© 1999 by European Society of Cardiology
Copyright © 1999, European Society of Cardiology
Nitric oxide synthase expression and role during cardiomyogenesis
aInstitute of Anatomy I, University of Cologne, Cologne, Germany
bInstitute of Neurophysiology, University of Cologne, Cologne, Germany
cDepartment of Neuroanatomy, University of Hamburg, Hamburg, Germany
* Corresponding author. Tel: +49-221-478-5202; fax: +49-221-478-6711 Addicks.Anatomie{at}uni-koeln.de
Objective: The aim of the present study was the investigation of the expression of NOS during cardiomyogenesis and its functional role. Design: The qualitative and quantitative expression of NOS isoforms during different stages of cardiac development was evaluated using immunocytochemistry and dot blots, respectively. The functional relevance of NOS expression during cardiomyogenesis was investigated using the in vitro ES cell-differentiation model and selective pharmacological agents. Results: On day 7.5 of embryonic development (E7.5) none of the NOS isoforms were expressed in the embryo, whereas the inducible (iNOS), as well as the endothelial (eNOS) isoforms were detected in the extraembryonic parts. In contrast, starting from E9.5 rat and murine embryos displayed prominent iNOS and eNOS expression. This was correlated with high expression of soluble guanylylcyclase (sGC) as well as high cyclic GMP (cGMP) content. During further development after E14.5 both, iNOS as well as eNOS, started to be downregulated and shortly prior to birth reduced staining for eNOS was found, whereas iNOS was hardly detectable. We further investigated whether NO plays a role for cardiomyogenesis, using in vitro ES cell-derived cardiomyocytes differentiating within embryoid bodies (EBs). The NOS expression pattern in these cells paralleled the one detected in vivo. We demonstrate that continuous incubation of EBs with the NOS inhibitors L-NMMA (2–10 mM) or L-NA (2–10 mM) for 4 to 9 days after plating resulted in a pronounced differentiation arrest of cardiomyocytes, whereas this effect could be reversed by coapplication of the NO-donor spermine-NONOate (10 µM). Conclusions: Both, iNOS and eNOS isoforms are prominently expressed during early stages of cardiomyogenesis. Around E14.5 NOS expression starts to decline. Moreover, the NO-generation is required for cardiomyogenesis since NOS inhibitors prevent the maturation of terminally differentiated cardiomyocytes using the ES cell system.
KEYWORDS Developmental biology; Gene expression; Nitric oxide; Signal transduction; Cell culture
1 Authors contributed equally to the manuscript.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  J.-L. Balligand, O. Feron, and C. Dessy eNOS Activation by Physical Forces: From Short-Term Regulation of Contraction to Chronic Remodeling of Cardiovascular Tissues Physiol Rev, April 1, 2009; 89(2): 481 - 534. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Mujoo, V. G. Sharin, N. S. Bryan, J. S. Krumenacker, C. Sloan, S. Parveen, L. E. Nikonoff, A. Y. Kots, and F. Murad Role of nitric oxide signaling components in differentiation of embryonic stem cells into myocardial cells PNAS, December 2, 2008; 105(48): 18924 - 18929. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Sasse, D. Malan, M. Fleischmann, W. Roell, E. Gustafsson, T. Bostani, Y. Fan, T. Kolbe, M. Breitbach, K. Addicks, et al. Perlecan is critical for heart stability Cardiovasc Res, December 1, 2008; 80(3): 435 - 444. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. D. Kumar, S.-K. Yong, S. T. Dheen, B.-H. Bay, and S. S.-W. Tay Cardiac Malformations Are Associated with Altered Expression of Vascular Endothelial Growth Factor and Endothelial Nitric Oxide Synthase Genes in Embryos of Diabetic Mice Experimental Biology and Medicine, November 1, 2008; 233(11): 1421 - 1432. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 I. A. Ionova, J. Vasquez-Vivar, J. Whitsett, A. Herrnreiter, M. Medhora, B. C. Cooley, and G. M. Pieper Deficient BH4 production via de novo and salvage pathways regulates NO responses to cytokines in adult cardiac myocytes Am J Physiol Heart Circ Physiol, November 1, 2008; 295(5): H2178 - H2187. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Loyer, P. Oliviero, T. Damy, E. Robidel, F. Marotte, C. Heymes, and J.-L. Samuel Effects of sex differences on constitutive nitric oxide synthase expression and activity in response to pressure overload in rats Am J Physiol Heart Circ Physiol, November 1, 2007; 293(5): H2650 - H2658. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Liu, Y. Jiang, H. Hao, K. Gupta, J. Xu, L. Chu, E. McFalls, J. Zweier, C. Verfaillie, and R. J. Bache Endothelial nitric oxide synthase is dynamically expressed during bone marrow stem cell differentiation into endothelial cells Am J Physiol Heart Circ Physiol, September 1, 2007; 293(3): H1760 - H1765. