Role of TRPV1 in the Differentiation of Mouse Embryonic Stem Cells into Cardiomyocytes

PLoS One. 2015 Jul 24;10(7):e0133211. doi: 10.1371/journal.pone.0133211. eCollection 2015.

Abstract

Cytosolic Ca2+ ([Ca2+]i) is an important signal that regulates cardiomyocyte differentiation during cardiogenesis. TRPV1 is a Ca2+-permeable channel that is expressed in cardiomyocytes. In the present study, we utilized mouse embryonic stem cell-derived cardiomyocytes (mESC-CMs) as a model to investigate the functional role of TRPV1 in cardiomyocyte differentiation. Induction of embryonic stem cells into cardiomyocytes was achieved using embryoid body (EB)-based differentiation method. Quantitative PCRs showed an increased TRPV1 expression during the differentiation process. In [Ca2+]i measurement study, application of TRPV1 agonists, capsaicin and camphor, elicited a [Ca2+]i rise in mESC-CMs, the effect of which was abolished by TRPV1-shRNA. In functional study, treatment of EBs with TRPV1 antagonists (capsazepine and SB366791) and TRPV1-shRNA reduced the size of the EBs and decreased the percentage of spontaneously beating EBs. TRPV1 antagonists and TRPV1-shRNA also suppressed the expression of cardiomyocyte marker genes, including cardiac actin, c-TnT, c-TnI, and α-MHC. Taken together, this study demonstrated an important functional role of TRPV1 channels in the differentiation of mESCs into cardiomyocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Cell Differentiation / genetics*
  • Cells, Cultured
  • Gene Expression
  • Mice
  • Mouse Embryonic Stem Cells / cytology*
  • Mouse Embryonic Stem Cells / metabolism*
  • Myocytes, Cardiac / cytology*
  • Myocytes, Cardiac / metabolism*
  • RNA, Messenger / genetics
  • RNA, Small Interfering / genetics
  • TRPV Cation Channels / agonists
  • TRPV Cation Channels / antagonists & inhibitors
  • TRPV Cation Channels / genetics*

Substances

  • Biomarkers
  • RNA, Messenger
  • RNA, Small Interfering
  • TRPV Cation Channels
  • TRPV1 protein, mouse

Grants and funding

This work was supported by T13-706/11 (XY), AoE/M-05/12 (XY), CUHK2/CRF/11G (XY), CUHK478710 (XY), CUHK478011 (XY, YH), CUHK478413 (XY), CUHK478413 (XY), CUHK474711 (SYT), CUHK474412 (SYT), and CUHK474913 (SYT) from the Hong Kong RGC; National Science Foundation of China grant 31171100 (XY); Innovative Technology Fund of Innovation Technology Commission: Funding Support to Partner State Key Laboratories in Hong Kong (SYT). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.