ETV5介导的小鼠精原干细胞自我更新转录调控机制研究

项目名称： ETV5介导的小鼠精原干细胞自我更新转录调控机制研究

项目编号： No.31271581

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 生物科学

项目作者： 吴鑫

作者单位： 南京医科大学

项目金额： 75万元

中文摘要： 精原干细胞(SSCs)是雄性生精基础。SSCs体内自我更新、体外培养和增殖依赖于GDNF。我们前期的研究提示转录因子蛋白ETV5，不仅自身表达受GDNF影响，同时还在转录水平上调控GDNF下游基因，如Bcl6b、Lhx1、Brachyury(T)、Cxcr4等，在SSCs自我更新的过程中起关键作用。此外，我们发现，从小鼠到人类，物种间有七千多万年进化差距，但其SSCs的自我更新机制具有保守性，ETV5在这两个雄性物种早期生殖细胞中都高表达，提示它在SSCs中的作用可能也具有保守性。本项目通过染色体免疫沉淀方法，结合新一代DNA深度测序技术，探讨SSCs中受到ETV5调控的下游基因；并通过RNA干扰，慢病毒转染等方法，研究这些下游基因表达的改变是否会影响到SSCs增殖、迁移、分化以及再生精等生物学功能，从而在分子水平阐明雄性生殖干细胞的调控机制，为个体遗传信息稳定传递提供理论依据。

中文关键词： 精原干细胞；雄性不育；ETV5；长链非编码RNA；磷酸化组学

英文摘要： Spermatogonial stem cells (SSCs) serve as the foundation of spermatogenesis. In vitro growth of SSCs is dependent on the growth factor GDNF. We have identified a critical transcriptional factor, ETV5 not only as a downstream regulator of GDNF signaling that mediated the expression of several known SSC self-renewal related genes, including Bcl6b and LIM homeobox 1 (Lhx1), but also as a regulator of Brachyury (T) and CXC chemokine receptor type 4 (Cxcr4), which is of biological importance and functions to promote self-renewal or differentiation of mouse SSCs cultured in vitro. Our previous data indicated that remarkable conservation of gene expression, especially for self-renewal genes, in these prepubertal germline cells between mouse and human that diverged phylogenetically ~75 million years ago. Notably, Etv5 was also found greatly elevated in the early stage of male germline cells in both species, which indicates the regulation of Etv5 could be essential in SSCs. In current study, we would combine chromatin immunoprecipitation (CHIP) and next-generation high throughput sequencing to explore ETV5 mediated regulation in SSCs, together with stem cell culture, small RNA interference, lentivirus transfection and in vivo transplantion. We will also analyze the SSC growth dynamics, SSC migration and SSC differentiati

英文关键词： Spermatogonial stem cell；male infertility；ETV5；lncRNA；phosphoproteomic