© 1996 American Heart Association, Inc.
Articles |
Nitric Oxide Synthase (NOS3)–Mediated Cholinergic Modulation of Ca2+ Current in Adult Rabbit Atrioventricular Nodal Cells
From the Cardiovascular Division, Department of Medicine (X.H., J.-L.B., R.A.K., T.W.S.) and Department of Pathology (L.K.), Brigham and Women's Hospital, Harvard Medical School, and Harvard School of Public Health, Boston, Mass.
Correspondence to Dr X. Han, Cardiovascular Division, Brigham and Women's Hospital, Harvard Medical School, 75 Francis St, Boston, MA 02115. E-mail xhan@bics.bwh.harvard.edu.
Abstract We examined the role of endogenous NO in the autonomic regulation of atrioventricular (AV) nodal function by studying spontaneous action potentials (SAPs) and L-type Ca2+ current (ICa-L) in isolated single AV nodal cells from adult rabbit hearts. Both the perforated and the membrane-ruptured patch-clamp techniques in the whole-cell configuration were used under conditions known to alter NO production. Three NO donors, 3-morpholinosydnonimine (SIN-1, 0.1 mmol/L), S-nitroso-acetylcysteine (0.1 mmol/L), and sodium nitroprusside (0.1 mmol/L), suppressed the ß-adrenergic agonist isoproterenol (ISO, 1 µmol/L)–stimulated increase in ICa-L. SIN-1 also decreased the frequency and amplitude of SAPs. In cells in which ICa-L had been previously attenuated by the muscarinic agonist carbamylcholine (CCh, 1 µmol/L), SIN-1 had no additive effect. CCh activated an acetylcholine-sensitive outward K+ current (IK(ACh)) in AV nodal cells, in addition to the ICa-L inhibition. Intracellular dialysis with the NO synthase inhibitor N-monomethyl-L-arginine (L-NMMA, 0.5 mmol/L) blocked CCh-induced, but not SIN-1–induced, ICa-L attenuation. However, intracellular dialysis with methylene blue (20 µmol/L), which inhibits NO-mediated activation of guanylyl cyclase and cGMP production, blocked the effects of both CCh and SIN-1 on ICa-L. In these cells, neither L-NMMA nor methylene blue affected the CCh-activated IK(ACh). Direct application of cGMP (10 µmol/L) via internal dialysis significantly inhibited ISO-stimulated ICa-L. In AV nodal cells internally perfused with either a nonhydrolyzable cAMP analogue, 8-Br-cAMP (0.5 mmol/L), or a high concentration of cAMP (0.5 mmol/L), CCh did not inhibit ICa-L but still activated IK(ACh). CCh-induced ICa-L attenuation could be abolished or quickly reversed by the nonselective phosphodiesterase (PDE) inhibitor 3-isobutyl-1-methylxanthine (20 µmol/L). However, CCh still significantly suppressed ISO-stimulated ICa-L after the cGMP-inhibited PDE isozyme (PDE3) had been selectively inhibited by milrinone (5 µmol/L). Immunohistochemical staining identified the presence of the endothelial constitutive NO synthase (ecNOS or NOS3) in both single AV nodal cells in vitro and in cryostat sections of AV nodal tissue in situ. These results demonstrate that endogenous NO is involved in the muscarinic cholinergic attenuation of ICa-L in AV nodal cells; the mechanism likely involves the cGMP-stimulated PDE.
