This Article Full Text Full Text (PDF) Data Supplement All Versions of this Article: 110/17/2666    most recent 01.CIR.0000145608.80855.BCv1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Massion, P. B. Articles by Balligand, J.-L. Search for Related Content PubMed PubMed Citation Articles by Massion, P. B. Articles by Balligand, J.-L. Related Collections Contractile function

(Circulation. 2004;110:2666-2672.)
© 2004 American Heart Association, Inc.

Molecular Cardiology

Cardiomyocyte-Restricted Overexpression of Endothelial Nitric Oxide Synthase (NOS3) Attenuates ß-Adrenergic Stimulation and Reinforces Vagal Inhibition of Cardiac Contraction

Paul B. Massion, MD; Chantal Dessy, PhD; Fanny Desjardins, MS; Michel Pelat, PhD; Xavier Havaux, MS; Catharina Belge, MD; Pierre Moulin, MD; Yves Guiot, PhD; Olivier Feron, PhD; Stefan Janssens, MD, PhD; Jean-Luc Balligand, MD, PhD

From the Unit of Pharmacology and Therapeutics (P.B.M., C.D., F.D., M.P., C.B., O.F., J.-L.B.), FATH 5349, Unit of Cardiology (X.H.), Department of Medicine, Unit of Anatomopathology (P.M., Y.G.), Université Catholique de Louvain, Brussels, Belgium, and Division of Cardiology (S.J.), Katholieke Universiteit Leuven, Leuven, Belgium.

Correspondence to J.-L. Balligand, MD, PhD, Unit of Pharmacology and Therapeutics, FATH 5349, Department of Medicine, Université Catholique de Louvain (UCL), 53 ave Mounier, 1200 Brussels, Belgium. E-mail balligand{at}mint.ucl.ac.be

Received March 15, 2004; de novo received May 14, 2004; revision received June 10, 2004; accepted June 17, 2004.

Background— In the heart, nitric oxide synthases (NOS) modulate cardiac contraction in an isoform-specific manner, which is critically dependent on their cellular and subcellular localization. Defective NO production by NOS3 (endothelial NOS [eNOS]) in the failing heart may precipitate cardiac failure, which could be reversed by overexpression of NOS3 in the myocardium.

Methods and Results— We studied the influence of NOS3 in relation to its subcellular localization on the function of cardiomyocytes isolated from transgenic mice overexpressing NOS3 under the -myosin heavy chain promoter (NOS3-TG). Immunoblot analysis demonstrated moderate (5-fold) NOS3 overexpression in cardiomyocytes from NOS3-TG heterozygotes. Caveolar localization of transgenic eNOS was demonstrated by immunofluorescence, coimmunoprecipitation with caveolin-3, sucrose gradient fractionation, and immunogold staining revealed by electron microscopy. Compared with wild-type littermate, contractility of NOS3-TG cardiomyocytes analyzed by videomicroscopy revealed a lower incidence of spontaneous arrhythmic contractions (n=32, P<0.001); an attenuation of the ß-adrenergic positive inotropic response (isoproterenol, 10^–7 mol/L: 62.1±7.8% versus 90.8±8.0% of maximum Ca²⁺ response; n=10 to 17; P<0.05); a potentiation of the muscarinic negative chronotropic response (carbamylcholine, 3.10^–8 mol/L: –63.9±14% versus –27.7±5.6% of basal rate; n=8 to 10; P<0.05), confirmed by telemetry in vivo; and an attenuation of the accentuated antagonism of ß-adrenergically stimulated contraction (–14.6±1.5% versus –3.5±1.5; n=7 to 11; P<0.05). Cardiomyocyte NOS inhibition reversed all 4 effects (P<0.05).

Conclusions— Moderate overexpression of NOS3, targeted to caveolae in murine cardiomyocytes, potentiates the postsynaptic muscarinic response and attenuates the effect of high concentrations of catecholamines. Cardiomyocyte NOS3 may represent a promising therapeutic target to restore the sympathovagal balance and protect the heart against arrhythmia.

Key Words: contractility • nitric oxide synthase • arrhythmia • acetylcholine • catecholamines

This article has been cited by other articles:

				H. Wang, M. J. Kohr, D. G. Wheeler, and M. T. Ziolo Endothelial nitric oxide synthase decreases {beta}-adrenergic responsiveness via inhibition of the L-type Ca2+ current Am J Physiol Heart Circ Physiol, March 1, 2008; 294(3): H1473 - H1480. [Abstract] [Full Text] [PDF]

				J. Saliez, C. Bouzin, G. Rath, P. Ghisdal, F. Desjardins, R. Rezzani, L.F. Rodella, J. Vriens, B. Nilius, O. Feron, et al. Role of Caveolar Compartmentation in Endothelium-Derived Hyperpolarizing Factor-Mediated Relaxation: Ca2+ Signals and Gap Junction Function Are Regulated by Caveolin in Endothelial Cells Circulation, February 26, 2008; 117(8): 1065 - 1074. [Abstract] [Full Text] [PDF]

				K. Lemmens, K. Doggen, and G. W. De Keulenaer Role of Neuregulin-1/ErbB Signaling in Cardiovascular Physiology and Disease: Implications for Therapy of Heart Failure Circulation, August 21, 2007; 116(8): 954 - 960. [Abstract] [Full Text] [PDF]

