小鼠受精卵早期发育过程中PKA在M/G1期进程中的作用  

EFFECT OF PROTEIN KINASE A ON M/G1 TRANSITION IN EARLY DEVELOPMENT OF MOUSE FERTILIZED EGGS

作　　者：王亚杰[1] 刘莹[1] 陈菲[1] 于爱鸣[2] 宗志红[2] 于秉治[1] 

机构地区：[1]中国医科大学基础医学院生物化学教研室 [2]中国医科大学基础医学院病理生理教研室,沈阳110001

出　　处：《细胞生物学杂志》2003年第3期174-178,共5页Chinese Journal of Cell Biology

基　　金：国家自然科学基金重点课题(39730460);国家重点基础研究发展规划(973)项目(G1999055900-2)资助

摘　　要：为研究小鼠体内1-细胞期受精卵蛋白激酶A(PKA)对M/G1期进程的影响,应用热稳定性抑制剂PKI显微注射入1-细胞期受精卵内,观察M期促进因子(MPF)及PKA活性变化以及MPF调节亚基Cyclin B含量情况。发现PKI显微注射后PKA活性低,而MPF活性在hCG后27.5 h即达高峰,较对照组提前30分钟。PKI达一定浓度则MPF活性不下降,出现M/G1阻滞;与此同时Western blotting法显示PKI注射后Cyclin B含量在M末期相当于M中期水平。结果表明,PKI显微注射抑制PKA活性后MPF活性呈高峰值,高浓度PKI显微注射可引起M/G1阻滞,其机制与PKI干扰了Cyclin B降解有关。In order to investigate the effect of Protein Kinase A (PKA) on transition from mitosis to interphase in mouse fertilized eggs, PKA inhibitor PKI (protein kinase inhibitor, type Ⅱ) was microinjected into mouse 1-cell stage embryos at G2 phase, and MPF (M-phase promoting factor) activity as well as cyclin B concentration was measured. In PKI microinjected group, the PKA activity was low in the period between 27 - 29 hours after adding hCG, but MPF activity reached to a high value half an hour earlier and lasted longer than in control group. However, when high dosage PKI was microinjected, PKA activity kept low, MPF activity kept high and M/G1 arrest happened. Data from western blotting showed cyclin B concentration was high at metaphase in PKI injected group while cyclin B was degraded in the control group. These results indicated that inhibition of endogenous PKA in interphase increased MPF activity, while high dosage of PKI microinjection would induce M/ G1 arrest, most probably caused by cyclin B undegradation.

关 键 词：小鼠受精卵 早期发育 PKA M/G1期 蛋白激酶A M期促进因子 MPF 

分 类 号：Q492[生物学—生理学]