Cytoskeleton-associated protein 5 and clathrin heavy chain binding regulates spindle assembly in mouse oocytes.
Cytoskeleton-associated protein 5 and clathrin heavy chain binding regulates spindle assembly in mouse oocytes.
复制标题
细胞骨架相关蛋白 5 和网格蛋白重链结合调节小鼠卵母细胞中纺锤体的组装
DOI：
10.18632/oncotarget.15097
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发表时间：
2017-03-14
期刊：
影响因子：
--
通讯作者：
Liang CG
中科院分区：
文献类型：
--
作者：
Lu A;Zhou CJ;Wang DH;Han Z;Kong XW;Ma YZ;Yun ZZ;Liang CG
哺乳动物卵母细胞减数分裂成熟是早期胚胎发育的前提。卵母细胞成熟过程中有大量的微管相关蛋白参与。细胞凋亡相关蛋白5（CKAP 5）是XMAP 215家族的成员，在有丝分裂过程中调节微管动力学。然而，它在减数分裂中的作用尚未得到充分研究。本研究探讨了CKAP 5在小鼠卵母细胞减数分裂成熟和早期胚胎发育中的作用。Western印迹显示，CKAP 5的表达从GVBD开始增加，在中期维持在高水平，到1细胞晚期后下降。共聚焦显微镜显示，在间期（GV、PN或2-细胞期）没有CKAP 5的特异性积聚。然而，一旦细胞进入减数分裂或有丝分裂，CKAP 5定位于整个纺锤体。用微管蛋白干扰试剂诺考达唑（诱导MT解聚）或紫杉醇（防止MT解聚）处理卵母细胞不影响CKAP 5表达，但导致CKAP 5重排。此外，CKAP 5的敲低导致第一极体挤出失败、纺锤体组装的严重缺陷和染色体比对失败。CKAP 5的缺失也降低了早期胚胎发育潜力。此外，免疫共沉淀显示CKAP 5与网格蛋白重链1（CLTC）结合。这些结果表明，CKAP 5在卵母细胞成熟和早期胚胎发育中起重要作用，并且CKAP 5可能与CLTC共同作用于小鼠卵母细胞成熟。
Mammalian oocyte meiotic maturation is the precondition of early embryo development. Lots of microtubules (MT)-associated proteins participate in oocyte maturation process. Cytoskeleton-associated protein 5 (CKAP5) is a member of the XMAP215 family that regulates microtubule dynamics during mitosis. However, its role in meiosis has not been fully studied. Here, we investigated the function of CKAP5 in mouse oocyte meiotic maturation and early embryo development. Western blot showed that CKAP5 expression increased from GVBD, maintaining at high level at metaphase, and decreased after late 1-cell stage. Confocal microscopy showed there is no specific accumulation of CKAP5 at interphase (GV, PN or 2-cell stage). However, once cells enter into meiotic or mitotic division, CKAP5 was localized at the whole spindle apparatus. Treatment of oocytes with the tubulin-disturbing reagents nocodazole (induces MTs depolymerization) or taxol (prevents MTs depolymerization) did not affect CKAP5 expression but led to a rearrangement of CKAP5. Further, knock-down of CKAP5 resulted in a failure of first polar body extrusion, serious defects in spindle assembly, and failure of chromosome alignment. Loss of CKAP5 also decreased early embryo development potential. Furthermore, co-immunoprecipitation showed that CKAP5 bound to clathrin heavy chain 1 (CLTC). Taken together, our results demonstrate that CKAP5 is important in oocyte maturation and early embryo development, and CKAP5 might work together with CLTC in mouse oocyte maturation.