中文摘要
细胞凋亡是影响生殖的重要因素,卵泡刺激素(FSH)对卵泡细胞凋亡或闭锁有明显调节作用,但 FSH对卵泡细胞凋亡和凋亡相关基因及FSHR表达的作用和调控机理在兰州大尾羊至今无研究报道。本项目以兰州大尾羊为研究对象,利用免疫组织化学、TUNEL分析、原位杂交、RT-PCR、Western blot、流式细胞仪、Annexin-V分析、显微和超微组织学等方法,通过在体和离体研究,(1)探讨不同发育阶段卵泡中FSH与FSHR结合对细胞凋亡的促进或抑制作用;(2)研究卵泡细胞凋亡和凋亡相关基因(Bcl-2/Bax, Fas/FasL, PTK-STAT, Caspase-3,Caspase-9等)表达及相互关系, 分析不同浓度FSH对卵巢FSHR基因和FSHR蛋白表达的调控作用;(3)阐明FSH对兰州大尾羊卵泡发育过程中细胞凋亡的调控机制和通路,为提高兰州大尾羊卵巢储备能力和繁殖性能提供技术依据。
英文摘要
A large number of follicles happen the apoptosis and atresia during the follicular development. The follicular atresia are caused mainly by cell apoptosis. The apoptosis is an important factor affecting the female reproductive functions. Follicle-stimulating hormone (FSH) inhibits or promotes the follicular apoptosis or atresia by binding to its receptor subtypes (FSHRα/FSHRβ). However, this actions of FSH has not been reported in Lanzhou fat-tailed sheep, especially in aspects of the ovaries FSH receptors (FSHR) expression and the mechanism of follicular cell apoptosis.The aims of the present project are as followings. (1) to explore the inhibition or promotion effects of FSH on the follicle apoptosis in the different follicle stages by combining FSHR; (2) to investigate the regulatory effects of FSH on the FSHR gene and FSHR protein expressions in the ovaries of Lanzhou fat-tailed sheep in vivo and in vitro conditions, as well as the correlations between the follicle apoptosis and apoptosis-related gene (Bcl-2/Bax, Fas/FasL, PTK-STAT, Caspase-3, Caspase-9) using immunohistochemistry, TUNEL assay,hybridization in situ , RT-PCR, Western blot, flow cytometry, Annexin-V assay and microscopy histology methods; (3) to demonstrate the FSH modulation mechanisms on follicle apoptosis during the follicular development in Lanzhou fat-tailed sheep. So as to enhance ovarian reserve capacity and reproductive performance for Lanzhou fat-tailed sheep, also to provide new ideas.