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Hammoud, F. Xiang, X. Lu, F. Brunner, K. Leco, and Q. Feng Endothelial nitric oxide synthase promotes neonatal cardiomyocyte proliferation by inhibiting tissue inhibitor of metalloproteinase-3 expression Cardiovasc Res, July 15, 2007; 75(2): 359 - 368. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Gassanov, M. Jankowski, B. Danalache, D. Wang, R. Grygorczyk, U. C. Hoppe, and J. Gutkowska Arginine Vasopressin-mediated Cardiac Differentiation: INSIGHTS INTO THE ROLE OF ITS RECEPTORS AND NITRIC OXIDE SIGNALING J. Biol. Chem., April 13, 2007; 282(15): 11255 - 11265. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Lepic, D. Burger, X. Lu, W. Song, and Q. Feng Lack of endothelial nitric oxide synthase decreases cardiomyocyte proliferation and delays cardiac maturation Am J Physiol Cell Physiol, December 1, 2006; 291(6): C1240 - C1246. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Ventura CAM and Cell Fate Targeting: Molecular and Energetic Insights into Cell Growth and Differentiation Evid. Based Complement. Altern. Med., September 1, 2005; 2(3): 277 - 283. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Madonna, P. Di Napoli, M. Massaro, A. Grilli, M. Felaco, A. De Caterina, D. Tang, R. De Caterina, and Y.-J. Geng Simvastatin Attenuates Expression of Cytokine-inducible Nitric-oxide Synthase in Embryonic Cardiac Myoblasts J. Biol. Chem., April 8, 2005; 280(14): 13503 - 13511. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. P. Jones, J. J. M. Greer, P. D. Ware, J. Yang, K. Walsh, and D. J. Lefer Deficiency of iNOS does not attenuate severe congestive heart failure in mice Am J Physiol Heart Circ Physiol, January 1, 2005; 288(1): H365 - H370. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Kanno, P. K. M. Kim, K. Sallam, J. Lei, T. R. Billiar, and L. L. Shears II Nitric oxide facilitates cardiomyogenesis in mouse embryonic stem cells PNAS, August 17, 2004; 101(33): 12277 - 12281. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. C. Heng, H. K. Haider, E. K.-W. Sim, T. Cao, and S. C. Ng Strategies for directing the differentiation of stem cells into the cardiomyogenic lineage in vitro Cardiovasc Res, April 1, 2004; 62(1): 34 - 42. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sachinidis, B. K. Fleischmann, E. Kolossov, M. Wartenberg, H. Sauer, and J. Hescheler Cardiac specific differentiation of mouse embryonic stem cells Cardiovasc Res, May 1, 2003; 58(2): 278 - 291. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. S. Bredt Nitric oxide signaling specificity -- the heart of the problem J. Cell Sci., January 1, 2003; 116(1): 9 - 15. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. L. Brutsaert Cardiac Endothelial-Myocardial Signaling: Its Role in Cardiac Growth, Contractile Performance, and Rhythmicity Physiol Rev, January 1, 2003; 83(1): 59 - 115. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G. F. Bronzwaer, C. Heymes, C. A. Visser, and W. J. Paulus Myocardial fibrosis blunts nitric oxide synthase-related preload reserve in human dilated cardiomyopathy Am J Physiol Heart Circ Physiol, January 1, 2003; 284(1): H10 - H16. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Q. Feng, W. Song, X. Lu, J. A. Hamilton, M. Lei, T. Peng, and S.-P. Yee Development of Heart Failure and Congenital Septal Defects in Mice Lacking Endothelial Nitric Oxide Synthase Circulation, August 13, 2002; 106(7): 873 - 879. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. G.F Bronzwaer, C. Zeitz, C. A Visser, and W. J Paulus Endomyocardial nitric oxide synthase and the hemodynamic phenotypes of human dilated cardiomyopathy and of athlete's heart Cardiovasc Res, August 1, 2002; 55(2): 270 - 278. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Dai, P. S. Brookes, V. M. Darley-Usmar, and P. G. Anderson Bioenergetics in cardiac hypertrophy: mitochondrial respiration as a pathological target of NO{middle dot} Am J Physiol Heart Circ Physiol, December 1, 2001; 281(6): H2261 - H2269. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Peunova, V. Scheinker, H. Cline, and G. Enikolopov Nitric Oxide Is an Essential Negative Regulator of Cell Proliferation in Xenopus Brain J. Neurosci., November 15, 2001; 21(22): 8809 - 8818. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Munch, B. Bolck, A. Sugaru, K. Brixius, W. Bloch, and R. H.G. Schwinger Increased Expression of Isoform 1 of the Sarcoplasmic Reticulum Ca2+-Release Channel in Failing Human Heart Circulation, June 5, 2001; 103(22): 2739 - 2744. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. C. Lee, Y. D. Zhao, D. W. Courtman, and D. J. Stewart Abnormal Aortic Valve Development in Mice Lacking Endothelial Nitric Oxide Synthase Circulation, May 23, 2000; 101(20): 2345 - 2348. [Abstract] [Full Text] [PDF]
|
|