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Alpha1-adrenoceptor-mediated inhibition of cellular cAMP accumulation in neonatal rat ventricular myocytes

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Summary

We studied adrenergic regulation of cellular cAMP in neonatal rat ventricular myocytes. Since CAMP content depends on synthesis, breakdown and egress, the contribution of each of these mechanisms was assessed. In the presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine, cAMP accumulation stimulated by the β-adrenoceptor agonist (−)-isoprenaline was diminished when the mixed α + β adrenoceptor agonist (−)-noradrenaline was coincubated with (−)-isoprenaline. Moreover, adenylyl cyclase activation stimulated by (−)-isoprenaline was decreased by (−)-noradrenaline and by the selective a1-adrenoceptor agonists (−)-phenylephrine and methoxamine, suggesting that α-adrenoceptor agonism regulates CAMP metabolism through its effect on the synthetic pathway. Evidence for α1-adrenoceptor mediation of this response was enhancement of (−)-noradrenaline-induced cAMP generation by the selective α1-adrenoceptor antagonist terazosin (10 nmol/l). The selective α2-adrenoceptor antagonist yohimbine (10 nmol/l) had no effect. The α1-adrenoceptor mediated depression of (−)-isoprenaline-stimulated CAMP generation and adenylyl cyclase activation was prevented by terazosin and in separate experiments markedly enhanced by pertussis toxin pretreatment, suggesting involvement of a guanine-nucleotide regulatory protein in this process.

Occupation of the α1-adrenoceptor by (−)-noradrenaline did not accelerate the rate of CAMP breakdown in the absence of phosphodiesterase inhibition. Furthermore, there was no enhancement of total phosphodiesterase activity by (−)-noradrenaline in the presence of (−)-propranolol. By contrast, pertussis toxin pretreatment augmented phosphodiesterase activity. Neither pertussis toxin nor (−)-noradrenaline increased CAMP egress.

We conclude that in rat neonatal cardiac myocytes agonist occupation of the α1-adrenoceptor inhibits β-adrenoceptor stimulated CAMP accumulation most likely by coupling to a guanine nucleotide inhibitory protein.

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References

  • Alvarez R, Daniels DV (1990) A single column method for the assay of adenylate cyclase. Anal Biochem 187:98–103

    Google Scholar 

  • Barber R, Butcher RW (1983) The egress of cyclic AMP from metazoan cells. Adv Cyclic Nuc Res 15:119–138

    Google Scholar 

  • Boltwood CM Jr, Appleyard RF, Glantz SA (1989) Left ventricular volume measurement by conductance catheter in intact dogs. Circulation 80:1360–1377

    Google Scholar 

  • Buxton ILO, Brunton LL (1985) Action of the cardiac α1-adrenergic receptor. J Biol Chem 26:6733–6737

    Google Scholar 

  • Buxton ILO, Brunton LL, (1986) α-Adrenergic receptors on rat ventricular myocytes: characteristics and linkage to CAMP metabolism. Am J Physiol 251:H307-H313

    Google Scholar 

  • Bray P, Carter A, Guo V, Puckett C, Kamholz J, Spiegel A, Nirenberg M (1987) Human cDNA clones for an α subunit of Gi signal-transduction protein. Proc Natl Acad Sci USA 84:5115–5119

    Google Scholar 

  • Brunton LL, Heasley LE (1988) cAMP export and its regulation by prostaglandin A1. Methods Enzymol 159:83–90

    Google Scholar 

  • Culling W, Penny WJ, Cunliffe G, Flores NA, Sheridan DJ (1987) Arrhythmogenic and electrophysiological effects of alpha adrenoceptor stimulation during myocardial ischemia and reperfusion. J Mol Cell Cardiol 19:251–258

    Google Scholar 

  • Dietz R, Schömig A, Strasser R, Mauerer W Manthey J, Kübler W (1981) Humoral adaptation mechanism in heart failure. In: Goodwin JF, Hjalmarson A, Olsen EGJ (eds) Congestive cardiomyopathy. AB Hassle, Molndal, pp 128–137