Key Words: nitric oxide • atrioventricular node • whole-cell patch clamp • isoproterenol • carbamylcholine
This article has been cited by other articles:
 |
|
 |
|
  C. P. Ye, S. Z. Duan, D. S. Milstone, and R. M. Mortensen Go controls the hyperpolarization-activated current in embryonic stem cell-derived cardiocytes Am J Physiol Heart Circ Physiol, February 1, 2008; 294(2): H979 - H985. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F. Vandeput, S. L. Wolda, J. Krall, R. Hambleton, L. Uher, K. N. McCaw, P. B. Radwanski, V. Florio, and M. A. Movsesian Cyclic Nucleotide Phosphodiesterase PDE1C1 in Human Cardiac Myocytes J. Biol. Chem., November 9, 2007; 282(45): 32749 - 32757. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Zaccolo and M. A. Movsesian cAMP and cGMP Signaling Cross-Talk: Role of Phosphodiesterases and Implications for Cardiac Pathophysiology Circ. Res., June 8, 2007; 100(11): 1569 - 1578. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Mongillo, C. G. Tocchetti, A. Terrin, V. Lissandron, Y.-F. Cheung, W. R. Dostmann, T. Pozzan, D. A. Kass, N. Paolocci, M. D. Houslay, et al. Compartmentalized Phosphodiesterase-2 Activity Blunts {beta}-Adrenergic Cardiac Inotropy via an NO/cGMP-Dependent Pathway Circ. Res., February 3, 2006; 98(2): 226 - 234. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Z. Zhu, C. L. Chong, S. C. Chuah, S. H. Huang, H. S. Nai, H. T. Tong, M. Whiteman, and P. K. Moore Cardioprotective effects of nitroparacetamol and paracetamol in acute phase of myocardial infarction in experimental rats Am J Physiol Heart Circ Physiol, February 1, 2006; 290(2): H517 - H524. [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]
|
|
|
|
|
|
S. Janssens, P. Pokreisz, L. Schoonjans, M. Pellens, P. Vermeersch, M. Tjwa, P. Jans, M. Scherrer-Crosbie, M. H. Picard, Z. Szelid, et al. Cardiomyocyte-Specific Overexpression of Nitric Oxide Synthase 3 Improves Left Ventricular Performance and Reduces Compensatory Hypertrophy After Myocardial Infarction Circ. Res., May 14, 2004; 94(9): 1256 - 1262. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
E. N. Dedkova, X. Ji, Y. G. Wang, L. A. Blatter, and S. L. Lipsius Signaling Mechanisms That Mediate Nitric Oxide Production Induced by Acetylcholine Exposure and Withdrawal in Cat Atrial Myocytes Circ. Res., December 12, 2003; 93(12): 1233 - 1240. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.B. Massion, O. Feron, C. Dessy, and J.-L. Balligand Nitric Oxide and Cardiac Function: Ten Years After, and Continuing Circ. Res., September 5, 2003; 93(5): 388 - 398. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P B Massion and J-L Balligand Modulation of cardiac contraction, relaxation and rate by the endothelial nitric oxide synthase (eNOS): lessons from genetically modified mice J. Physiol., January 1, 2003; 546(1): 63 - 75. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Rubio and G. Ceballos Sole activation of three luminal adenosine receptor subtypes in different parts of coronary vasculature Am J Physiol Heart Circ Physiol, January 1, 2003; 284(1): H204 - H214. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. C L Bett, S. Dai, and D. L Campbell Cholinergic modulation of the basal L-type calcium current in ferret right ventricular myocytes J. Physiol., July 1, 2002; 542(1): 107 - 117. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Abi-Gerges, G. Szabo, A. S Otero, R. Fischmeister, and P.-F. Mery NO donors potentiate the {beta}-adrenergic stimulation of ICa,L and the muscarinic activation of IK,ACh in rat cardiac myocytes J. Physiol., April 15, 2002; 540(2): 411 - 424. