Ratlarda beyincik korteksinde yaşa bağlı değişimlerin histolojik ve histometrik yöntemlerle belirlenmesi
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Dosyalar
Tarih
2024
Yazarlar
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Yayıncı
Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, geçmişten günümüze kadar gerek bilim adamları gerekse tüm insanlar tarafından merak konusu olan normal yaşlanma sürecinin beyincik dokusu üzerindeki etkilerinin histolojik ve histometrik yöntemlerle araştırılması amaçlanmıştır. Bu amaçla 24 adet Wistar albino ırkı erkek ratlar kullanıldı. Ratlar genç (4-6 haftalık, n:8), yetişkin (20-22 haftalık, n:8) ve yaşlı (22-24 aylık, n:8) olmak üzere 3 gruba ayrıldı. Anestezi altında servikal dislokasyon ile sakrifiye edilen ratlardan beyincik dokusu örnekleri alındı ve %10' luk formol solüsyonunda tespit edildi. Tespit edilen dokular rutin histolojik yöntemlerle takip edilerek parafinde bloklandı. Bloklardan 5-6 μm kalınlığında seri kesitler alındı. Seri kesitlerde genel histolojik değerlendirmeler ve ölçümleri gerçekleştirebilmek için Hematoksilen-Eozin, Kluver-Barrera ve PAS reaksiyonu ile astrositlerdeki GFAP ekspresyonunun belirlenmesi amacıyla GFAP immünohistokimyasal boyamaları yapıldı. Hazırlanan preparatlar ışık mikroskobu altında incelenerek beyincik dokusunda histometrik ölçümler yapıldı ve bu ölçümler kaydedildi. Tüm yaş gruplarında folyumların tepe noktalarında yapılan histometrik ölçümler sonucunda moleküler katman kalınlığının genç ve yaşlı gruplarına göre yetişkin grubunda belirgin bir şekilde arttığı, granüler katman kalınlığının ise genç rat grubuna kıyasla yetişkin ve yaşlı gruplarında belirgin bir şekilde artış gösterdiği tespit edildi (P<0,05). Toplam korteks kalınlığının tüm yaş grupları arasında istatistiksel olarak anlamlı farklılıklar gösterdiği ve en kalın korteks katmanının yetişkin grubunda olduğu dikkati çekti (P<0,05). Pürkinje hücre sayısının yetişkin grubuna göre yaşlı grubunda belirgin bir şekilde azaldığı ortaya kondu (P<0,05). Pürkinje hücre sayılarındaki azalmanın yanı sıra 22-24 aylık yaşlı ratlarda Pürkinje hücre sitoplazmalarında PAS pozitif yaşlanma granülleri tespit edildi. İmmunohistokimyasal değerlendirmeler sonucunda folyum bazında farklılıkların olmasıyla birlikte genel olarak yaşlı ratlardaki GFAP-immunoreaktif astrositlerin genç ve yetişkinlere göre önemli ölçüde arttığı, bu duruma astrositlerin hücre gövdelerinin belirgin bir şekilde fark edilen hipertrofisi ve güçlü GFAP immünoreaktivitesinin eşlik ettiği ortaya kondu. Yaptığımız bu çalışmada uzunluk ve genişlik bağlamında yaşa bağlı folyum bazında farklılıkların olmasıyla birlikte genel olarak genç gruptan yetişkin gruba doğru uzunluk ve genişlik ortalamalarının artış gösterdiği, yaşlı grupta ise yetişkin grubuna göre uzunluk ve genişlik ortalamalarının azalma eğiliminde olduğu tespit edildi. Sonuç olarak yaşlanmanın beyincik üzerinde etkili olduğu sonucuna varıldı.
