吉林大学学报(医学版)

• 基础研究 •    下一篇

蛋白激酶Wee1B在小鼠1-细胞期受精卵中的表达及其定位

刘超1,2,肖建英3,任丽莉3,刘洋3,马浚仁3,于秉治1   

  1. (1.中国医科大学生物化学与分子生物学教研室,辽宁 沈阳 110001;2.辽宁医学院发育生物学教研室,辽宁 锦州 121001;3.辽宁医学院生物化学与分子生物学教研室,辽宁 锦州 121001)
  • 收稿日期:2013-04-09 出版日期:2013-09-28 发布日期:2013-09-28
  • 通讯作者: 于秉治 E-mail:(Tel:024-23261253,E-mail:ybzbiochem@yeah.net)
  • 作者简介:刘 超(1978-),女,辽宁省铁岭市人,副教授,医学博士,主要从事分子生殖和分子肿瘤学的研究。
  • 基金资助:

    国家自然科学基金资助课题(81270654,81270698);辽宁省教育厅科研基金资助课题(L2012303)

Expression and localization of protein kinase Wee1B in mouse one-cell-stage embryos

LIU Chao1,2,XIAO Jian-ying3,REN Li-li3,LIU Yang3,MA Jun-ren3,YU Bing-zhi1   

  1. (1.Department of Biochemisty and Molecular Biology,China Medical University,Shenyang 110001,China;2.Department of Developmental Biology,Liaoning Medical University,Jinzhou 121001,China;3.Department of Biochemisty and Molecular Biology,Liaoning Medical University,Jinzhou 121001,China)
  • Received:2013-04-09 Online:2013-09-28 Published:2013-09-28

摘要:

目的:检测小鼠1-细胞期受精卵各个细胞周期中蛋白激酶Wee1B的表达水平及其定位水平,初步阐明Wee1B对哺乳动物受精卵早期发育调控的机制。方法:超排卵获得小鼠1-细胞期受精卵,收集卵细胞后用RT-PCR和Western blotting方法检测Wee1B的mRNA和蛋白表达水平;放射自显影测定Wee1B活性;间接免疫荧光观察Wee1B在各个细胞时期的定位。结果:小鼠1-细胞期受精卵G1、S、G2和M期Wee1B mRNA相对表达量比较差异均无统计学意义(P>0.05);S、G2和M期Wee1B蛋白表达量与G1期比较差异有统计学意义(P<0.01);与G1期比较,S、G2期Wee1B活性逐渐增高(P>0.05, P<0.01), M期Wee1B活性迅速下降(P<0.01)。G1期和S期细胞Wee1B蛋白偶联的红色荧光信号主要集中分布于细胞核,但在G2期细胞红色荧光信号弥散分布于细胞浆,在M末期和2-细胞期可见红色荧光信号又重新分布于细胞核,细胞浆内荧光减弱。结论:蛋白激酶Wee1B在小鼠1-细胞期受精卵各个细胞周期动态表达,Wee1B蛋白存在核浆转位,Wee1B的定位变化可能调控小鼠受精卵有丝分裂。

关键词: Wee1B, Wee1蛋白, 小鼠, 合子, 蛋白表达, 蛋白定位

Abstract:

Abstract:Objective
To observe the expression and localization of protein kinase Wee1B in mouse one-cell-stage embryos, and to pay foundation for further study on the mechanism of the early development of mouse embryos. Methods One-cell-stage  mouse embryos were collected after superovulation,and the mRNA and protein expression levels of Wee1B were examined by RT-PCR and Western blotting,respectively.The Wee1B activity in mouse embryos was detected by autoradiography and the localization of Wee1B in each cell cycle was observed by indirect immunofluorescence.Results There were no significant differences in the expression levels of Wee1B mRNA between G1, S, G2 and M phases (P>0.05); but there were significant differences in the expression levels of Wee1B protein between S, G2 , M phases and G1 phase (P<0.01).Compared with G1 phase, the activities of Wee1B in G2  phase and S phase were increased (P>0.05, P<0.01),but the activity of Wee1B in M  phase was decreased (P<0.01). Wee1B  distributed in the nucleus at the G1 and S phases and translocated to the cytoplasm at the G2/M transition in one-cell-stage embryos.Conclusion The protein and activity of Wee1B are dynamically expressed in one-cell-stage embryos and the nucleocytoplasmic shuttling of Wee1B may be the mechanism of early development of mouse embryos. 

Key words: Wee1B;Wee1 protein;mouse zygote;protein expression, protein localization

中图分类号: 

  • Q132