【摘要】 目的了解印迹基因Snrpn在人类卵母细胞和植入前胚胎的表达,以分析Snrpn基因表达对人类卵母细胞和早期胚胎发育的影响。方法采用单细胞巢式逆转录多聚酶链式反应(Single Cell Nested RT-PCR)方法,检测人类卵子和植入前各个阶段胚胎的印迹基因Snrpn mRNA表达情况。结果Snrpn在人类卵母细胞GV、MI、MII和植入前胚胎2、4、6、8细胞期都有表达,单卵母细胞扩增成功率为87.5%,单个胚胎为75.8%,平均扩增成功率为79.1%。单个卵母细胞和单个卵裂球扩增成功率无显著性差异(P>0.05),但单卵母细胞较单个卵裂球高。结论建立了比较稳定、可靠的单细胞巢式RT-PCR技术检测基因表达。印迹基因Snrpn在人类卵细胞和植入前胚胎的表达对卵子生长和胚胎发育有着重要意义,为早期胚胎阶段该基因表达正常图谱提供实验依据更多还原

【Abstract】 Objective To investigate expression of imprinted gene SNRPN in human oocytes and preimplantation embryos and the effect of SNRPN gene expression on the development of human oocytes and embryos.Methods We tested the expression of imprinted gene SNRPN in human oocytes and preimplantation embryos by use of single cell nested reverse transcription-polymerase chain reaction(RT-PCR) amplification.Results SNRPN was expressed in human oocytes of different developmental stages [germinal vesicle(GV),metaphase I(MI) and metaphase II(MII) oocytes] and in preimplantation embryos(2cell,4-cell,6-cell,8-cell phase).The successful rate of amplification in oocytes and embryos was 87.5% and 75.8%,respectively,with average successful rate being 79.1%.There were no significanct differences in successful rate of amplification between single oocyte and individual embryo(P>0.05),but the successful rate of amplification was higher in the former than in the latter.Conclusion We developed a comparatively stable and reliable technique of single cell nested RT-PCR to test the expression of genes.Expression of imprinted gene SNRPN plays an important role in human oocyte growth and preimplantation embryo development.It also provides an experimental basis for normal map of expression of SNRPN gene at the early embryo stage.更多还原