摘要
目的:检测小鼠1-细胞期受精卵各细胞周期中细胞分裂周期蛋白14B(CDC14B)的表达水平及其定位,初步阐明CDC14B对哺乳动物受精卵早期发育调控的机制。方法:通过超排卵技术分别收集小鼠1-细胞期受精卵,采用RT-PCR和Western blotting法检测小鼠不同细胞周期1-细胞期受精卵中CDC14B mRNA和蛋白表达水平;采用细胞免疫荧光法观察小鼠不同细胞周期1-细胞期受精卵中CDC14B和β微管蛋白(β-tubulin)的共定位情况。结果:与G 1期比较,小鼠S、G 2和M期1-细胞期受精卵中CDC14B mRNA和蛋白表达水平明显升高(P<0.05或P<0.01)。间接免疫荧光实验,CDC14B(红色荧光)和β-tubulin(绿色荧光)在G 1期和S期共定位于小鼠受精卵皮质;在G 2早期,CDC14B和β-tubulin共定位于皮质和细胞质;在G 2晚期,部分CDC14B入核;在M期,CDC14B和β-tubulin共定位于纺锤体。结论:CDC14B在小鼠不同细胞周期1-细胞期受精卵中动态表达,CDC14B存在核浆纺锤体转位,CDC14B的定位变化可能调控纺锤体的功能。
Objective:To observe the expression and localization of cell division cycle 14B(CDC14B)in each cell cycle of the mouse one-cell fertilized eggs,and to elucidate preliminarily the mechanism of CDC14B regulating the early development of mammalian fertilized eggs.Methods:The superovulation technique was used to collect the mouse one-cell fertilized eggs.The expression levels of CDC14B mRNA and protein in the mouse one-cell stage fertilized eggs in different cell cycles were detected by RT-PCR and Western blotting methods,respectively.Co-localization of CDC14B andβ-tubulin in the mouse one-cell fertilized eggs in different cell cycles was observed by cell immunofluorescence method.Results:Compared with G 1 phase,the expression levels of CDC14B mRNA and protein in the mouse one-cell fertilized eggs in S,G 2 and M phases were significantly increased(P<0.05 or P<0.01).The cellular immunofluorescence assay results showed that CDC14B(red fluorescence)andβ-tubulin(green fluorescence)were co-located in the cortex of fertilized mouse eggs in G 1 phase and S phase,in the early G 2 phase,CDC14B andβ-tubulin co-localized in the cortex and cytoplasm;in the late G 2 phase,part of CDC14B entered the nucleus;in the M phase,CDC14B andβ-tubulin co-localized in the spindle.Conclusion:CDC14B dynamically expresses in each cell cycle of the mouse one-cell fertilized eggs.CDC14B has nuclear plasma spindle translocation,and the localization changes of CDC14B may regulate the spindle function.
作者
刘岚清
李瑞林
周晓敏
孟峻
LIU Lanqing;LI Ruilin;ZHOU Xiaomin;MENG Jun(Graduate College,Inner Mongolia Medical University,Hohhot 010059,China;Department of Laboratory,Affiliated Hospital,Inner Mongolia Medical University,Hohhot010050,China)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2020年第4期675-679,共5页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金资助课题(81360109,81660267)
内蒙古自治区科技厅自然科学基金资助课题(2013MS1163)。