				M. Seddon, A. M. Shah, and B. Casadei Cardiomyocytes as effectors of nitric oxide signalling Cardiovasc Res, July 15, 2007; 75(2): 315 - 326. [Abstract] [Full Text] [PDF]

				E. Takimoto, D. Belardi, C. G. Tocchetti, S. Vahebi, G. Cormaci, E. A. Ketner, A. L. Moens, H. C. Champion, and D. A. Kass Compartmentalization of Cardiac {beta}-Adrenergic Inotropy Modulation by Phosphodiesterase Type 5 Circulation, April 24, 2007; 115(16): 2159 - 2167. [Abstract] [Full Text] [PDF]

				F. Ichinose, E. S. Buys, T. G. Neilan, E. M. Furutani, J. G. Morgan, D. S. Jassal, A. R. Graveline, R. J. Searles, C. C. Lim, M. Kaneki, et al. Cardiomyocyte-Specific Overexpression of Nitric Oxide Synthase 3 Prevents Myocardial Dysfunction in Murine Models of Septic Shock Circ. Res., January 5, 2007; 100(1): 130 - 139. [Abstract] [Full Text] [PDF]

				B. Casadei The emerging role of neuronal nitric oxide synthase in the regulation of myocardial function Exp Physiol, November 1, 2006; 91(6): 943 - 955. [Abstract] [Full Text] [PDF]

				J. W. Elrod, J. J.M. Greer, N. S. Bryan, W. Langston, J. F. Szot, H. Gebregzlabher, S. Janssens, M. Feelisch, and D. J. Lefer Cardiomyocyte-Specific Overexpression of NO Synthase-3 Protects Against Myocardial Ischemia-Reperfusion Injury Arterioscler. Thromb. Vasc. Biol., July 1, 2006; 26(7): 1517 - 1523. [Abstract] [Full Text] [PDF]

				S. M. Davidson and M. R. Duchen Effects of NO on mitochondrial function in cardiomyocytes: Pathophysiological relevance Cardiovasc Res, July 1, 2006; 71(1): 10 - 21. [Abstract] [Full Text] [PDF]

				S. R. Martin, K. Emanuel, C. E. Sears, Y.-H. Zhang, and B. Casadei Are myocardial eNOS and nNOS involved in the {beta}-adrenergic and muscarinic regulation of inotropy? A systematic investigation Cardiovasc Res, April 1, 2006; 70(1): 97 - 106. [Abstract] [Full Text] [PDF]

				O. Feron and J.-L. Balligand Caveolins and the regulation of endothelial nitric oxide synthase in the heart Cardiovasc Res, March 1, 2006; 69(4): 788 - 797. [Abstract] [Full Text] [PDF]

				A. Godecke On the impact of NO-globin interactions in the cardiovascular system Cardiovasc Res, February 1, 2006; 69(2): 309 - 317. [Abstract] [Full Text] [PDF]

				J.-L. Balligand "La Donna e Mobile...": Is Cardiac Neuronal Nitric Oxide Synthase Such a Disconcerting Enzyme? Circulation, December 13, 2005; 112(24): 3668 - 3671. [Full Text] [PDF]

				F. U. Muller Increased eNOS expression as a compensatory mechanism reducing {beta}-adrenergic responsiveness? Cardiovasc Res, September 1, 2005; 67(4): 575 - 577. [Full Text] [PDF]

				E.J.F. Danson, Y.H. Zhang, C.E. Sears, A.R. Edwards, B. Casadei, and D.J. Paterson Disruption of inhibitory G-proteins mediates a reduction in atrial {beta}-adrenergic signaling by enhancing eNOS expression Cardiovasc Res, September 1, 2005; 67(4): 613 - 623. [Abstract] [Full Text] [PDF]

				M. Wehling-Henricks, M. C. Jordan, K. P. Roos, B. Deng, and J. G. Tidball Cardiomyopathy in dystrophin-deficient hearts is prevented by expression of a neuronal nitric oxide synthase transgene in the myocardium Hum. Mol. Genet., July 15, 2005; 14(14): 1921 - 1933. [Abstract] [Full Text] [PDF]

				J. O-Uchi, K. Komukai, Y. Kusakari, T. Obata, K. Hongo, H. Sasaki, and S. Kurihara {alpha}1-Adrenoceptor stimulation potentiates L-type Ca2+ current through Ca2+/calmodulin-dependent PK II (CaMKII) activation in rat ventricular myocytes PNAS, June 28, 2005; 102(26): 9400 - 9405. [Abstract] [Full Text] [PDF]

				C. BELGE, P. B. MASSION, M. PELAT, and J. L. BALLIGAND Nitric Oxide and the Heart: Update on New Paradigms Ann. N.Y. Acad. Sci., June 1, 2005; 1047(1): 173 - 182. [Abstract] [Full Text] [PDF]

				E. J. DANSON and D. J. PATERSON Cardiac Neurobiology of Nitric Oxide Synthases Ann. N.Y. Acad. Sci., June 1, 2005; 1047(1): 183 - 196. [Abstract] [Full Text] [PDF]

				G. F. Gonzales, F. A. Chung, S. Miranda, L. B. Valdez, T. Zaobornyj, J. Bustamante, and A. Boveris Heart mitochondrial nitric oxide synthase is upregulated in male rats exposed to high altitude (4,340 m) Am J Physiol Heart Circ Physiol, June 1, 2005; 288(6): H2568 - H2573. [Abstract] [Full Text] [PDF]