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Related Experiment Video

Updated: Jun 6, 2026

Methods for Precisely Localized Transfer of Cells or DNA into Early Postimplantation Mouse Embryos
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Methods for Precisely Localized Transfer of Cells or DNA into Early Postimplantation Mouse Embryos

Published on: December 25, 2015

[Cdc25B overexpression reverses two-cell block in mouse embryos].

Cheng Cui1, Hao Tang, Bingzhi Yu

  • 1Department of Physiology, China Medical University, Shenyang 110001, China.

Zhongguo Ying Yong Sheng Li Xue Za Zhi = Zhongguo Yingyong Shenglixue Zazhi = Chinese Journal of Applied Physiology
|December 15, 2010
PubMed
Summary

Overexpressing Cdc25B in mouse embryos overcomes developmental arrest at the two-cell stage. This promotes progression to the four-cell stage by activating maturation-promoting factor (MPF).

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Area of Science:

  • Developmental Biology
  • Cell Cycle Regulation
  • Molecular Embryology

Background:

  • The transition from the two-cell to the four-cell stage in mouse embryos is a critical developmental checkpoint.
  • The two-cell block is a phenomenon where embryos arrest at this stage, hindering development.
  • Cdc25B is a key phosphatase involved in cell cycle progression, particularly in regulating MPF activity.

Purpose of the Study:

  • To investigate the impact of Cdc25B overexpression on the developmental progression of mouse embryos.
  • To determine if Cdc25B can overcome the natural two-cell block in early mouse development.

Main Methods:

  • In vitro transcription of Cdc25B mRNA using the mMESSAGE mMACHINE kit.
  • Microinjection of Cdc25B mRNA into mouse embryos at the two-cell stage.

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  • Assessment of embryonic development, cleavage rates, and protein kinase activity (MPF).
  • Western blot analysis to detect the phosphorylation status of Cdc2-Tyr15.

    • Main Results:

    • Cdc25B overexpression significantly increased the cleavage rate, with over 40% of embryos reaching the four-cell stage within 48 hours.
    • Control embryos remained arrested at the two-cell stage.
    • Maturation-promoting factor (MPF) activity was significantly elevated in Cdc25B-overexpressing embryos.
    • The phosphorylation status of Cdc2-Tyr15 correlated directly with increased MPF activity.

    Conclusions:

    • Cdc25B overexpression effectively reverses the two-cell block in early mouse embryos.
    • This reversal promotes embryonic development to the four-cell stage.
    • The mechanism involves the activation of MPF, facilitated by Cdc25B.