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. H. M. Nascimento, L. Salle, J. Hoebeke, J. Argibay, and N. Peineau cGMP-mediated inhibition of cardiac L-type Ca2+ current by a monoclonal antibody against the M2 ACh receptor Am J Physiol Cell Physiol, October 1, 2001; 281(4): C1251 - C1258. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Chen, K. Spicher, M. Jiang, L. Birnbaumer, and G. T. Wetzel Lack of muscarinic regulation of Ca2+ channels in Gi2{alpha} gene knockout mouse hearts Am J Physiol Heart Circ Physiol, May 1, 2001; 280(5): H1989 - H1995. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Godecke, T. Heinicke, A. Kamkin, I. Kiseleva, R. H Strasser, U. K M Decking, T. Stumpe, G. Isenberg, and J. Schrader Inotropic response to {beta}-adrenergic receptor stimulation and anti-adrenergic effect of ACh in endothelial NO synthase-deficient mouse hearts J. Physiol., April 1, 2001; 532(1): 195 - 204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Nagata, C. Ye, M. Jain, D. S. Milstone, R. Liao, and R. M. Mortensen G{alpha}i2 but Not G{alpha}i3 Is Required for Muscarinic Inhibition of Contractility and Calcium Currents in Adult Cardiomyocytes Circ. Res., November 10, 2000; 87(10): 903 - 909. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Sasaki, K. Daitoku, A. Iwasa, and S. Motomura NO is involved in MCh-induced accentuated antagonism via type II PDE in the canine blood-perfused SA node Am J Physiol Heart Circ Physiol, November 1, 2000; 279(5): H2509 - H2518. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. Ullrich, M. Scherrer-Crosbie, K. D. Bloch, F. Ichinose, H. Nakajima, M. H. Picard, W. M. Zapol, and Z. M. N. Quezado Congenital Deficiency of Nitric Oxide Synthase 2 Protects Against Endotoxin-Induced Myocardial Dysfunction in Mice Circulation, September 19, 2000; 102(12): 1440 - 1446. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A.-M. Teichert, T. L. Miller, S. C. Tai, Y. Wang, X. Bei, G. B. Robb, M. J. Phillips, and P. A. Marsden In vivo expression profile of an endothelial nitric oxide synthase promoter-reporter transgene Am J Physiol Heart Circ Physiol, April 1, 2000; 278(4): H1352 - H1361. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. V.M. van Rijen, T. A.B. van Veen, M. M.P. Hermans, and H. J. Jongsma Human connexin40 gap junction channels are modulated by cAMP Cardiovasc Res, March 1, 2000; 45(4): 941 - 951. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 O.-E. Brodde and M. C. Michel Adrenergic and Muscarinic Receptors in the Human Heart Pharmacol. Rev., December 1, 1999; 51(4): 651 - 690. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. V. Brahmajothi and D. L. Campbell Heterogeneous Basal Expression of Nitric Oxide Synthase and Superoxide Dismutase Isoforms in Mammalian Heart : Implications for Mechanisms Governing Indirect and Direct Nitric Oxide-Related Effects Circ. Res., October 1, 1999; 85(7): 575 - 587. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J.-L. Balligand Regulation of cardiac {beta}-adrenergic response by nitric oxide Cardiovasc Res, August 15, 1999; 43(3): 607 - 620. [Full Text] [PDF]
|
|
|
|
 |
|
 C. E. Sears, J. K. Choate, and D. J. Paterson NO-cGMP pathway accentuates the decrease in heart rate caused by cardiac vagal nerve stimulation J Appl Physiol, February 1, 1999; 86(2): 510 - 516. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. J. JI, B. K. FLEISCHMANN, W. BLOCH, M. FEELISCH, C. ANDRESSEN, K. ADDICKS, and J. HESCHELER Regulation of the L-type Ca2+ channel during cardiomyogenesis: switch from NO to adenylyl cyclase-mediated inhibition FASEB J, February 1, 1999; 13(2): 313 - 324. [Abstract] [Full Text]
|
|
|
|
 |
