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Related Experiment Video

Updated: Feb 24, 2026

Analysis of Cardiomyocyte Development using Immunofluorescence in Embryonic Mouse Heart
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Nestin expression is dynamically regulated in cardiomyocytes during embryogenesis.

Vanessa Hertig1, Adrianna Matos-Nieves2, Vidu Garg2

  • 1Research Center, Montreal Heart Institute and Université de Montréal, Montréal, Québec, Canada.

Journal of Cellular Physiology
|August 24, 2017
PubMed
Summary

Nestin expression in developing hearts is negatively regulated by Tbx5 and Gata4. Dephosphorylated Yap-1 promotes cardiomyocyte cell cycle re-entry by upregulating nestin.

Keywords:
GATA4TBX5Yap-1cardiomyocytesembryogenesisnestin

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Area of Science:

  • Cardiovascular Development
  • Cell Biology
  • Gene Regulation

Background:

  • The role of transcriptional factors in regulating nestin expression in embryonic cardiomyocytes is largely unknown.
  • Nestin, an intermediate filament protein, is transiently expressed during cardiac embryogenesis.

Purpose of the Study:

  • To investigate the transcriptional regulation of nestin during cardiomyocyte development.
  • To elucidate the role of Yap-1 signaling in cardiomyocyte cell cycle re-entry.

Main Methods:

  • Immunohistochemistry in embryonic mouse hearts to detect nestin, Tbx5, Ki67, and Yap-1.
  • In vitro studies using neonatal rat ventricular cardiomyocytes treated with phorbol ester, MAPK inhibitors, and verteporfin.
  • Gene silencing using shRNA delivered via AAV9 to assess nestin's role in cell cycle progression.

Main Results:

  • Nestin expression in cardiomyocytes decreases during late embryogenesis and is inversely correlated with Tbx5 and Gata4 levels.
  • Tbx5+/- and Gata4+/- embryonic hearts show increased nestin expression, with prevalent Ki67 and Yap-1 in nestin-positive cardiomyocytes.
  • Dephosphorylated Yap-1 signaling upregulates nestin expression, promoting cell cycle re-entry and proliferation in neonatal cardiomyocytes.

Conclusions:

  • Tbx5 and Gata4 negatively regulate nestin expression during cardiac embryogenesis.
  • Dephosphorylated Yap-1 is crucial for cardiomyocyte cell cycle re-entry through nestin upregulation.
  • Yap-1 may play a similar role in Tbx5+/- and Gata4+/- mutant hearts.