【作者】 付伟； 赵咏梅； 武迪迪； 冯晨； 宗志宏； 王亚杰； 刘莹； 刘奕； 于爱鸣； 陈菲； 于秉治；

【Author】 FU Wei, ZHAO Yong-mei, WU Di-di, FENG Chen, ZONG Zhi-hong, WANG Ya-jie, LIU Ying, LIU Yi, YU Ai-ming, CHEN Fei, YU Bing-zhiDepartment of Biochemistry, School of Basic Medical Science, China Medical University, Shenyang 110001, China, E-mail: ybzbiochem@yeah.net

【机构】 中国医科大学基础医学院生物化学教研室；

【摘要】 为研究蛋白激酶Cζ(protein kinase Cζ,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达到最高。PKC的抑制剂calphostin C可以明显抑制PKC的活性达47%。同时calphostin C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<0.01)。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27%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<0.05)。实验结果表明,PKCζ参与了小鼠受精卵基因组早期转录的调控。更多还原

【Abstract】 Effects of PKCζon the activation of embryonic genome transcription in 1-cell stage fertilized mouse eggs were explored. The effects of PKC antagonist calphstin C and PKCζ specific inhibitor on the activation of embryonic early transcription were observed by Western blotting and cell immunofluorescence. PKC activtity elevated gradually from Gl phase to late G2 phase in fertilized 1-cell stage eggs, and increased to amaximum in G2 stage. Calphostin C, a PKC inhibitor, inhibited the PKC activtity 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 G2 phase accompanying transcription activation. The relative quantities of PKCζ-Thr410 in G2 phase were about 27% and 110% higher than those in Gl phase in 1-cell stage eggs and M II oocyte, respectively. PKCζspecific 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-cell stage eggs.更多还原