    Google Scholar 

  • Endoh M, Hiramoto T, Ishihata A, Takanashi M, Inui J (1991) Myocardial α1-adrenoceptors mediate positive inotropic effect and changes in phosphatidylinositol metabolism. Circ Res 68:1179–1190

    Google Scholar 

  • Endoh M, Takanashi M, Norota I (1992) Role of α1-adrenoceptor subtype in production of the positive inotropic effect mediated via myocardial α1-adrenoceptors in the rabbit papillary muscle: influence of selective α1-subtype antagonists WB4101 and 5-methylurapidil. Naunyn-Schmiedeberg's Arch Pharmacol 348:578–585

    Google Scholar 

  • Filburn CR, Karn J (1973) An isotopic assay of cyclic 3′,5′-nucleotide phosphodiesterase with aluminum oxide columns. Anal Biochem 52:505–516

    Google Scholar 

  • Fisher RA, Harary I, Thomas JA (1984) Homologous desensitization of rat heart cells to β1-adrenergic stimulation and the absence of α1-adrenergic or prostaglandin E1 effects. J Cyclic Nucl Protein Phos Res 9:449–460

    Google Scholar 

  • Fleming JW, Watanabe AM (1988) Muscarinic cholinergic-receptor in stimulation of specific GTP hydrolysis related to adenylate cyclase activity. Circ Res 64:340–350

    Google Scholar 

  • Fleming JW, Strawbridge RA, Watanabe AM (1987) Muscarinic receptor regulation of cardiac adenylate cyclase activity. J Mol Cell Cardiol 19:47–61

    Google Scholar 

  • Fleming JW, Hodges TD, Watanabe AM (1988) Pertussis toxin-treated dog: a whole animal model of impaired inhibitory regulation of adenylate cyclase. Circ Res 62:992–1000

    Google Scholar 

  • Folkow B, Haggendal J, Lisander B (1967) Extent of release and elimination of noradrenaline at peripheral adrenergic nerve terminals. Acta Physiol Scand [Suppl] 307:1–38

    Google Scholar 

  • Foster KA, McDermott PJ, Robishaw JD (1990) Expression of G proteins in rat cardiac myocytes: effect of KCl depolarization. Am J Physiol 259:H432-H441

    Google Scholar 

  • Freissmuth M, Casey PJ, Gilman AG (1989) G proteins control diverse pathways of transmembrane signaling. FASEB J 3:2125–2131

    Google Scholar 

  • Gambassi G, Berenholtz S, Ziman B, Lakatta EG, Capogrossi C (1991) Opposing effects of α1A and α1B receptors on the inotropic response to α1-adrenergic stimulation in adult rat myocytes. Circulation [Supp II]84:403

    Google Scholar 

  • Gettys TW Blackmore PF, Redmon JB, Beebe SJ, Corbin JD (1987) Short-term feedback regulation of CAMP by accelerated degradation in rat tissues. J Biol Chem 262:333–339

    Google Scholar 

  • Gettys TW Vine AJ, Simonds MF, Corbin JD (1988) Activation of the particulate low Km-phosphodiesterase of adipocytes by addition of cAMP-dependent protein kinase. J Biol Chem 263:10359–10363

    Google Scholar 

  • Glantz SA, Slinker BK (1990) Primer of applied regression and analysis of variance. New York, McGraw Hill, Chapt 9 and 10

    Google Scholar 

  • Han H-M, Robinson RB, Bilezikian JP, Steinberg SF (1989) Developmental changes in guanine nucleotide regulatory proteins in the rat myocardial α1-adrenergic receptor complex. Circ Res 65:1763–1773

    Google Scholar 

  • Hazeki O, Ui M (1981) Modification by islet-activating protein of receptor-mediated regulation of cyclic AMP accumulation in isolated rat heart cells. J Biol Chem 2856–2862

  • Heinrich CL, Simpson PC (1988) Differential acute and chronic responses of protein kinase C in cultured neonatal rat heart myocytes to α1-adrenergic and phorbol ester stimulation. J Mol Cell cardiol 20:1081–1085

