Journal
MICROSCOPY AND MICROANALYSIS
Volume 19, Issue 5, Pages 1364-1373Publisher
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S1431927613001943
Keywords
CLTC; spindle assembly; chromosome congression; tubulin; meiosis
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Funding
- National Basic Research Program of China [2011CB111500]
- National Natural Science Foundation of China [31160243]
- Open Major Basic Research Projects in Inner Mongolia
- Key Project of Chinese Ministry of Education [211029]
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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 beta-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.
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