Cdc25B蛋白过表达可使小鼠胚胎突破2-细胞期阻滞  

CDC25B OVEREXPRESSION REVERSES TWO-CELL BLOCK IN MOUSE EMBRYOS

作　　者：崔城[1] 汤浩[1] 于秉治[2] 

机构地区：[1]中国医科大学基础医学院生理学教研室 [2]生物化学与分子生物学教研室,辽宁沈阳110001

出　　处：《中国应用生理学杂志》2008年第1期99-103,共5页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金资助项目(30570945)

摘　　要：目的:探讨Cdc25B蛋白过表达对小鼠2-细胞期胚胎发育的影响。方法:利用体外转录试剂盒将Cdc25B转录成mRNA,将mRNA显微注射入小鼠2-细胞期胚胎中,观察胚胎发育情况和卵裂率。用蛋白激酶活性测定方法和Western印迹分别检测Cdc25B蛋白过表达小鼠胚胎MPF的活性及Cdc2-Tyr15的磷酸化状态。结果:hCG后48 h,mRNA注射组有超过40%的2-细胞期胚胎分裂到4-细胞期而对照组仍停留在2-细胞期;激酶活性测定显示注射Cdc25B mRNA后,MPF的活性显著升高;Cdc2-Tyr15的磷酸化状态变化与激酶活性测定结果一致。结论:Cdc25B蛋白过表达可以激活有丝分裂促进因子(MPF),从而使小鼠2-细胞期胚胎突破2-细胞期阻滞,发育到4-细胞期。Aim： To explore the effect of CAc25B overexpreasion on the development of mouse two-cell embryos. Methods： The pBSK- CAc25B was in vitro transcribed into 5＇-capped mRNA for microinjection by using mMESSAGE mMACHINE kit. The CAc25B mRNA was microinjected into mouse embryos at two-cell stage in order to observe the embryonic development and cleavage rate. Using protein kinase activity assay and Western blot to detect the MPF activity as well as the phosphorylation status of CAc2-Tyrl5 in CAc25B overexpression group respectively. Results： The mouse embryos with CAc25B overexpreasion developed to the four-cell stage 48 h after the hCG injection with the percentage of cleavage over 40% compared with the embryos in control groups which still re- mained at the two-cell stage. Moreover, MPF activity increased significantly after Cdc25B mRNA injection. The phosphorylation status of CAc2-Tyrl5 was coincident with MPF activity. Conclusion： The results indicate that CAc25B overexpreasion in early mouse two-cell embryos reverses two-cell block and promotes their development into four-cell stage by activating MPF.

关 键 词：小鼠2-细胞期胚胎 CDC25B MPF 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]