Clathrin Heavy Chain 1 is Required for Spindle Assembly and Chromosome Congression in Mouse Oocytes
Clathrin Heavy Chain 1 is Required for Spindle Assembly and Chromosome Congression in Mouse Oocytes
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DOI：
10.1017/s1431927613001943
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发表时间：
2013-07
影响因子：
2.8
通讯作者：
Jie Zhao;Lu Wang;Hong-Xia Zhou;Li Liu;Angeleem Lu;Guangyao Li;H. Schatten;C. Liang
中科院分区：
文献类型：
--
作者：
Jie Zhao;Lu Wang;Hong-Xia Zhou;Li Liu;Angeleem Lu;Guangyao Li;H. Schatten;C. Liang
摘要网格蛋白重链1（CLTC）被认为是一种“兼职蛋白”，在分裂间期起膜运输作用，在有丝分裂期起稳定纺锤体纤维的作用。然而，其在减数分裂，特别是在哺乳动物卵母细胞成熟中的作用尚不清楚。本研究探讨了小鼠卵母细胞减数分裂成熟过程中CLTC的表达及其在纺锤体形成和染色体聚集中的作用。结果表明，CLTC在胚泡破裂（GVBD）后表达增加，并在M期达到高峰。免疫组化结果显示，CLTC分布在整个细胞质中的细胞周期依赖性的方式。在纺锤体的动态变化过程中，可观察到CLTC沿着β-微管蛋白（TUBB）的出现和消失。为了探讨CLTC与微管动力学的关系，用紫杉醇或诺考达唑处理中期卵母细胞。紫杉醇处理后，CLTC与TUBB共定位于膨大的纺锤体和细胞质中的星形细胞;诺考达唑处理后，CLTC与TUBB一起沿着分布于细胞质中。使用隐形siRNA破坏CLTC功能导致第一极体排出率降低和广泛的纺锤体形成和染色体聚集缺陷。这些结果表明，在小鼠卵母细胞成熟过程中，CLTC通过微管相关机制在纺锤体组装和染色体聚集中起重要作用。
Abstract Clathrin heavy chain 1 (CLTC) has been considered a “moonlighting protein” which acts in membrane trafficking during interphase and in stabilizing spindle fibers during mitosis. However, its roles in meiosis, especially in mammalian oocyte maturation, remain unclear. This study investigated CLTC expression and function in spindle formation and chromosome congression during mouse oocyte meiotic maturation. Our results showed that the expression level of CLTC increased after germinal vesicle breakdown (GVBD) and peaked in the M phase. Immunostaining results showed CLTC distribution throughout the cytoplasm in a cell cycle-dependent manner. Appearance and disappearance of CLTC along with β-tubulin (TUBB) could be observed during spindle dynamic changes. To explore the relationship between CLTC and microtubule dynamics, oocytes at metaphase were treated with taxol or nocodazole. CLTC colocalized with TUBB at the enlarged spindle and with cytoplasmic asters after taxol treatment; it disassembled and distributed into the cytoplasm along with TUBB after nocodazole treatment. Disruption of CLTC function using stealth siRNA caused a decreased first polar body extrusion rate and extensive spindle formation and chromosome congression defects. Taken together, these results show that CLTC plays an important role in spindle assembly and chromosome congression through a microtubule correlation mechanism during mouse oocyte maturation.