基础医学与临床 ›› 2018, Vol. 38 ›› Issue (3): 344-349.

• 研究论文 • 上一篇    下一篇

小鼠卵母细胞减数分裂过程中LIMK1参与调控Aurora-A在纺锤体的定位

姜秀颖,王思敏,周晴,翁静,马伟   

  1. 首都医科大学
  • 收稿日期:2017-08-23 修回日期:2017-12-22 出版日期:2018-03-05 发布日期:2018-02-27
  • 通讯作者: 马伟 E-mail:mawei1026@ccmu.edu.cn
  • 基金资助:
    国家自然科学基金

LIMK1 takes part in the regulation of Aurora-A-localization on spindle during mouse oocyte meiosis

  • Received:2017-08-23 Revised:2017-12-22 Online:2018-03-05 Published:2018-02-27

摘要: 目的 研究小鼠卵母细胞减数分裂进程中活化的LIMK1(pLIMK1Thr508)与Aurora-A亚细胞分布的相关性,LIMK1活性变化对Aurora-A和纺锤体组装的影响。方法 收集21日龄CB6F1小鼠卵母细胞,分别体外培养至生发泡期(GV)、生发泡破裂期(GVBD)、第1次减数分裂中期(MI)、第1次减数分裂后期(AI)和末期(Tel I)以及第2次减数分裂中期(MII),固定并利用免疫荧光染色法分析减数分裂进程中pLIMK1Thr508的亚细胞定位模式及其与纺锤体组装调控因子Aurora-A的时空相关性;利用抑制剂BMS-3处理MI期细胞,结合免疫荧光染色分析LIMK1活性缺失对于Aurora-A空间分布和纺锤体形成的影响。结果 在小鼠卵母细胞减数分裂前期(prophase)(即GV期),pLIMK1Thr508信号微弱并主要聚集于生发泡,此时Aurora-A在胞内没有特殊聚集;在临近减数分裂重启时,pLIMK1Thr508离开生发泡,Aurora-A信号出现,两者呈高度致密的点状聚集并紧密重合;在生发泡完全破裂后,pLIMK1Thr508和Aurora-A又呈多个点片状聚集在凝集的染色体周围;在MI期和MII期,pLIMK1Thr508与Aurora-A共同定位于纺锤体两极;在从AI期到Tel I期进程中pLIMK1Thr508离开纺锤体两极,聚集在收缩环部位,Aurora-A在纺锤体两极有微弱聚集。LIMK1抑制剂BMS-3能够破坏Aurora-A在纺锤体两极的聚集,并影响纺锤体结构。结论 pLIMK1Thr508在卵母细胞减数分裂中是一种MTOC相关蛋白,可能通过调控Aurora-A而参与纺锤体结构的形成和维持。

关键词: 关键词:pLIMK1Thr508, Aurora-A, 卵母细胞, 减数分裂, 纺锤体

Abstract: Objective To investigate the sub-cellular distribution correlation between activated LIMK1 (pLIMK1Thr508) and Aurora-A in mouse oocyte meiosis, and changes in Aurora-A location and spindle structure in condition of LIMK1 inhibition. Methods Immunofluorescence staining was employed to detect the sub-cellular localization of pLIMK1Thr508 and its spatial-temporal correlation with spindle organizing regulator Aurora-A in mouse oocyte meiosis; BMS-3, the specific inhibitor to LIMK1 activity, was applied to analyze the effects of LIMK1 inhibition on Aurora-A distribution and spindle formation. Results At meiotic prophase, pLIMK1Thr508 was weakly detected and concentrated in the germinal vesicle (GV) in oocytes, with no signal of Aurora-A across the cytoplasm and nuclear area; as meiotic assumption approaching, pLIMK1Thr508 left nuclear, aggregating as a single dense dote in the vicinity of nuclear, and being co-localized with the emerging Aurora-A; After germinal vesicle broke down (GVBD), pLIMK1Thr508 and Aurora-A remained overlapped and concentrated as multi foci around the condensed chromosomes; at metaphase I (MI) and metaphase II (MII), pLIMK1Thr508 was co-localized with Aurora-A on spindle poles; During anaphase I (AI) to telophase I (Tel I) progression, pLIMK1Thr508 was detached from spindle poles and mainly concentrated on the cleavage furrow, while Aurora-A loosely congressed on spindle. In addition, LIMK1 inhibition with BMS-3 destroyed Aurora-A polar location and spindle formation. Conclusions pLIMK1Thr508 is a microtubule organizing center (MTOC)-associated protein, may participate in spindle assembly and maintenance through regulating Aurora-A in mouse oocytes during meiotic progression.

Key words: Key words: pLIMK1Thr508, Aurora-A, oocytes, meiosis, spindle

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