    Google Scholar 

  • Hescheler J, Rosenthal W, Trautwein W, Schultz G (1987) The GTP-binding protein, Go, regulates neuronal calcium channels. Nature 325:445–447

    Google Scholar 

  • Hilal-Dandan R, Urasawa K, Brunton LL (1992) Endothelin inhibits adenylate cyclase and stimulates phosphoinositide hydrolysis in adult cardiac myocytes. J Biol Chem 267:10620–10624

    Google Scholar 

  • Im M-J, Graham RM (1990a) A novel guanine nucleotide-binding protein coupled to the α1-adrenergic receptor. I. Identification by photoblabeling of membrane and ternary complex preparations. J Biol Chem 265:18944–18951

    Google Scholar 

  • Im M-J, Riek RP, Graham RM (1990b) A novel guanine nucleotide-binding protein coupled to the α1-adrenergic receptor. II. Purification, characterization, and reconstitution. J Biol Chem 265:18944–18951

    Google Scholar 

  • Jones DT, Reed RR (1987) Molecular cloning of five GTP-binding protein cDNA species from rat olfactory neuroepithelium. J Biol Chem 262:14241–14249

    Google Scholar 

  • Karliner JS, Barnes P, Hamilton CA, Dollery CT (1979) Alpha-adrenergic receptors in guinea pig nyocardium: Identification by binding of a new radioligand (3H)-prazosin. Biochem Biophys Res Commun 90:142–148

    Google Scholar 

  • Karliner JS, Motulsky HJ, Dunlap J, Brown JH, Insel PA (1982) Verapamil competitively inhibits alpha-adrenergic and muscarinic but not beta-adrenergic receptors in rat myocardium. J Cardiovas Pharmacol 4:515–520

    Google Scholar 

  • Karliner JS, Simpson P, Taylor JE (1985) Adrenergic receptor characteristics of cardiac myocytes cultured in serum-free medium in comparison with serum supplemented medium. Biochem Biophys Res Comm 128:376–382

    Google Scholar 

  • Karliner JS, Simpson P, Honbo N, Woloszyn W (1986) Mechanisms and time course of β1-receptor desensitization in mammalian cardiac myocytes. Cardiovasc Res 20:221–228

    Google Scholar 

  • Karliner JS, Kagiya T, Simpson PC (1990) Effects of pertussis toxin on α1-agonist-mediated phosphotidylinositide turnover and myocardial cell hypertrophy in neonatal rat ventricular myocytes. Exerientia 46:81–84

    Google Scholar 

  • Katada T, Gilman AG, Watanabe Y, Bauer S, Jakobs KH (1985) Protein kinase C phosphorylates the inhibitory guanine nucleotide-binding regulatory component and apparently suppresses its function in hormonal inhibition of adenylyl cyclase. Eur J Biochem 85:431–437

    Google Scholar 

  • Keely SL, Corbin JD, Lincoln T (1977) Alpha adrenergic involvement in heart metabolism: effects on adenosine cyclic 3′,5′-monophosphate-dependent protein kinase, guanosine cyclic 3′,5′-monophosphate, and glucose transport. Mol Pharmacol 13:965–975

    Google Scholar 

  • Keung E, Karliner JS (1990) Complex regulation of calcium current in cardiac cells: dependence on a pertussis toxin sensitive substrate, ATP and an α1-adrenoceptor. J Clin Invest 95:950–954

    Google Scholar 

  • Levitzki A (1987) Regulation of hormone-sensitive adenylate cyclase. Trends Pharmacol Sci 8:299–303

    Google Scholar 

  • Liang BT, Galper JB (1988) Differential sensitivity of αo and αi to ADP-ribosylation by pertussis toxin in the intact cultured embryonic chick ventricular myocyte. Biochem Pharmacol 37:4549–4555

    Google Scholar 

  • Limbird LE (1988) Receptors linked to inhibition of adenylate cyclase: additional signaling mechanisms. FASEB J 2:2686–2695

    Google Scholar 

  • Lowry OH, Rosenbrough NJ, Farn AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275

