Prmt5在鱼类胚胎和生殖细胞发育中的作用机理研究

项目名称： Prmt5在鱼类胚胎和生殖细胞发育中的作用机理研究

项目编号： No.31272645

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 赵浩斌

作者单位： 华中师范大学

项目金额： 80万元

中文摘要： Prmt5是蛋白质精氨酸甲基化酶，在多种生物功能如RNA剪切、肌肉发生、细胞凋亡、哺乳动物干细胞维持和生殖细胞发生等过程中发挥重要作用。鱼类Prmt5在其胚胎及生殖细胞发育中的功能研究鲜有报道。本研究以青鳉为模式对象,应用蛋白组学方法,研究胚胎发育关键阶段和性腺的蛋白质甲基化状况。通过基因敲降和过量表达，观察Prmt5对胚胎发育、生殖细胞发生的作用以及Prmt5对细胞增殖和凋亡的影响，并通过转录组分析了解Prmt5对胚胎基因表达的影响。采用酵母双杂交和BiFC、免疫共沉淀等方法研究与Prmt5相互作用的其它蛋白质。对一些生殖细胞和干细胞特异基因通过原位杂交、定量RT-PCR等方法做进一步研究。项目的完成将对Prmt5在鱼类胚胎发育和生殖细胞发生的作用有比较全面而深入的了解，更深刻地了解甲基化在鱼类发育中的调控作用。本项目的研究成果对水产养殖鱼类生殖繁育甚至干细胞培育都具有参考作用。

中文关键词： 青鳉；Prmt5；互作蛋白；胚胎发育；基因表达

英文摘要： Prmt5, a protein arginine methyltransferase, plays important roles in multi bioproccesses such as RNA splicing, myogenesis, cell apoptosis, germ cell development and stem cell maintenance in mammals. However, few studies concern about the functions of Prmt5 in embryonic and germ cell development of fish. We will study the post-translational modification of proteins with symmetrical dimethylated arginines (sDMA) in the embryos and gonads of medaka by using proteomic analysis, the roles of Prmt5 in embryonic and germ cell development including cell proliferation and apoptosis by means of gene knockdown and overexpression, the effects of Prmt5 on gene expression in embryos by transcriptomic analysis, and the proteins that interact with Prmt5 by using yeast two hybridization, bimolecular fluorescence complementation and co-immunoprecipitation. The genes, which are specific to germ cells or stem cells, will be studied further by using in situ hybridization and quantitative RT-PCR, etc. We will get a full and extensive understanding of the roles and mechanisms of Prmt5 on embryonic and germ cell development in fish and the regulation of post-translational methylation on fish development after finishing this project. The results obtained from this study will be used as references for reproduction and cultivation and s

英文关键词： Medaka；Prmt5；interacting protein；embryonic development；gene expression