虾蟹类性别决定和性分化机制是甲壳动物发育生物学研究的一个热点。三疣梭子蟹单雌化养殖能显著提升其人工养殖的经济效益。胰岛素样促雄性腺激素（IAG）基因被认为在虾蟹类雄性分化和雄性性征的发育上起关键的调控作用。在已经掌握三疣梭子蟹促雄性腺组织学特征、克隆IAG全长cDNA序列并分析其组织差异表达的基础上，拟进一步综合应用RNAi技术、活体注射重组蛋白（rIAG）和离体培养等方法阐明IAG基因对性腺发育、配子发生、第二性征发育、蜕皮和生长等生理过程的调控作用，确定IAG基因调控三疣梭子蟹雄性分化的时间窗口；克隆分析IAG基因启动子和表达调控序列，揭示转录因子、miRNAs、神经递质和CHH家族神经肽对IAG基因表达的调控机理。开展本项研究，将为实现三疣梭子蟹性反转和单雌化养殖提供理论指导，还将有助于阐明其性别决定和分化机制。

The mechanism of sex determination and differentiation in decapods is a hot research topic for crustacean developmental biology. The hologynic culture of the swimming crab Portunus trituberculatus is provided with great economic values. It is well known that the insulin-like androgenic gland hormone (IAG) gene plays a crucial roles on male sex differentiation and sexual characteristics development of decapods. Based on a well understanding of the histology of androgenic gland of P. trituberculatus, and the preliminary result of the cDNA cloning and tissue distribution of IAG mRNA, this project aims to illustrate the regulatory roles of IAG gene on gonadal development, gametogenesis, secondary sexual characteristics development, molting and growth, and to confirm the time window of IAG gene in controlling the male sexual differentiation of P. trituberculatus, with the integrated utilization of RNA interference, in vivo injection of recombinant protein of IAG, and in vitro incubation. In addition, the project will aim to identify the 5’-flanking sequence of IAG gene, analyze its transscription factor binding sites and the promoter transcriptional activity, and to reveal the mechanism of transcription factors, miRNAs, neurotransmitters, and CHH family neuropeptides in regulation of IAG gene expression. To carry out this research will provide theoretical guidance for sex reversion and all-female population culture of P. trituberculatus, and help to elucidate the sex determination and differentiation mechanism of this species.