Particulate guanylyl cyclase (pGC) and soluble guanylyl cyclase (sGC) are cGMP-generation systems distributed in different intracellular locations. Our aim was to test the hypothesis that the functional effects of cGMP produced by pGC and sGC on contraction and Ca2+ transients would differ in ventricular myocytes. We measured myocyte shortening from adult mice using a video edge-detector and investigated the functional changes after stimulating pGC with C-type natriuretic peptide (CNP; 10–8 M and 10–7 M) or sGC with S-nitroso-N-acetyl-penicillamine (SNAP; nitric oxide donor; 10–6 M and 10–5 M). Significant concentration-dependent decreases in percentage shortening (PCS), maximal rate of shortening (RSmax), and relaxation (RRmax) were produced by CNP. To a similar degree, SNAP concentration-dependently reduced PCS, RSmax, and RRmax. The addition of Rp-8-[(4-chlorophenyl)thio]-cGMPS triethylamine (cGMP-dependent protein kinase inhibitor; 5 × 10–6 M) or erythro-9-(2-hydroxy-3-nonyl) adenine (cGMP-stimulated cAMP phosphodiesterase inhibitor; 10–5 M) reduced the responses induced by CNP or SNAP, suggesting that their actions were through cGMP-mediated pathways. While SNAP significantly increased intracellular cGMP concentration by 57%, CNP had little effect on cGMP production. We also found that CNP markedly decreased the amplitude of Ca2+ transients while SNAP had little effect, suggesting the cGMP generated by sGC may decrease myofilament Ca2+ sensitivity. The small amount of cGMP generated by pGC had a major effect in reducing Ca2+ level. This study suggested the existence of compartmentalization for cGMP in ventricular myocytes.
ShahAM, SpurgeonHA, SollottSJ, TaloA, LakattaEG. 8-bromo-cGMP reduces the myofilament response to Ca2+ in intact cardiac myocytes. Circ Res74: 970–978, 1994.
2.
MuradF. The nitric oxide-cyclic GMP signal transduction system for intracellular and intercellular communication. Recent Prog Horm Res49: 239–248, 1994.
3.
ShahAM, MacCarthyPA. Paracrine and autocrine effects of nitric oxide on myocardial function. Pharmacol Ther86: 49–86, 2000.
4.
LaylandJ, LiJM, ShahAM. Role of cyclic GMP-dependent protein kinase in the contractile response to exogenous nitric oxide in rat cardiac myocytes. J Physiol540: 457–467, 2002.
5.
KojdaG, KottenbergK. Regulation of basal myocardial function by NO. Cardiovasc Res41: 514–523, 1999.
6.
LucasKA, PitariGM, KazerounianS, Ruiz-StewartI, ParkJ, SchulzS, ChepenikKP, WaldmanSA. Guanylyl cyclases and signaling by cyclic GMP. Pharmacol Rev52: 375–414, 2000.
7.
FrancisSH, TurkoIV, CorbinJD. Cyclic nucleotide phosphodiesterases: relating structure and function. Prog Nucleic Acid Res Mol Biol65: 1–52, 2001.
8.
RhoEH, PerkinsWJ, LorenzRR, WarnerDO, JonesKA. Differential effects of soluble and particulate guanylyl cyclase on Ca2+ sensitivity in airway smooth muscle. J Appl Physiol92: 257–263, 2002.
9.
HartCY, HahnEL, MeyerDM, BurnettJCJr., RedfieldMM. Differential effects of natriuretic peptides and NO on LV function in heart failure and normal dogs. Am J Physiol281:H146–H154, 2001.
10.
ZolleO, LawrieAM, SimpsonAW. Activation of the particulate and not the soluble guanylate cyclase leads to the inhibition of Ca2+ extrusion through localized elevation of cGMP. J Biol Chem275: 25892–25899, 2000.
11.
SuJ, ZhangS, TseJ, ScholzPM, WeissHR. Alterations in nitric oxide/cyclic GMP pathway in ventricular myocytes from obese leptin-deficient mice. Am J Physiol285:H2111–H2117, 2003.
12.
ZhangQ, MolinoB, YanL, HaimT, VaksY, ScholzPM, WeissHR. Nitric oxide and cGMP protein kinase activity in aged ventricular myocytes. Am J Physiol281:H2304–H2309, 2001.
13.
GongGX, WeissHR, TseJ, ScholzPM. Exogenous nitric oxide reduces oxygen consumption of isolated ventricular myocytes less than other forms of guanylate cyclase stimulation. Eur J Pharmacol344: 299–305, 1998.
14.
PierkesM, GambaryanS, BoknikP, LohmannSM, SchmitzW, PotthastR, HoltwickR, KuhnM. Increased effects of C-type natriuretic peptide on cardiac ventricular contractility and relaxation in guanylyl cyclase A-deficient mice. Cardiovasc Res53: 852–861, 2002.
15.