The aim of this study was to investigate the effects of the normal aging process on cerebellar tissue, which has always aroused the curiosity of scientists and all people, using histological and histometric methods. For this purpose, 24 male Wistar albino rats were used. Rats were divided into 3 groups: young (4-6 weeks old, n:8), adult (20-22 weeks old, n:8) and old (22-24 months old, n:8). Cerebellar tissue samples were taken from rats sacrificed by cervical dislocation under anesthesia and fixed in 10% formalin solution. The tissue samples were processed by routine histological methods and blocked in paraffin. Serial sections of 5-6 μm thickness were taken from the blocks. Hematoxylin-Eosin, Kluver-Barrera staining, and PAS reaction were carried out to perform general histological evaluations and measurements on serial sections, and GFAP immunohistochemical staining was performed to determine GFAP expression in astrocytes. The slides were examined under a light microscope and histometric measurements were made in the cerebellum tissue and these measurements were recorded. As a result of histometric measurements performed at the tops of the folia in all age groups, it was determined that the molecular layer thickness significantly increased in the adult group compared to the young and elderly groups, and the granular layer thickness significantly increased in the adult and elderly groups compared to the young rat group (P<0,05). It was noted that the total cortex thickness showed statistically significant differences between all age groups and the most thickest cortex layer was found in the adult group (P<0,05). The highest mean Purkinje cell linear density was found in the adult group (P<0,05). In addition to decreasing of Purkinje cell linear density, PAS positive aging pigment granules were demonstrated in Purkinje cell cytoplasm in 22-24 months old rat groups. As a result of immunohistochemical evaluations, it was revealed that, although there were differences on the basis of folia, GFAP-immunoreactive astrocytes in old rats were generally significantly increased compared to young and adult rats, and this was accompanied by a clearly noticeable hypertrophy of the cell bodies of astrocytes and an strong immunoreactivity of GFAP. It was determined that although there were age-related differences in the average length and width generally increased from the young group to the adult group, and that the average length and width tended to decrease in the older group compared to the adult group. In conclusion, it was demonstrated that aging has an effect on the cerebellum.
The aim of this study was to investigate the effects of the normal aging process on cerebellar tissue, which has always aroused the curiosity of scientists and all people, using histological and histometric methods. For this purpose, 24 male Wistar albino rats were used. Rats were divided into 3 groups: young (4-6 weeks old, n:8), adult (20-22 weeks old, n:8) and old (22-24 months old, n:8). Cerebellar tissue samples were taken from rats sacrificed by cervical dislocation under anesthesia and fixed in 10% formalin solution. The tissue samples were processed by routine histological methods and blocked in paraffin. Serial sections of 5-6 μm thickness were taken from the blocks. Hematoxylin-Eosin, Kluver-Barrera staining, and PAS reaction were carried out to perform general histological evaluations and measurements on serial sections, and GFAP immunohistochemical staining was performed to determine GFAP expression in astrocytes. The slides were examined under a light microscope and histometric measurements were made in the cerebellum tissue and these measurements were recorded. As a result of histometric measurements performed at the tops of the folia in all age groups, it was determined that the molecular layer thickness significantly increased in the adult group compared to the young and elderly groups, and the granular layer thickness significantly increased in the adult and elderly groups compared to the young rat group (P<0,05). It was noted that the total cortex thickness showed statistically significant differences between all age groups and the most thickest cortex layer was found in the adult group (P<0,05). The highest mean Purkinje cell linear density was found in the adult group (P<0,05). In addition to decreasing of Purkinje cell linear density, PAS positive aging pigment granules were demonstrated in Purkinje cell cytoplasm in 22-24 months old rat groups. As a result of immunohistochemical evaluations, it was revealed that, although there were differences on the basis of folia, GFAP-immunoreactive astrocytes in old rats were generally significantly increased compared to young and adult rats, and this was accompanied by a clearly noticeable hypertrophy of the cell bodies of astrocytes and an strong immunoreactivity of GFAP. It was determined that although there were age-related differences in the average length and width generally increased from the young group to the adult group, and that the average length and width tended to decrease in the older group compared to the adult group. In conclusion, it was demonstrated that aging has an effect on the cerebellum.
Açıklama
Anahtar Kelimeler
Beyincik, GFAP, İmmunohistokimya, Yaşlanma, Rat, Cerebellum, Immunohistochemistry, Aging
Kaynak
WoS Q Değeri
Scopus Q Değeri
Cilt
Sayı
Künye
Kuşat, T. (2024). Ratlarda beyincik korteksinde yaşa bağlı değişimlerin histolojik ve histometrik yöntemlerle belirlenmesi. (Doktora Tezi). Selçuk Üniversitesi, Sağlık Bilimleri Enstitüsü, Konya.
