USP9Y在雄性不育小鼠中的作用和分子机制的研究

男性生殖功能障碍导致不育的研究是目前生殖领域的研究热点之一。研究男性不育的致病机理，阐明精子发生异常的调控机制，将为男性不育的治疗及探索男性不育治疗新途径奠定理论基础。我们利用基因芯片技术的前期研究结果表明，在Agtpbp1蛋白表达缺失的雄性不育的pcd小鼠精巢中USP9Y mRNA表达水平反而增加，因此本课题利用免疫组化、western blot和荧光定量PCR等技术分析USP9Y在小鼠精巢中的表达，结合免疫共沉淀技术研究Agtpbp1和USP9Y之间的相互作用和调控关系，探讨USP9Y过度表达对细胞周期和凋亡的影响。同时采用GST affinity purification的方法并结合质谱技术，获取USP9Y蛋白相互作用组（interactome）图谱，结合生物信息学资源，阐明USP9Y和其相互作用蛋白在精子发生异常中的分子机制，以期为治疗人类男性不育疾病提供扎实的理论基础

Male reproductive dysfunction that leads to sterility is currently a hot topic in the field of reproduction. Elucidation of the pathogenic and regulatory mechanisms of male infertility and sperm abnormalities will provide the theoretical foundation for male infertility treatment and exploration of new approaches to the treatment. Our previous research results by DNA microarray analysis showed that USP9Y mRNA expression level was increased in the testes of sterile pcd males, in which Agtpbp1 was mutated. Therefore this project aims to analyze the expression of USP9Y in the testes of pcd males at different development stages by immunohistochemistry, Western blot and real-time PCR methods. The interaction and regulation between Agtpbp1 and USP9Y proteins will be studied using immunoprecipitation and Western blot, respectively. The effects on the cell cycle and apoptosis by USP9Y overexpression will be explored further. At the same time we will use GST affinity purification method combined with mass spectrometry to obtain the USP9Y protein interactome. Together with the results by the informatics analysis and further experimental study, elucidation of the USP9Y and its interacting proteins in sperm abnormalities and the molecular mechanisms underlying it will positively provide a solid theoretical foundation for the treatment of human male infertility diseases.