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Reactive Oxygen Species as Messengers in Signaling Pathways Leading to Cardiovascular Differentiation
Author(s) -
Wartenberg Maria,
Sauer Heinrich
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a36-c
Subject(s) - vasculogenesis , mef2c , nadph oxidase , p38 mitogen activated protein kinases , angiogenesis , microbiology and biotechnology , reactive oxygen species , kinase , biology , mapk/erk pathway , vascular endothelial growth factor , chemistry , stem cell , cancer research , gene expression , biochemistry , progenitor cell , gene , vegf receptors
Human and mouse embryonic stem cells are able to differentiate into cardiomyocytes and blood vessels as well. In our studies mouse embryonic stem cells growing within multicellular tissues are subjected to reactive oxygen species (ROS), electromagnetic field (EM‐field) and mechanical forces which may initiate differentiation programs. Herein, we demonstrate that different noxe stimulated cardiovascular differentiation of mouse embryonic stem (ES) cells. Cardiomyocyte differentiation was evaluated by quantification of spontaneously contracting cardiac foci and RT‐PCR for early, intermediate and late marker genes. Angiogenesis/vasculogenesis was determined by quantification of PECAM‐1‐positive capillary areas and early and late endothelial marker genes. External ROS, EM‐field application and mechanical strain elevated intracellular reactive oxygen species (ROS) within minutes. This was followed by upregulation of NADPH oxidase subunits p22‐phox, p47‐phox, p67‐phox as well Nox‐4 and Nox‐1 mRNA within 24h. ROS stimulated the expression of vascular endothelial growth factor (VEGF) mRNA and protein as well as MEF2C and GATA‐4 mRNA, which are involved in cardiovascular development. Furthermore, phosphorylation of extracellular‐regulated kinase 1,2 (ERK1,2), p38, and c‐jun N‐terminal kinase (c‐Jun NH2‐terminal kinase (JNK)) was observed. Stimulation of cardiovascular commitment, VEGF, and MEF2C expression as well as MAPK activation were abolished in the presence of free radical scavengers, whereas GATA‐4 expression was increased. Cardiomyogenesis was significantly inhibited in the presence of the p38 inhibitor SB203580, the ERK1,2 inhibitor UO126, and the JNK inhibitor SP600125. Vasculogenesis/angiogenesis was blunted following inhibition of ERK1,2 and JNK, whereas p38 inhibition was ineffective. The data of the present study outline a role of ROS as signalling molecules in stimulated cardiovascular differentiation of ES cells, and point toward a microenvironment of elevated ROS required for signaling cascades initiating cardiovascular differentiation programs.