NirA, ZhangDF, FixlerR, BurnettJCJr., EilamY, HasinY. C-type natriuretic peptide has a negative inotropic effect on cardiac myocytes. Eur J Pharmacol412: 195–201, 2001.
16.
HiroseM, FurukawaY, KurogouchiF, NakajimaK, MiyashitaY, ChibaS. C-type natriuretic peptide increases myocardial contractility and sinus rate mediated by guanylyl cyclase-linked natriuretic peptide receptors in isolated, blood-perfused dog heart preparations. J Pharmacol Exp Ther286: 70–76, 1998.
17.
ChampionHC, SkafMW, HareJM. Role of nitric oxide in the pathophysiology of heart failure. Heart Fail Rev8: 35–46, 2003.
18.
ZioloMT, KatohH, BersDM. Positive and negative effects of nitric oxide on Ca2+ sparks: influence of beta-adrenergic stimulation. Am J Physiol281:H2295–H2303, 2001.
19.
StoneJR, MariettaMA. Soluble guanylate cyclase from bovine lung: activation with nitric oxide and carbon monoxide and spectral characterization of the ferrous and ferric states. Biochemistry33: 5636–5640, 1994.
20.
van CalkerD, MullerM, HamprechtB. Adenosine inhibits the accumulation of cyclic AMP in cultured brain cells. Nature276: 839–841, 1978.
21.
GauthierNS, HeadrickJP, MatherneGP. Myocardial function in the working mouse heart overexpressing cardiac A1 adenosine receptors. J Mol Cell Cardiol30: 187–193, 1998.
22.
StepinskiJ, WendtU, LewkoB, AngielskiS. Co-operation between particulate and soluble guanylyl cyclase systems in the rat renal glomeruli. J Physiol Pharmacol51: 497–511, 2000.
23.
HamadAM, RangeS, HollandE, KnoxAJ. Regulation of cGMP by soluble and particulate guanylyl cyclases in cultured human airway smooth muscle. Am J Physiol273:L807–L813, 1997.
24.
NeysesL, VetterH. Action of atrial natriuretic peptide and angiotensin II on the myocardium: studies in isolated rat ventricular cardiomyocytes. Biochem Biophys Res Commun163: 1435–1443, 1989.
25.
SumiiK, SperelakisN. cGMP-dependent protein kinase regulation of the L-type Ca2+ current in rat ventricular myocytes. Circ Res77: 803–812, 1995.
26.
JiangLH, GawlerDJ, HodsonN, MilliganCJ, PearsonHA, PorterV, WrayD. Regulation of cloned cardiac L-type calcium channels by cGMP-dependent protein kinase. J Biol Chem275: 6135–6143, 2000.
27.
ShenJB, PappanoAJ. On the role of phosphatase in regulation of cardiac L-type calcium current by cyclic GMP. J Pharmacol Exp Ther301: 501–506, 2002.
28.
HerzigS, NeumannJ. Effects of serine/threonine protein phosphatases on ion channels in excitable membranes. Physiol Rev80: 173–210, 2000.
29.
RaeymaekersL, HofmannF, CasteelsR. Cyclic GMP-dependent protein kinase phosphorylates phospholamban in isolated sarcoplasmic reticulum from cardiac and smooth muscle. Biochem J252: 269–273, 1998.
30.
ZhangR, ZhaoJ, PotterJD. Phosphorylation of both serine residues in cardiac troponin I is required to decrease the Ca2+ affinity of cardiac troponin C. J Biol Chem270: 30773–30780, 1995.
31.
YamakageM, HirshmanCA, CroxtonTL. Sodium nitroprusside stimulates Ca2+-activated K+ channels in porcine tracheal smooth muscle cells. Am J Physiol270:L338–L345, 1996.
32.
LanglandsJM, DiamondJ. The effect of phenylephrine on inositol 1,4,5-trisphosphate levels in vascular smooth muscle measured using a protein binding assay system. Biochem Biophys Res Commun173: 1258–1265, 1990.
33.
BrusqJM, MayouxE, GuiguiL, KirilovskyJ. Effects of C-type natriuretic peptide on rat cardiac contractility. Br J Pharmacol128: 206–212, 1999.
34.
MurthyKS, MakhloufGM. Identification of the G protein-activating domain of the natriuretic peptide clearance receptor (NPR-C). J Biol Chem274: 17587–17592, 1999.
35.
Vila-PetroffMG, YounesA, EganJ, LakattaEG, SollottSJ. Activation of distinct cAMP-dependent and cGMP-dependent pathways by nitric oxide in cardiac myocytes. Circ Res84: 1020–1031, 1999.
36.
KayeDM, WiviottSD, KellyRA. Activation of nitric oxide synthase (NOS3) by mechanical activity alters contractile activity in a Ca2+-independent manner in cardiac myocytes: role of troponin I phosphorylation. Biochem Biophys Res Commun256: 398–403, 1999.
