ESCs向雄性生殖干细胞分化的特异表达新候选基因作用机制研究

Spermatogonial stem cells(SSCs)could be used to replace embryonic stem cells(ESCs) for disease therapy and genetic modification since it has the merits of pluripotency and do not involve in anyethical and immune rejection problems, therefore, it caused great concern in the biological community. However, the mechanism of germ cells differentiation is still unclear, and the interaction between the key genes and inducer is also unexplained. All these problems result in the inefficient induction, and thus, it is very hard to obtain fully functional germ cells in vitro and could not meet the requirements of research and practice. And the related research of poultry are still less less. Therefore, to establish the induction system in vitro, it is very necessary to deeply know about the process and mechanism of the male germ cells fate determination.Based on the previous study about the screening the key genes and signaling pathways involved in the germ cells differentiation,this project is aimed to confirm the regulation mechanism and expession pattern of the new candidate genes gotten by DGEs. A clear relationship between these candidate genes and signaling pathway will be described.The internaction and the regulation pattern between the node genes and new cnadidate genes will be detected by protein interaction technology. The candidate genes methylation and its effection on the fate of SSCs will be proven. This study will provide a theoretical basis for clarifying the differentiation mechanism of germ cells, and supply a strong practical technical guidance for improving the induction efficiency of germ cells.

精原干细胞(SSCs)的可塑性使其能替代胚胎干细胞用于治疗和遗传修饰，而不涉及伦理和免疫排斥问题，为其应用开辟了广阔前景。但是调控生殖干细胞分化发生的关键基因及其作用机理仍不清楚，导致干细胞向SSCs诱导分化效率低和重复性差，难以获得能满足研究和实践需要的大量生殖干细胞，而家禽的有关研究报道更少。所以在成熟的生殖细胞体外诱导分化和体内分离系统建立之前，深刻认识生殖干细胞分化发生的进程及机制显得非常迫切和必要。本项目旨在前期以家鸡为模式动物筛选了参与雄性生殖干细胞发生的关键基因和信号通路基础上，探悉新发现的特异表达候选基因作用机制。弄清它们的表达规律和作用；明确候选基因与信号通路的关系；利用蛋白质相互作用技术研究信号通路的节点基因与候选基因互作对SSC命运决定的调控机制；探明候选基因甲基化修饰对SSC命运决定的影响。为阐明生殖细胞的发生机制提供理论基础，为提高生殖细胞诱导效率提供实践技术支撑

按照项目申请书及合同书的计划，2015年度-2018年度该课题的研究计划要点是：（1）鸡原始生殖细胞形成过程中C2EIP功能及其调控机制的研究；（2）鸡原始生殖细胞（PGCs）生成过程中C1EIP基因调控机制的研究；（3）组蛋白乙酰化协同Sox2转录因子调控Cped1促进鸡SSCs形成的研究；（4）HoxA5转录因子调控LBC基因促进鸡ESCs向SSCs分化的研究。该课题的研究按照合同书计划内容正常有序地进行，并完成全部研究计划。