Gelişimsel olarak immatür gonad hücrelerinde oksitosin reseptörünün incelenmesi
Developmental investigation of oxytocin receptors in immature gonad cells
Yazar: FIDAN PASHAYEVA
Danışman: PROF. DR. GÖZDE ERKANLI ŞENTÜRK
İstanbul Üniversitesi-Cerrahpaşa / Lisansüstü Eğitim Enstitüsü / Histoloji ve Embriyoloji Ana Bilim Dalı
Derece: Yüksek Lisans, 2023
Konu: Histoloji ve Embriyoloji = Histology and Embryology
Dizin: Over = Ovary ; Testis = Testis

Özet:
Oksitosin reseptörü, birçok dokunun yanı sıra over ve testis dokusunun çeşitli hücrelerinde ekspresyon gösteren bir nöropeptid hormon olan oksitosinin reseptörüdür.  Gelişimsel olarak dişi ve erkek gonadlarında oksitosin reseptörünün dağılımı ile ilgili kısıtlı bilgi bulunmaktadır. Bu çalışmada, fare üreme sistemi gelişiminde immatür gonad hücrelerinin OTR ekspresyonlarının farklı periyotlarda (embriyonik yaşı 13,5 (E13,5); E18,5 ve postnatal 14 gün (P14)) incelenmesi amaçlandı. Çalışmada E13,5 embriyoların cinsiyeti SOX9 immünohistokimya işaretlemesiyle, E18,5 gonadların cinsiyeti H&E boyamasıyla, P14'lerin cinsiyeti ise makroskopik olarak tespit edildi. Cinsiyet tayininden sonra dişi ve erkek gonadların OTR ekspresyon seviyeleri immünohistokimya ve immünofloresan yöntemleri ile araştırıldı. Veriler istatistiksel olarak karşılaştırıldı. E13,5'a ait ovaryumdaki OTR+ hücreler organ boyunca daha diffüz bir şekilde görülürken, E13,5 testisteki OTR+ hücrelerin kümelenmiş bir şekilde olduğu anlaşıldı. E18,5 ve P14'e ait gonadlarda lokalizasyonuna göre OTR+ hücrelerin primordiyal folikül ve seminifer tübül yapılarının da ayırt edilmesiyle beraber bu alanlarda yoğunlaştığı görüldü. Bununla beraber hem ovaryum hem de testise ait bağ dokuda da OTR pozitifliği gözlemlendi. İkili immünofloresan işaretleme yöntemi ile işaretlenen immatür folikül ve immatür Sertoli hücrelerinin bir kısmının OTR eksprese ettiği görüldü. Dişide gelişimin ilerlemesiyle OTR eksprese eden tüm hücre sayısının oransal olarak azaldığı, OTR eksprese eden immatür folikül hücre sayısının ise arttığı tespit edildi. Erkekte ise bazı Sertoli hücreleri OTR eksprese ederken bazı Sertoli hücrelerinin etmediği görüldü. Sonuç olarak gelişimsel süreçte dişi ve erkek gonadlarda OTR ekspresyonu olduğu gösterilmiştir ve OTR ekspresyonunun gelişimsel olarak katkısı olabileceği düşünülmektedir.

Abstract:
The oxytocin receptor is the receptor for oxytocin which is a neuropeptide hormone that is expressed in various cells of ovarian and testicular tissue, as well as many tissues. There is limited information about the distribution of oxytocin receptors in male and female gonads during development. In this study, it was aimed to investigate the OTR expressions of immature gonad cells in the mouse reproductive system at different stages of development (embryonic age 13.5 (E13.5); E18.5 and postnatal day 14 (P14)). In the study, the sex of E13.5 embryos was determined by SOX9 immunohistochemical staining, the sex of E18.5 gonads was determined by H&E staining, and the sex of P14s was determined macroscopically. After sex determination, OTR expression levels of female and male gonads were investigated using immunohistochemistry and immunofluorescence methods. The data were statistically compared. OTR+ cells in the ovary of E13,5 were found to be more diffuse throughout the organ, while OTR+ cells in the E13,5 testis were found to be clustered. During the identification of primordial follicle and seminiferous tubule structures in E18,5 and P14 gonads, the OTR+ cells were observed to be more dense in  these areas. In addition, OTR positivity was observed The oxytocin receptor is the receptor for oxytocin which is a neuropeptide hormone that is expressed in various cells of ovarian and testicular tissue, as well as many tissues. There is limited information about the distribution of oxytocin receptors in male and female gonads during development. In this study, it was aimed to investigate the OTR expressions of immature gonad cells in the mouse reproductive system at different stages of development (embryonic age 13.5 (E13.5); E18.5 and postnatal day 14 (P14)). In the study, the sex of E13.5 embryos was determined by SOX9 immunohistochemical staining, the sex of E18.5 gonads was determined by H&E staining, and the sex of P14s was determined macroscopically. After sex determination, OTR expression levels of female and male gonads were investigated using immunohistochemistry and immunofluorescence methods. The data were statistically compared. OTR+ cells in the ovary of E13,5 were found to be more diffuse throughout the organ, while OTR+ cells in the E13,5 testis were found to be clustered. During the identification of primordial follicle and seminiferous tubule structures in E18,5 and P14 gonads, the OTR+ cells were observed to be more dense in  these areas. In addition, OTR positivity was observed in the connective tissue of both the ovary and testis. It was observed that some immature follicles and immature Sertoli cells stained by double immunofluorescence staining method, also expressed OTR. It was determined that as the development progressed in females, the number of all cells expressing OTR decreased proportionally, while the number of immature follicle cells expressing OTR increased. In male, it was observed that some Sertoli cells expressed OTR, while some Sertoli cells did not. As a result, it has been shown that OTR expression is present in both male and female gonads during development and it is thought that OTR expression may contribute to the developmental process.