Ca(++)-dependent constitutive nitric oxide synthase is not involved in the cyclic GMP-increasing effects of carbachol in ventricular cardiomyocytes.
We investigated the effects of the M-cholinoceptor agonist carbachol on cyclic GMP (cGMP) content and contractile response in the absence and presence of the nitric oxide synthase inhibitor NG-nitro-L-arginine in guinea-pig isolated ventricular cardiomyocytes. Carbachol (10 mumol/l, 10 min) increased basal cGMP content to approximately 200% and contractile response to 118%. Preincubation of the cardiomyocytes with NG-nitro-L-arginine (0.1 mumol/l, 60 min) did not alter the effects of carbachol on neither cGMP content or contractile response. Moreover, nitric oxide synthase activity was undetectable in crude or ADP-agarose purified cytosolic and particulate fractions of homogenized isolated ventricular cardiomyocytes. Pretreatment with pertussis toxin did not affect the carbachol-mediated increase in cGMP content or contractile response. However, methylene blue abolished the elevation in cGMP content by carbachol, without changing contractile response. It is concluded that the carbachol-mediated increase in cGMP content and contractile response in ventricular cardiomyocytes is neither mediated via a nitric oxidebiosynthesis pathway nor via a pertussis toxin-sensitive GTP-binding protein. Furthermore, the cGMP increase by carbachol is due to an activation of soluble guanylyl cyclase and is dissociated from the contractile response. We therefore assume that carbachol activates two independent effector cascades, one leading to an elevation in cGMP content and the other to an increase in contractile response and that none of the effects are mediated via endogenous nitric oxide formation.
Stein
B
B
Abteilung Allgmeine Pharmakologie, Universitäts-Krankenhaus Eppendorf, Hamburg, F.R.G.
Drögemüller
A
A
Mülsch
A
A
Schmitz
W
W
Scholz
H
H
D000594:Amino Acid Oxidoreductases / Q000502:physiology*; D000818:Animals; D002118:Calcium / Q000502:physiology*; D002217:Carbachol / Q000494:pharmacology*; D006152:Cyclic GMP / Q000096:biosynthesis*; D019204:GTP-Binding Proteins / Q000502:physiology; D006168:Guinea Pigs; D066298:In Vitro Techniques; D008751:Methylene Blue / Q000494:pharmacology; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000378:metabolism*; D019001:Nitric Oxide Synthase; D037342:Pertussis Toxin; D011621:Pteridines / Q000494:pharmacology; D011622:Pterins; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D011621:Pteridines; D011622:Pterins; D010566:Virulence Factors, Bordetella; D002217:Carbachol; C016727:sepiapterin; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D006152:Cyclic GMP; D002118:Calcium; D008751:Methylene Blue