Ouabain facilitates cardiac differentiation of mouse embryonic stem cells through ERK1/2 pathway

Yee Ki Lee, Kwong Man Ng, Wing Hon Lai, Cornelia Man, Deborah Lieu, Chu Pak Lau, Hung Fat Tse, Chung Wah Siu

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Aim:To investigate the effects of the cardiotonic steroid, ouabain, on cardiac differentiation of murine embyronic stem cells (mESCs).Methods:Cardiac differentiation of murine ESCs was enhanced by standard hanging drop method in the presence of ouabain (20 μmol/L) for 7 d. The dissociated ES derived cardiomyocytes were examined by flow cytometry, RT-PCR and confocal calcium imaging.Results:Compared with control, mESCs treated with ouabain (20 μmol/L) yielded a significantly higher percentage of cardiomyocytes, and significantly increased expression of a panel of cardiac markers including Nkx 2.5, α-MHC, and Β-MHC. The α1 and 2- isoforms Na +/K +-ATPase, on which ouabain acted, were also increased in mESCs during differentiation. Among the three MAPKs involved in the cardiac hypertrophy pathway, ouabain enhanced ERK1/2 activation. Blockage of the Erk1/2 pathway by U0126 (10 μmol/L) inhibited cardiac differentiation while ouabain (20 μmol/L) rescued the effect. Interestingly, the expression of calcium handling proteins, including ryanodine receptor (RyR2) and sacroplasmic recticulum Ca 2+ ATPase (SERCA2a) was also upregulated in ouabain-treated mESCs. ESC-derived cardiomyocyes (CM) treated with ouabain appeared to have more mature calcium handling. As demonstrated by confocal Ca 2+ imaging, cardiomyocytes isolated from ouabain-treated mESCs exhibited higher maximum upstroke velocity (P<0.01) and maximum decay velocity (P0.05), as well as a higher amplitude of caffeine induced Ca 2+ transient (P<0.05), suggesting more mature sarcoplasmic reticulum (SR).Conclusion:Ouabain induces cardiac differentiation and maturation of mESC-derived cardiomyocytes via activation of Erk1/2 and more mature SR for calcium handling.

Original languageEnglish (US)
Pages (from-to)52-61
Number of pages10
JournalActa Pharmacologica Sinica
Volume32
Issue number1
DOIs
StatePublished - Jan 2011

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MAP Kinase Signaling System
Ouabain
Stem Cells
Cardiac Myocytes
Calcium
Ryanodine Receptor Calcium Release Channel
Sarcoplasmic Reticulum
Mouse Embryonic Stem Cells
Cardiac Glycosides
Cardiomegaly
Caffeine
Adenosine Triphosphatases
Cell Differentiation
Flow Cytometry
Protein Isoforms
Polymerase Chain Reaction

Keywords

  • cardiac differentiation
  • embryonic stem cells
  • ouabain

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)

Cite this

Ouabain facilitates cardiac differentiation of mouse embryonic stem cells through ERK1/2 pathway. / Lee, Yee Ki; Ng, Kwong Man; Lai, Wing Hon; Man, Cornelia; Lieu, Deborah; Lau, Chu Pak; Tse, Hung Fat; Siu, Chung Wah.

In: Acta Pharmacologica Sinica, Vol. 32, No. 1, 01.2011, p. 52-61.

Research output: Contribution to journalArticle

Lee, Yee Ki ; Ng, Kwong Man ; Lai, Wing Hon ; Man, Cornelia ; Lieu, Deborah ; Lau, Chu Pak ; Tse, Hung Fat ; Siu, Chung Wah. / Ouabain facilitates cardiac differentiation of mouse embryonic stem cells through ERK1/2 pathway. In: Acta Pharmacologica Sinica. 2011 ; Vol. 32, No. 1. pp. 52-61.
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AU - Ng, Kwong Man

AU - Lai, Wing Hon

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AU - Lieu, Deborah

AU - Lau, Chu Pak

AU - Tse, Hung Fat

AU - Siu, Chung Wah

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