染色体分离异常引起的非整倍体是导致胚胎发育异常、流产等生殖疾病的主要原因。申请人利用新发现的突变体RanT24NT42A抑制Ran GTP后，引起卵母细胞提前进入减数分裂后期，染色体分离异常。据此推测，Ran GTP可能通过调控APC/C（后期促进复合物/细胞周期体）影响减数分裂周期进程，但调控机制尚不清楚。本项目拟：①利用活细胞成像技术，明确Ran GTP对卵母细胞成熟的影响；②检测APC/C及周期相关蛋白表达变化，分析Ran GTP调控APC/C功能和染色体分离的机制；③抑制Ran GTP介导的出入核过程，并通过蛋白质谱挖掘Ran GTP调控APC/C功能的关键蛋白；④利用免疫共沉淀和Trim-Away技术，验证关键蛋白调控APC/C的功能，构建APC/C调控卵母细胞成熟的信号通路。本项目预期阐明Ran GTP调控APC/C功能和染色体分离的分子机制，为改善卵子质量提供理论依据。

Aneuploidy caused by chromosomes mis-segregation is the main reason of abnormal embryonic development and abortion. Our previous work found that inhibition of Ran GTP by RanT24NT42A accelerated the meiotic anaphase occurrence and caused the abnormal chromosomes segregation during oocyte maturation. Therefore, we speculate that Ran GTP could regulate the cell cycle progress through mediating the APC/C (anaphase promoting complex/cyclosome) function, but the regulatory mechanism is still unclear. In this project we carry out the following studies: firstly, live cell imaging is used to determine the effect of Ran GTP on oocyte maturation. Secondly, we investigate the protein level of APC/C and cell cycle-related proteins, and explore the regulating mechanism of Ran GTP in APC/C function and chromosomes segregation. Thirdly, the proteomics and mass spectrometry technology is used to determine the potential target that regulates APC/C function after inhibition of Ran GTP-mediated nucleocytoplasmic transport. Fourthly，the CO-IP and Trim-Away technologies are used to verify the effect of potential target on APC/C function, which will build the signal pathway of Ran GTP-regulated APC/C function. This project expects to clarify the regulating mechanism of Ran GTP in APC/C function and chromosomes segregation, that will provide a theoretical basis for improving oocyte quality.