精原干细胞（SSCs）作为研究生殖细胞和精子发生的靶细胞，对转基因动物制备、解析生精障碍有重要意义。TET1作为表观修饰酶参与维持SSCs的自我更新，且TET1影响H3K27me3在靶基因启动子区的沉积。但TET1调控H3K27me3及H3K27me3维持SSCs自我更新的机制尚不清楚。本项目旨在TET1与组蛋白H3K27甲基化修饰角度分析SSCs自我更新。拟利用TET1过表达体外分离培养的SSCs，首先确认TET1通过JMJD3降低H3K27me3，维持SSCs自我更新能力；通过JMJD3及EZH2的修饰改变H3K27me3对靶基因的沉积，CHIP技术验证H3K27me3靶基因与下游通路；进一步通过磷酸化iTRAQ技术确定H3K27me3减少引起的差异蛋白与调控通路。阐明TET1-JMJD3-H3K27me3对SSCs自我更新的表观调控机制，为生精障碍的研究提供靶细胞及科学依据。

The spermatogonial stem cells (SSCs) are target cell for study of Germ cell and spermatogenesis, which is meaningful for analyzing the mechanism of spermatogenesis and treatment of male infertility. As an key epigenetic modified enzyme, TET1 (Ten-eleven translocation 1) plays important role in self-renewal of SSCs, which also reduces accumulation of H3K27me3. While the mechanism of TET1 and H3K27me3 in regulating SSCs’ self-renewal is still unclear. In our study, we assumes that TET1 reduces accumulation of H3K27me3 on promoters of target genes by upregulating the expression of JMJD3, and activate self-renewal related proteins and pathways to co-regulate SSCs’self-renewal. So, mouse SSCs are separated and cultivated in vitro are modified by lentiviral vector of TET1 to analyze the self-renewal ability and dynamic of H3K27me3. Then modification of JMJD3 and EZH2 to SSCs are used to analyze the dynamic of H3K27me3 and SSCs’self-renewal. Furthermore, iTRAQ phosphorylation technique is used to verify phosphoprotein and pathway effected by H3K27me3. These will help to verify epigenetic regulation mechanism of TET1-JMJD3-H3K27me3 to the self-renewal of SSCs, which will provide target cell and scientific evidence for revelation of dyszoospermia causes and regulation mechanism.