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

项目名称： 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

项目编号： No.31472087

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 畜牧学与草地科学

项目作者： 李碧春

作者单位： 扬州大学

项目金额： 85万元

中文摘要： 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

中文关键词： 鸡；基因功能；生殖干细胞；信号通路；甲基化修饰

英文摘要： 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.

英文关键词： chicken;gene function;germ-line stem cells;signaling pathway;Methylated modification