Inhibition of cardiomyocytes differentiation of mouse embryonic stem cells by CD38/cADPR/Ca2+ signaling pathway.
Cyclic adenosine diphosphoribose (cADPR) is an endogenous Ca(2+) mobilizing messenger that is formed by ADP-ribosyl cyclases from nicotinamide adenine dinucleotide (NAD). The main ADP-ribosyl cyclase in mammals is CD38, a multi-functional enzyme and a type II membrane protein. Here we explored the role of CD38-cADPR-Ca(2+) in the cardiomyogenesis of mouse embryonic stem (ES) cells. We found that the mouse ES cells are responsive to cADPR and possess the key components of the cADPR signaling pathway. In vitro cardiomyocyte (CM) differentiation of mouse ES cells was initiated by embryoid body (EB) formation. Interestingly, beating cells appeared earlier and were more abundant in CD38 knockdown EBs than in control EBs. Real-time RT-PCR and Western blot analyses further showed that the expression of several cardiac markers, including GATA4, MEF2C, NKX2.5, and α-MLC, were increased markedly in CD38 knockdown EBs than those in control EBs. Similarly, FACS analysis showed that more cardiac Troponin T-positive CMs existed in CD38 knockdown or 8-Br-cADPR, a cADPR antagonist, treated EBs compared with that in control EBs. On the other hand, overexpression of CD38 in mouse ES cells significantly inhibited CM differentiation. Moreover, CD38 knockdown ES cell-derived CMs possess the functional properties characteristic of normal ES cell-derived CMs. Last, we showed that the CD38-cADPR pathway negatively modulated the FGF4-Erks1/2 cascade during CM differentiation of ES cells, and transiently inhibition of Erk1/2 blocked the enhanced effects of CD38 knockdown on the differentiation of CM from ES cells. Taken together, our data indicate that the CD38-cADPR-Ca(2+) signaling pathway antagonizes the CM differentiation of mouse ES cells.
Department of Physiology, The University of Hong Kong, Hong Kong, China.
Wen-Jie
WJ
Wei
Department of Physiology, The University of Hong Kong, Hong Kong, China.
Hai-Ying
HY
Sun
Department of Physiology, The University of Hong Kong, Hong Kong, China.
Kai Yiu
KY
Ting
State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
Li-He
LH
Zhang
Department of Physiology, The University of Hong Kong, Hong Kong, China.
Hon-Cheung
HC
Lee
Department of Physiology, The University of Hong Kong, Hong Kong, China.
Gui-Rong
GR
Li
Department of Physiology, The University of Hong Kong, Hong Kong, China. Electronic address: jyue@hku.hk.
Jianbo
J
Yue
D051997:ADP-ribosyl Cyclase 1 / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics; D020013:Calcium Signaling / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D036563:Cyclic ADP-Ribose / Q000235:genetics / Q000378:metabolism*; D058732:Embryoid Bodies / Q000166:cytology / Q000201:enzymology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000201:enzymology*; D051519:Fibroblast Growth Factor 4 / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology; D055785:Gene Knockdown Techniques; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000166:cytology / Q000201:enzymology*
D000943:Antigens, Differentiation; C496446:Fgf4 protein, mouse; D051519:Fibroblast Growth Factor 4; D008562:Membrane Glycoproteins; D009124:Muscle Proteins; D036563:Cyclic ADP-Ribose; C498155:Cd38 protein, mouse; D051997:ADP-ribosyl Cyclase 1