KDM2B在小鼠卵母细胞及早期胚胎发育过程中的表达分析

Expression Analysis of KDM2B in Mouse Oocytes and Preimplantation Embryos

本研究旨在分析KDM2B在小鼠卵母细胞和早期胚胎中的表达规律,为KDM2B在卵母细胞减数分裂及胚胎发育过程中的生物学作用奠定基础。选择20只6～8周龄小鼠为试验动物,收集GV期、MⅡ期卵母细胞、2-细胞、4-细胞、8-细胞、囊胚各阶段胚胎,根据GenBank上已公布的小鼠(Mus musculus)KDM2B序列设计引物,采用实时荧光定量PCR(RT-qPCR)检测KDM2B在胚胎各阶段mRNA的表达水平;通过免疫荧光染色定位KDM2B蛋白在胚胎各阶段的分布。结果显示,GV期卵母细胞KDM2B mRNA的表达量极显著高于MⅡ期(P<0.01);在2-细胞、4-细胞和8-细胞mRNA表达量较低,囊胚期其表达量极显著升高(P<0.01);卵母细胞成熟过程中,KDM2B在GV期主要表达于细胞核中,MⅡ期卵母细胞核中的荧光信号极显著减弱(P<0.01),早期胚胎发育过程中,KDM2B蛋白在2-细胞、4-细胞、8-细胞胚胎中均不表达,囊胚期重新表达于细胞核。综上表明,本研究成功建立KDM2B在小鼠卵母细胞及胚胎细胞中的时空及时序表达模式,关于KDM2B参与调控减数分裂与胚胎发育过程具体的作用机制有待进一步研究。

The aim of this study was to investigate the expression pattern of KDM2B in mouse oocytes and preimplantation embryos,which provided important foundation for the mechanism study of KDM2B in meiosis and embryonic development.Twenty mice aged 6-8 weeks were selected as experimental material,GV and MⅡoocytes,2-cell,4-cell,8-cell,blastocyst were collected.The primer was designed based on Mus musculus KDM2B sequence in GenBank,and the expression pattern of KDM2B was detected by RT-qPCR in different stages of preimplantation embryos.Then immunofluorescence staining was used to locate the distribution of KDM2B protein in embryos of each period.The result showed that the mRNA expression level of KDM2B in GV oocyte was extremely significant higher than that in MⅡ(P<0.01).During the early embryonic development,the KDM2B was expressed in 2-cell to blastocyst,and extremely significant higher in blastocyst than that in other stages(P<0.01).In preimplantation embryos,KDM2B was observed in the nucleolar at GV oocyte stage,and fluorescence signatures in the nucleus clearly weakened at the MⅡstage(P<0.01),however KDM2B was not detected in 2-to 8-cell,and became detectable at blastocyst.This study established the spatial and temporal expression profiles of KDM2B,and the specific mechanism of KDM2B in regulating meiosis and embryonic development requires further investigation.