蛋白激酶C_ζ对小鼠受精卵发育早期基因组转录活化的影响

Effects of PKC_ζ on Early Embryonic Genome Transcription Activation in Mouse Eggs

为研究蛋白激酶Cζ (proteinkinaseCζ ,PKCζ)在小鼠受精卵细胞早期发育过程中对胚胎基因组活化影响 ,采用免疫印迹和细胞免疫荧光的方法 ,观察PKCζ的抑制剂对小鼠受精卵 1 细胞期G1和G2 不同时期小鼠受精卵基因组活化的影响 .小鼠 1 细胞期受精卵蛋白激酶C (PKC)的活性不断增加 ,并在G2 期达到最高 .PKC的抑制剂calphostinC可以明显抑制PKC的活性达 4 7% .同时calphostinC对受精卵 1 细胞期基因组的早期活化具有显著的抑制作用 (P <0 0 1) .在小鼠 1 细胞期受精卵的G2 期 ,具有活性的磷酸化PKCζ的含量明显多于G1期和卵母细胞MⅡ期 ,分别比它们高2 7%和 110 % .PKCζ的特异性抑制剂可以抑制受精卵 1 细胞期基因的转录和活化 (P <0 0 5 ) .实验结果表明 。

Effects of PKCζ on the activation of embryonic genome transcription in 1-cell stage fertilized mouse eggs were explored. The effects of PKC antagonist calphostin C and PKCζ specific inhibitor on the activation of embryonic early transcription were observed by Western blotting and cell immunofluorescence. PKC activity elevated gradually from G 1 phase to late G 2 phase in fertilized 1-cell stage eggs, and increased to a maximum in G 2 stage. Calphostin C, a PKC inhibitor, inhibited the PKC activity about 47% in 1-cell stage eggs. Calphostin C inhibited the early transcription in 1-cell stage eggs(P<0.01). PKCζ-Thr410 increased in G 2 phase accompanying transcription activation. The relative quantities of PKCζ-Thr410 in G 2 phase were about 27% and 110% higher than those in G 1 phase in 1-cell stage eggs and MII oocyte, respectively. PKCζ specific inhibitor can also inhibit the early transcription in 1-cell stage eggs(P<0.05). The results suggest PKCζ participates into early transcription activation in mouse 1-cellstage eggs.