Journal article

Publication year: 2015

Pages: 23-33

Journal: International Journal of Cardiology

Volume number: 193

ISSN: 0167-5273

eISSN: 1874-1754

DOI Link: https://doi.org/10.1016/j.ijcard.2015.05.019

Publisher: Elsevier

Abstract: 

Background: Cardiotrophin-1 (CT-1) controls cardiomyogenesis of mouse embryonic stem (ES) cells.

Objectives: To investigate the signaling pathway underlying the action of CT-1 on cardiac cell differentiation.

Methods: Protein expression was analyzed by western blot technique and cardiac areas by immunohistochemistry. Calcium, reactive oxygen species (ROS) and nitric oxide (NO) were assessed by microfluorometry using fluo-4, H2DCF, and DAF-2DA, respectively. Gene inactivation of CT-1 was achieved by siRNA technology.

Results: CT-1 as well as its receptor gp 130 were transiently upregulated during differentiation of ES cells. Exogenous CT-1 enhanced cardiomyogenesis, increased the cardiac transcription factors MEF2c, Nkx-2.5, TEAD3 and GATA4, the cardiac proteins alpha-actinin, MLC2a, MYH7, MLC1a, MLC2v and HCN4 as well as vascular endothelial growth factor (VEGF), platelet-derived growth factor-BB (PDGF-BB), fibroblast growth factor-2 (FGF-2) and atrial natriuretic peptide (ANP). CT-1 downregulation by small interfering RNA (siRNA) inhibited cardiomyogenesis and decreased VEGF, PDGF-BB, FGF-2 and ANP expression. CT-1 raised intracellular calcium which was abolished by the intracellular calcium chelator BAPTA, AM and thapsigargin. Moreover, CT-1 treatment increased ROS, followed by NO generation and NOS3 activation. During ES cell differentiation CT-1 was translocated to the cell nucleus. Exogenous CT-1 induced nuclear translocation of endogenous CT-1, which was inhibited by BAPTA, the NOS inhibitor L-NG-Nitroarginine methyl ester (L-NAME), the radical scavenger N-(2-mercaptopropionyl)-glycine (NMPG) as well as the janus kinase 2 (JAK2) inhibitor AG490 and the PI3 kinase (PI3K) inhibitor LY294002.

Conclusions: Nuclear translocation of CT-1 regulates cardiomyogenesis of ES cells and involves calcium, NO, ROS as well as CT-1 regulated signaling pathways. (C) 2015 Published by Elsevier Ireland Ltd.

Harvard Citation style: Mascheck, L., Sharifpanah, F., Tsang, S., Wartenberg, M. and Sauer, H. (2015) Stimulation of cardiomyogenesis from mouse embryonic stem cells by nuclear translocation of cardiotrophin-1, International Journal of Cardiology, 193, pp. 23-33. https://doi.org/10.1016/j.ijcard.2015.05.019

APA Citation style: Mascheck, L., Sharifpanah, F., Tsang, S., Wartenberg, M., & Sauer, H. (2015). Stimulation of cardiomyogenesis from mouse embryonic stem cells by nuclear translocation of cardiotrophin-1. International Journal of Cardiology. 193, 23-33. https://doi.org/10.1016/j.ijcard.2015.05.019