小鼠受精卵早期发育过程中PKC对cdc2和cdc25C活性的影响

为研究小鼠受精卵细胞早期发育过程中PKC对cdc2和cdc2 5C活性的影响 ,采用免疫印迹和电泳迁移率差异分析的方法 ,观察PKC的激活剂TPA及其抑制剂星形孢子素对小鼠受精卵一细胞期cdc2和cdc2 5C活性的影响 .10nmol L的TPA作用 10min后 ,小鼠受精卵一细胞期卵裂率明显大于对照组 (P <0 0 5 ) ,而星形孢子素作用后卵裂率显著下降 (P <0 0 1) .TPA处理后 ,受精卵中呈去磷酸化状态的活性cdc2明显增加 ,没有活性呈磷酸化状态的cdc2 5C明显减少 ;而星形孢子素处理的受精卵中没有活性的cdc2明显增加 ,有活性的cdc2 5C明显减少 .结果表明 ,TPA短时间作用可以促进小鼠一细胞期受精卵分裂 ,星形孢子素抑制受精卵的分裂 ;TPA可以促进cdc2的去磷酸化以及cdc2 5C的磷酸化 ,从而促进G2 M转换 ,星形孢子素则抑制cdc2和cdc2 5C的活性 ,阻止受精卵由G2 期进入M期

The Effects of PKC on the Activation of cdc2 and cdc25C in One Cell Stage Fertilized Mouse Eggs

The effects of PKC on the activation of cdc2 and cdc25C in one cell stage fertilized mouse eggs were explored. The effects of PKC agonist TPA or its antagonist staurosporine on the activation of cdc2 and cdc25C were observed by immunoblotting and electrophoresis mobility shift assay. The division percentage of one cell stage eggs increased significantly vs. control after the treatment of 10 nmol/L TPA 10 min ( P <0 05) and decreased after the treatment of staurosporine ( P <0 01). After treatment with TPA, the amount of phosphorylated activated cdc2 increased obviously and that of dephosphorylated inactivated cdc25C was decreased in one cell stage eggs. The amount of inactivated cdc2 increased and that of activated cdc25C decreased in one cell stage eggs if it was treated with staurosporine. The results suggest that TPA treatment in a short time could accelerate cleavage of one cell stage eggs, while staurosporine inhibit cleavage. TPA could activate cdc2 and cdc25C and promote G2/M transition and staurosporine had the opposite effect.