given groups.In the quantitative analysis of cell groups with edema the most impressive results were obtained in the last group, that is, the group 160000 IU/kg retinol acetate was applied. From totally counted 924 cells edema detected in 503, this was equal to a rate of % 54.43 and it was detennining the elevation of edema and the cell damage in parallel with dose increase. In control group the rate of %8.79 and the increase of this rate to % 54.43 by application of high dose retinol acetate was important from the view of retinol acetate and teratogenity. When it was studied to find out whether there was a statistically significant difference between groups, a statistically significant difference was detected between control group and the group 40000IU/kg retinol acetate was applied (*), a highly statistically significant difference was found between countrol group and the group 160000 IU/kg retinol acetate was applied (***), and there was a statistically significant difference between control group and the group 40000 IU/kg retinol acetate was applied (*).
Açıklama
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Anahtar Kelimeler
Morfoloji, Morphology