Role of cyclic GMP‐dependent protein kinase in the contractile response to exogenous nitric oxide in rat cardiac myocytes
Open Access
- 1 April 2002
- journal article
- research article
- Published by Wiley in The Journal of Physiology
- Vol. 540 (2), 457-467
- https://doi.org/10.1113/jphysiol.2001.014126
Abstract
Nitric oxide (NO) can directly modulate cardiac contractility by accelerating relaxation and reducing diastolic tone. The intracellular mechanisms underlying these contractile effects are poorly understood. Here we investigate the role of cyclic GMP-dependent protein kinase (PKG) in the contractile response to exogenous NO in rat ventricular myocytes. Isolated ventricular myocytes were stimulated electrically and contractility was assessed by measuring cell shortening. Some cells were loaded with the fluorescent Ca2+ probe indo-1 AM for simultaneous assessment of the intracellular Ca2+ transient. The NO donor diethylamine NONOate (DEA/NO, 10 μm) significantly increased resting cell length, reduced twitch amplitude and accelerated time to 50 % relaxation (to 100.8 ± 0.2, 83.7 ± 3.0 and 88.9 ± 3.7 % of control values, respectively). The contractile effects of DEA/NO occurred without significant changes in the amplitude or kinetics of the intracellular Ca2+ transient, suggesting that the myofilament response to Ca2+ was reduced. These effects were abolished by inhibition of either guanylyl cyclase (with 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one; ODQ, 10 μm) or PKG (with Rp-8-Br-cGMPs, 10 μm) suggesting that, at the concentration investigated, the effects of DEA/NO were mediated exclusively by PKG, following activation of guanylyl cyclase and elevation of cGMP. Direct activation of PKG with 8-pCPT-cGMP (10 μm) mimicked the effects of DEA/NO (resting cell length and time to 50 % relaxation were 100.6 ± 0.1 and 90.5 ± 1.5 % of control values, respectively).The reduced myofilament Ca2+ responsiveness was not attributable to an intracellular acidosis since the small reduction in pHi induced by DEA/NO was found to be uncoupled from its contractile effects. However, hearts treated with DEA/NO (10 μm) showed a significant increase (1.4-fold; P < 0.01) in troponin I phosphorylation compared to control, untreated hearts. These results suggest that the reduction in myofilament Ca2+ responsiveness produced by DEA/NO results from phosphorylation of troponin I by PKG.This publication has 50 references indexed in Scilit:
- Cardiac contractile impairment associated with increased phosphorylation of troponin I in endotoxemic ratsThe FASEB Journal, 2000
- Subcellular mechanisms of the positive inotropic effect of angiotensin II in cat myocardiumThe Journal of Physiology, 2000
- Inhibition of nitric oxide synthase augments the positive inotropic effect of nitric oxide donors in the rat heartThe Journal of Physiology, 2000
- Peroxynitrite is a positive inotropic agent in atrial and ventricular fibres of the frog heartThe Journal of Physiology, 1999
- Impaired cardiomyocyte relaxation and diastolic function in transgenic mice expressing slow skeletal troponin I in the heartThe Journal of Physiology, 1999
- Activation of Nitric Oxide Synthase (NOS3) by Mechanical Activity Alters Contractile Activity in a Ca2+-Independent Manner in Cardiac Myocytes: Role of Troponin I PhosphorylationBiochemical and Biophysical Research Communications, 1999
- Paracrine modulation of heart cell function by endothelial cellsCardiovascular Research, 1996
- Cyclic GMP-Mediated Phospholamban Phosphorylation in Intact CardiomyocytesBiochemical and Biophysical Research Communications, 1995
- cGMP‐dependent protein kinase decreases calcium sensitivity of skinned cardiac fibersFEBS Letters, 1982
- Calcium sensitivity of the contractile system and phosphorylation of troponin in hyperpermeable cardiac cells.The Journal of general physiology, 1980