项目名称： 中华绒螯蟹GnRH信号系统的鉴定及其在卵母细胞成熟中的调控作用

项目编号： No.41476130

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 天文学、地球科学

项目作者： 邱高峰

作者单位： 上海海洋大学

项目金额： 82万元

中文摘要： 促性腺激素释放激素（GnRH）在脊椎动物生殖调控中起核心作用，但在虾蟹类等无脊椎动物中相应的激素信号系统尚未建立，本课题组之前分离鉴定了中华绒螯蟹GnRH，同时证明该激素能够刺激卵母细胞生发泡破裂、促进卵母细胞减数分裂成熟，本研究拟进一步利用转录组学、基因芯片分析方法筛选鉴定脑、胸神经团、卵巢组织GnRH效应基因，推导潜在的GnRH信号通路；利用荧光定量PCR、原位杂交、免疫荧光定位等方法分析GnRH受体等关键通路基因的时空表达谱，探究基因的表达与卵母细胞成熟的关系；利用蛋白相互作用、RNAi实验进一步验证GnRH信号通路，解析GnRH信号系统在调控卵母细胞成熟中的作用，蟹类GnRH信号系统的揭示对于解明卵母细胞成熟机制和无脊椎动物GnRH信号系统的进化具有重要学术价值。

中文关键词： 中华绒螯蟹；促性腺激素释放激素；信号系统；卵母细胞成熟

英文摘要： Vertebrate gonadotropin-releasing hormone (GnRH) plays a central role in the regulation of reproduction. However, so far the correspondent GnRH signaling system has not been characterized in invertebrates including shrimps and crabs. In our previous studies, we isolated a GnRH homolog from Chinese mitten crab Eriocheir sinensis. In vivo injection experiments demonstrated that the crab GnRH can induce germinal vesicle breakdown (GVBD) and meiotic maturation of oocytes. To further explore the potential GnRH signal pathway and insight into its role on regulation of the oocyte maturation, firstly we will perform comparative analysis on the transcriptom of the brain, thoracic ganglion,and ovary to isolate the genes in response to the crab GnRH treatment and deduce the GnRH signal pathway. By means of real-time PCR, in situ hybridization and immunofluoresence, then we will examine the spatio-temporal expression profiles of GnRH-responsed genes including GnRH receptor on both mRNA and protein levels to provide information on the possible function of those genes during oocyte maturation. Finally, the exact GnRH signal pathway will be characterized by proteins interaction and RNAi approach. The present studies will further our understanding of regulatory mechanism of oocyte maturation in the decapod species and the evolution of GnRH signal system in invertebrates.

英文关键词： Chineses mitten crab;GnRH;signaling system;oocyte maturation