Tdrd9调控雌雄同体石斑鱼原始生殖细胞发育的分子机制研究

雌雄同体鱼类性别分化的分子机制一直是科学界的未解之谜，而原始生殖细胞的性别选择机制是该问题的核心。此前我们发现tdrd9基因在原始生殖细胞(PGC)中存在特异性表达。解析tdrd9在PGC发育过程中的分子调控机制，有助于深入阐述石斑鱼PGC发育的分子机制。本项目以我国重要海水经济鱼类斜带石斑鱼为研究对象，采用荧光定量PCR、原位杂交和免疫组化等方法，分析tdrd9在斜带石斑鱼PGC发育阶段的表达和定位情况；利用RNAi技术，在个体中解析tdrd9对PGC发育的调控作用；采用免疫共沉淀技术，结合piRNA测序分析Tdrd9与Piwi蛋白的相互作用以及共同调控的piRNA。本项目旨在明确斜带石斑鱼tdrd9的动态表达规律，阐明tdrd9调控PGC发育的分子作用机制，为探究石斑鱼性别分化分子机制提供新的思路，从而为石斑鱼类育种和遗传改良提供理论基础。

The molecular mechanism of sex differentiation in hermaphrodite fishes has been an unsolved mystery in science, and the mechanism of sex selection in primordial germ cells (PGCs) is the core of this problem. Previously, we found that tdrd9 was expressed in PGC specifically. Analyzing the molecular regulation mechanism of tdrd9 in the development of PGC will help us to explain the molecular mechanism of PGC development of grouper further. In this project, our subject is orange-spotted grouper, an important marine economic fish in China. Fluorescence quantitative PCR, in situ hybridization, immunohistochemistry and other technologies are used to explore the expression pattern of tdrd9 in development of PGC in orange-spotted grouper. RNAi techniques are conducted to analysis the regulation of tdrd9 on the development of PGC in individual. The interaction between Tdrd9, Piwi proteins and piRNAs is analyzed by immunoprecipitation technique combined with piRNA sequencing. The project aims to clarify the dynamic expression rule of Tdrd9 in orange-spotted grouper, clarify the molecular mechanism of Tdrd9 regulating the development of PGC, and provide a new idea for exploring the molecular mechanism of gender differentiation in grouper, so as to provide a theoretical basis for grouper breeding and genetic improvement.