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PLK1 regulates spindle formation kinetics and APC/C activation in mouse zygote

Published online by Cambridge University Press:  15 July 2015

Vladimir Baran*
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Soltesovej 4, 040 01 Kosice, Slovakia
Adela Brzakova
Affiliation:
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov, Czech Republic.
Pavol Rehak
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Kosice, Slovakia.
Veronika Kovarikova
Affiliation:
Institute of Animal Physiology, Slovak Academy of Sciences, Kosice, Slovakia.
Petr Solc
Affiliation:
Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Libechov, Czech Republic.
*
All correspondence to: Vladimir Baran. Institute of Animal Physiology, Slovak Academy of Sciences, Soltesovej 4, 040 01 Kosice, Slovakia. E-mail: baran@saske.sk

Summary

Polo-like kinase 1 (PLK1) is involved in essential events of cell cycle including mitosis in which it participates in centrosomal microtubule nucleation, spindle bipolarity establishment and cytokinesis. Although PLK1 function has been studied in cycling cancer cells, only limited data are known about its role in the first mitosis of mammalian zygotes. During the 1-cell stage of mouse embryo development, the acentriolar spindle is formed and the shift from acentriolar to centrosomal spindle formation progresses gradually throughout the preimplantation stage, thus providing a unique possibility to study acentriolar spindle formation. We have shown previously that PLK1 activity is not essential for entry into first mitosis, but is required for correct spindle formation and anaphase onset in 1-cell mouse embryos. In the present study, we extend this knowledge by employing quantitative confocal live cell imaging to determine spindle formation kinetics in the absence of PLK1 activity and answer the question whether metaphase arrest at PLK1-inhibited embryos is associated with low anaphase-promoting complex/cyclosome (APC/C) activity and consequently high securin level. We have shown that inhibition of PLK1 activity induces a delay in onset of acentriolar spindle formation during first mitosis. Although these PLK1-inhibited 1-cell embryos were finally able to form a bipolar spindle, not all chromosomes were aligned at the metaphase equator. PLK1-inhibited embryos were arrested in metaphase without any sign of APC/C activation with high securin levels. Our results document that PLK1 controls the onset of spindle assembly and spindle formation, and is essential for APC/C activation before anaphase onset in mouse zygotes.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2015 

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