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ROCK inhibition prevents early mouse embryo development

Abstract

ROCK is a Rho-GTPase effector that is important for actin assembly and is involved in various cellular functions, including cell contraction, migration, motility, and tumor cell invasion. In this study, we investigated ROCK expression and function during early mouse embryo development. Inhibiting ROCK by Y-27632 treatment at the zygote stage resulted in first cleavage failure, and most embryos failed to develop to the 8-cell stage. When adding Y-27632 at the 8-cell stage, embryos failed to undergo compaction and could not develop into blastocysts. In addition, fluorescence staining intensity analysis indicated that actin expression at blastomere membranes was significantly reduced. After ROCK inhibition, two or more nuclei were observed in a cell, which indicated possible cytokinesis failure. Moreover, after ROCK inhibition with Y-27632, the phosphorylation levels of LIMK1/2, a downstream molecule of ROCK, were decreased at blastomere membranes. Thus, our results showed conserved roles for ROCK in this mammalian embryo model and indicated that a ROCK-LIMK1/2-actin pathway might regulate cleavage and blastocyst formation during early mouse embryo development.

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Acknowledgments

This study was supported by the Natural Science Foundation of Jiangsu Province (BK20130671), Fundamental Research Funds for the Central Universities (KYRC201202), China, and the Biogreen 21 Program (PJ009594 & PJ00909801), RDA, Republic of Korea.

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The authors have no conflict of interest to disclose.

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Correspondence to Shao-Chen Sun.

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Duan, X., Chen, KL., Zhang, Y. et al. ROCK inhibition prevents early mouse embryo development. Histochem Cell Biol 142, 227–233 (2014). https://doi.org/10.1007/s00418-014-1201-6

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  • DOI: https://doi.org/10.1007/s00418-014-1201-6

Keywords

  • Embryo
  • Actin
  • ROCK
  • p-LIMK1/2
  • Blastocyst