Cytoskeleton-associated protein 5 and clathrin heavy chain binding regulates spindle assembly in mouse oocytes.
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.
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, The Research Center for Laboratory Animal Science, College of Life Science, Inner Mongolia University, Inner Mongolia, People's Republic of China.
Angeleem
A
Lu
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, The Research Center for Laboratory Animal Science, College of Life Science, Inner Mongolia University, Inner Mongolia, People's Republic of China.
Cheng-Jie
CJ
Zhou
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, The Research Center for Laboratory Animal Science, College of Life Science, Inner Mongolia University, Inner Mongolia, People's Republic of China.
Dong-Hui
DH
Wang
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, The Research Center for Laboratory Animal Science, College of Life Science, Inner Mongolia University, Inner Mongolia, People's Republic of China.
Zhe
Z
Han
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, The Research Center for Laboratory Animal Science, College of Life Science, Inner Mongolia University, Inner Mongolia, People's Republic of China.
Xiang-Wei
XW
Kong
Inner Mongolia People's Hospital, Hohhot, Inner Mongolia, People's Republic of China.
Yu-Zhen
YZ
Ma
Inner Mongolia People's Hospital, Hohhot, Inner Mongolia, People's Republic of China.
Zhi-Zhong
ZZ
Yun
The Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, The Research Center for Laboratory Animal Science, College of Life Science, Inner Mongolia University, Inner Mongolia, People's Republic of China.
Cheng-Guang
CG
Liang
D000818:Animals; D015153:Blotting, Western; D033922:Clathrin Heavy Chains / Q000378:metabolism*; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005455:Fluorescent Antibody Technique; D047468:Immunoprecipitation; D008540:Meiosis / Q000502:physiology; D051379:Mice; D018613:Microscopy, Confocal; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism*; D008941:Spindle Apparatus / Q000378:metabolism*
C587430:CKAP5 protein, mouse; D008869:Microtubule-Associated Proteins; D033922:Clathrin Heavy Chains
CKAP5; CLTC; Pathology Section; meiosis; oocyte; spindle