斑马鱼精巢特异表达基因lrp2bp对雄激素11-KT合成及分泌的调控机制研究

Until now the underlying mechanism of 11-KT synthesis and secretion in fish remains largely unknown. In previous studies, we cloned a gene specifically expressed in the zebrafish testis, lrp2bp. In situ hybridization and immunohistochemistry experiments showed that lrp2bp is specifically expressed in Leydig cells. In lrp2bp mutants, the serum 11-KT content was significantly decreased. This project intends to fully investigate the expression characteristics and subcellular localization of lrp2bp protein during zebrafish development to speculate the possible function and mechanism of lrp2bp during development. By analyzing the phenotypes of lrp2bp-/- mutants, transcriptome sequencing and quantitative PCR validation, the lrp2bp downstream genes were screened out, combined with a dual luciferase reporter system and co-immunoprecipitation technology to resolve zebrafish lrp2bp in 11-KT. The position in the synthetic and secretory signaling pathways ultimately reveals the biological function of lrp2bp in gonad development in zebrafish. This study will further deepen our understanding of the molecular mechanism of 11-KT synthesis and secretion in fish. To reveal the gene regulatory networks related to the synthesis and secretion of zebrafish 11-KT and its mechanism of action will laid the theoretical foundation for the artificial breeding and variety improvement of cultured fish.

目前关于鱼类雄激素合成及分泌的调控机制仍有待完善。我们前期在斑马鱼中克隆到精巢组织特异表达基因lrp2bp，经原位杂交和免疫组化实验显示lrp2bp特异性地表达于精巢间质细胞。敲除lrp2bp后，血清中雄激素11-KT的含量显著降低。本项目拟通过完善lrp2bp基因在斑马鱼个体发育过程中的表达特征和亚细胞定位，以推测lrp2bp在发育过程中可能的功能及作用机制；通过分析lrp2bp-/-突变体的表型、转录组测序和荧光定量PCR验证，筛选出lrp2bp下游基因，同时结合双荧光素酶报告系统和免疫共沉淀技术解析斑马鱼lrp2bp在11-KT合成及分泌信号通路中的地位，最终揭示lrp2bp在斑马鱼性腺发育中的生物学功能。本研究将进一步加深人们对鱼类11-KT合成及分泌分子机制的认识，揭示斑马鱼11-KT合成及分泌相关的基因调控网络及其作用机理，为养殖鱼类的人工繁育和品种改良奠定理论基础。