摘要
目的研究自动磷酸化Pak1在小鼠卵母细胞减数分裂过程中的亚细胞定位模式及其在纺锤体形成中的相关性。方法用Western blot检测Ser144位点自动磷酸化的Pak1(pPak1^(S144))的蛋白表达量的动态变化;免疫荧光染色追踪pPak1^(S144)的亚细胞定位模式及其与纺锤体和微管组织中心蛋白之间的时空关系。结果在小鼠卵母细胞内pPak1^(S144)持续稳定地表达于减数分裂各期,pPak1^(S144)与中心粒周围蛋白(pericentrin)和γ微管蛋白(γ-tubulin)在时空上呈现紧密的共定位关系并特异地定位于纺锤体两极。结论pPak1^(S144)是小鼠卵母细胞MTOC相关蛋白,提示其与减数分裂纺锤体形成相关。
Objective To investigate the subcellular localization of autophosphorylated Pakl in mouse oocytes during meiosis and its correlation with spindle formation. Methods Western blot procedure was employed to assay the dy- namic changes in protein expression of autophosphated Pakl at Ser144（pPaklS144） ; immunofluorescence staining was carried out to label pPaklSI144 subcellular distribution and its temporal-spatial relationship with spindle and microtubule organizing centers（MTOCs） proteins, Pericentrin and y-tubulin. Results pPakls144 was stably expressed during mei- otic progression in mouse oocytes; it was specifically co-localized with Pericentrin and y-tubulin and concentrated on spindle poles. Conclusions pPaklSI144 is MTOC-associated protein, implying a role in meiotic spindle formation.
出处
《基础医学与临床》
CSCD
北大核心
2014年第4期449-453,共5页
Basic and Clinical Medicine
基金
国家自然科学基金(31271253)
教育部博士点基金(20121107120019)
