关键词: Wee1B, 短发夹RNA, 有丝分裂, 受精卵, 小鼠

Abstract:

To study the influence of protein kinase Wee1B in the early development of mouse embryos by the means of specific Wee1B shRNA.Methods The mouse one-cell-stage embryos were collected after superovulation,and microinjected with TE buffer,control shRNA or Wee1B shRNA respectively,then the specificity of Wee1B shRNA was examined by the methods of RT-PCR and Western blotting.After RNA interference,the morphological changes and cleavage rate of mouse embryos were observed under fluorescence microscope,and the Wee1B activity of mouse embryos was detected by autoradiography.
Results Compared with TE buffer and Control  shRNA groups,the expression levels of Wee1B mRNA and protein  in Wee1B shRNA group were significantly reduced,and the activity of Wee1B was also decreased.The cleavage rates of mouse one-cell-stage embryos in non-injection,TE buffer,Control shRNA groups 33 h after hCG injection were 71.91 %±1.74%,72.90 %±2.25%, and 72.28 % ±2.06%,and there was no obvious difference among the three groups(P>0.05); the cleavage rate in Wee1B shRNA group  was  90.00%±0.77% at 33 h,which was markedly higher than those of the controls (P<0.01).Conclusion Wee1B shRNA can promote the  onset of mitosis of mouse one-cell-stage embryos,suggestting that Wee1B negatively regulates mitosis during early embryogenesis in mouse one-cell-stage embryos.

Key words: Wee1B；shRNA；mitosis；embryos, mice