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Molecular Pharmacology

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Abstract

Effect of cyclic GMP-increasing agents nitric oxide and C-type natriuretic peptide on bovine chromaffin cell function: inhibitory role mediated by cyclic GMP-dependent protein kinase.

F Rodriguez-Pascual, M T Miras-Portugal and M Torres
Molecular Pharmacology June 1996, 49 (6) 1058-1070;
F Rodriguez-Pascual
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M T Miras-Portugal
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M Torres
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Abstract

Both sodium nitroprusside (SNP), a nitric oxide (NO) generator, and C-type natriuretic peptide (CNP) have been found to raise cGMP levels in bovine chromaffin cells in a time- and concentration-dependent manner. The effect of these compounds on catecholamine secretion and calcium influx has also been studied, and both compounds were found to produce a slowly developing inhibitory effect on acetylcholine- or depolarization-stimulated catecholamine secretion and calcium increases without affecting the spontaneous release or the basal intracellular Ca2+ concentration. These inhibitory effects were observed only at high doses of acetylcholine or high levels of extracellular potassium and required concentrations of SNP or CNP very similar to those that increased cGMP levels. Preincubation with 100 microM zaprinast, a cGMP-phosphodiesterase inhibitor able to increase cGMP levels, mimicked the inhibitory effects of SNP and CNP. We investigated the effect of the soluble guanylate cyclase inhibitor methylene blue and the cGMP-dependent protein kinase (PKG) inhibitor 8-(4-chlorophenylthio)-guanosine 3',5'-cyclic monophosphorothioate, Rp isomer, on inhibition by SNP or CNP. Although methylene blue (10 microM) partially prevented the inhibitory effect of SNP, it did not do so for that produced by CNP, thus indicating that SNP acts through cGMP produced by the NO-activated guanylate cyclase. 8-(4-Chlorophenylthio)-guanosine 3',5'-cyclic monophosphorothioate, Rp isomer totally reversed both the SNP and CNP inhibitory effects. These results suggest that the activation of PKG mediates the inhibition induced by SNP and CNP. We successfully measured the PKG activity from cells preincubated with SNP or CNP, and our results show that this enzymatic activity increased with a time dependence very similar to the increase in the cGMP levels. Our results indicate that NO and CNP peptide inhibit secretagogue-stimulated catecholamine release via activation of soluble and particulate isoforms of the guanylate cyclase, respectively, presumably by inhibition of calcium entry through voltage-activated calcium channels. This inhibitory effects seems to be mediated by activation of the PKG.

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Molecular Pharmacology
Vol. 49, Issue 6
1 Jun 1996
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Abstract

Effect of cyclic GMP-increasing agents nitric oxide and C-type natriuretic peptide on bovine chromaffin cell function: inhibitory role mediated by cyclic GMP-dependent protein kinase.

F Rodriguez-Pascual, M T Miras-Portugal and M Torres
Molecular Pharmacology June 1, 1996, 49 (6) 1058-1070;

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Abstract

Effect of cyclic GMP-increasing agents nitric oxide and C-type natriuretic peptide on bovine chromaffin cell function: inhibitory role mediated by cyclic GMP-dependent protein kinase.

F Rodriguez-Pascual, M T Miras-Portugal and M Torres
Molecular Pharmacology June 1, 1996, 49 (6) 1058-1070;
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