|
 A. Kumar, R. Brar, P. Wang, L. Dee, G. Skorupa, F. Khadour, R. Schulz, and J. E. Parrillo Role of nitric oxide and cGMP in human septic serum-induced depression of cardiac myocyte contractility Am J Physiol Regulatory Integrative Comp Physiol, January 1, 1999; 276(1): R265 - R276. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X Han, L Kobzik, D Severson, and Y Shimoni Characteristics of nitric oxide-mediated cholinergic modulation of calcium current in rabbit sino-atrial node J. Physiol., June 15, 1998; 509(3): 741 - 754. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Han, I. Kubota, O. Feron, D. J. Opel, M. A. Arstall, Y.-Y. Zhao, P. Huang, M. C. Fishman, T. Michel, and R. A. Kelly Muscarinic cholinergic regulation of cardiac myocyte ICa-L is absent in mice with targeted disruption of endothelial nitric oxide synthase PNAS, May 26, 1998; 95(11): 6510 - 6515. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. E. Sears, J. K. Choate, and D. J. Paterson Inhibition of nitric oxide synthase slows heart rate recovery from cholinergic activation J Appl Physiol, May 1, 1998; 84(5): 1596 - 1603. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Vandecasteele, T. Eschenhagen, and R. Fischmeister Role of the NO-cGMP pathway in the muscarinic regulation of the L-type Ca2+ current in human atrial myocytes J. Physiol., February 1, 1998; 506(3): 653 - 663. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. G. Wang, C. E. Rechenmacher, and S. L. Lipsius Nitric Oxide Signaling Mediates Stimulation of L-Type Ca2+ Current Elicited by Withdrawal of Acetylcholine in Cat Atrial Myocytes J. Gen. Physiol., January 1, 1998; 111(1): 113 - 125. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Hu Nipavan Chiamvimonvat Toshio Yamagishi Eduardo Direct Inhibition of Expressed Cardiac L-Type Ca2+ Channels by S-Nitrosothiol Nitric Oxide Donors Circ. Res., November 19, 1997; 81(5): 742 - 752. [Abstract] [Full Text]
|
|
|
|
 |
|
 J.-L. Balligand and P. J. Cannon Nitric Oxide Synthases and Cardiac Muscle : Autocrine and Paracrine Influences Arterioscler Thromb Vasc Biol, October 1, 1997; 17(10): 1846 - 1858. [Abstract] [Full Text]
|
|
|
|
 |
|
 N. Abi-Gerges, T. Eschenhagen, L. Hove-Madsen, R. Fischmeister, and P.-F. Mery Methylene Blue Is a Muscarinic Antagonist in Cardiac Myocytes Mol. Pharmacol., September 1, 1997; 52(3): 482 - 490. [Abstract] [Full Text]
|
|
|
|
|
|
P. Musialek, M. Lei, H. F. Brown, D. J. Paterson, and B. Casadei Nitric Oxide Can Increase Heart Rate by Stimulating the Hyperpolarization-Activated Inward Current, If Circ. Res., July 19, 1997; 81(1): 60 - 68. [Abstract] [Full Text]
|
|
|
|
|
|
O. Feron, T. W. Smith, T. Michel, and R. A. Kelly Dynamic Targeting of the Agonist-stimulated m2 Muscarinic Acetylcholine Receptor to Caveolae in Cardiac Myocytes J. Biol. Chem., July 11, 1997; 272(28): 17744 - 17748. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Belhassen, O. Feron, D. M. Kaye, T. Michel, and R. A. Kelly Regulation by cAMP of Post-translational Processing and Subcellular Targeting of Endothelial Nitric-oxide Synthase (Type 3) in Cardiac Myocytes J. Biol. Chem., April 25, 1997; 272(17): 11198 - 11204. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Valenzuela, X. Han, U. Mende, C. Fankhauser, H. Mashimo, P. Huang, J. Pfeffer, E. J. Neer, and M. C. Fishman Galpha o is necessary for muscarinic regulation of Ca2+ channels in mouse heart PNAS, March 4, 1997; 94(5): 1727 - 1732. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. Kelly, J.-L. Balligand, and T. W. Smith Nitric Oxide and Cardiac Function Circ. Res., September 1, 1996; 79(3): 363 - 380. [Full Text]
|
|