核膜孔亚复合物Nup98/Rae1对小鼠卵母细胞体外减数分裂成熟的影响OA北大核心CSTPCD

[Objective]The objective of this experiment was to explore the effect of nucleoporin 98(Nup98)and ribonucleic acid export 1(Rae1)on meiotic maturation of mouse oocytes in vitro.[Method]The oocytes of 4-week-old SPF female Kunming mice were isolated and cultured in vitro.The expressions of Nup98 and Rae1 genes in mouse oocytes were detected by Real-time quantitative PCR.The colocalization of Nup98 and Rae1 was detected by immunofluorescence.The Nup98 and Rae1 genes in mouse oocytes were knocked down by electrotransfer siRNA interference technology.The interference efficiency was detected by Real-time quantitative PCR,and germinal vesicle breakdown(GVBD)and first polar body extruction(PBE)were observed.The spindle assembly and chromosome arrangement of mouse oocytes after Nup98 and Rae1 genes knocked down were detected by immunofluorescence method.The rate of chromosome aneuploidy was detected by chromosome climbing film technique.[Result]Both Nup98 and Rae1 genes were expressed in mouse oocytes.During the germinal vesicle(GV)stage,Nup98 and Rae1 were co-located at the nuclear membrane margin.During meiosis,Nup98 and Rae1 were concentrated on the kinetosomes of chromosomes.The results of siRNA interference test showed that,compared with control group,knockdown of Nup98 and Rae1 gene alone had no significant effect on the expression of the other gene(P＜0.05).After double-knockdown of Nup98 and Rae1 genes,there was no significant effect on the meiosis recovery rate of oocytes(P＞0.05),the PBE rate of mouse oocytes was extremely significantly increased after 9.5 h development(P＜0.01),and the chromosome separation rate and aneuploidy rate were extremely significantly or significantly increased(P＜0.01 or P＜0.05).[Conclusion]The Nup98 and Rae1 genes were involved in chromosome separation during mouse oocyte meiosis,influencing the occurrence of aneuploidy and thus the developmental potential of fertilized embryos.