    Google Scholar 

  • Madelian V, Shain W (1987) Regulation of isoproterenol-induced cyclic AMP accumulation in LRM55 glial cells by phosphodiesterase. J Pharmacol Exp Ther 243:618–623

    Google Scholar 

  • McPherson GA (1985) Analysis of radioligand binding experiments. A collection of computer programs for the IBM PC. J Pharmacol Methods 14:213–228

    Google Scholar 

  • Meeker RB, Harden TK (1982) Muscarinic cholinergic receptor-mediated activation of phosphodiesterase. Mol Pharmacol 22:310–319

    Google Scholar 

  • Miot R, Erneux C, Wells JN, Dumont JE (1984) The effects of alkylated xanthines on cyclic AMP accumulation in dog thyroid slices exposed to carbamylcholine. Mol Pharmacol 25:261–266

    Google Scholar 

  • Moss J (1987) Signal transduction by receptor-responsive guanyl nucleotide-binding proteins: modulation by bacterial toxin-catalyzed ADP-ribosylation. Clin Res 35:451–458

    Google Scholar 

  • Munson PF; Rodbard D (1980) LIGAND: a versatile computerized approach for characterization of ligand-binding systems. Anal Biochem 107:220–239

    Google Scholar 

  • Neer EJ, Clapham DE (1988) Roles of G protein subunits in transmembrane signalling. Nature 333:129–134

    Google Scholar 

  • Rocha-Singh KJ, Hines DW, Honbo NY, Karliner JS (1991) Concanavalin A amplifies both β-adrenergic and muscarinic cholinergic receptor-adenylate cyclase-linked pathways in cardiac myocytes. J Clin Invest 88:760–766

    Google Scholar 

  • Scatchard G (1949) The attraction of proteins for small molecules and ions. Ann New York Acad Sci 51:660–672

    Google Scholar 

  • Scholz J, Schaefer B, Schmitz W, Scholz H, Steinfath M, Lohse M, Schwabe U, Puurunen J (1988) Alpha1-adrenoceptor-mediated positive inotropic effect and inositol phosphate increase in mammalian heart. J Pharmacol Exp Ther 245:327–335

    Google Scholar 

  • Simon MI, Strathmann MP, Gautam N (1991) Diversity of G proteins in signal transduction. Science 252:802–808

    Google Scholar 

  • Simonds WF, Goldsmith PK, Codina J, Unson CG, Spiegel AM (1989) Gi2 mediates α2-adrenergic inhibition of adenylyl cyclase in platelet membranes: In situ identification with Gα C-terminal antibodies. Proc Natl Acad Sci USA 86:7809–7813

    Google Scholar 

  • Simpson P (1983) Norepinephrine-stimulated hypertrophy of cultured rat myocardial cells is an α1-adrenergic response. J Clin Invest 72:732–738

    Google Scholar 

  • Simpson PC, Kariya K-I, Karns LR, Long CS, Karliner JS (1991) Adrenergic hormones and control of cardiac myocyte growth. Mol Cell Biochem 104:35–43

    Google Scholar 

  • Starke K (1981) α-Adrenoceptor subclassification. Rev Physiol Biochem Pharmacol 88:199–236

    Google Scholar 

  • Steinberg SF, Kaplan LM, Inouye T, Zhang JF, Robinson RB (1989) Alpha1-adrenergic stimulation of 1,4,5-inositol trisphosphate formation in ventricular myocytes. J Pharmacol Exp Ther 250:1141–1148

    Google Scholar 

  • Steiner AL, Parker CW, Kipnis DM (1972) Radioimmunoassay for cyclic nucleotides. J Biol Chem 247:1106–1113

    Google Scholar 

  • Swinnen JV, Joseph DR, Conti M (1989) The mRNA encoding a high-affinity cAMP phosphodiesterase is regulated by hormones and cAMP. Proc Natl Acad Sci USA 86:8197–8201

    Google Scholar 

  • Takanashi M, Norota I, Endoh M (1992) Potential inhibitory action of chlorethylclonidine on the positive inotropic effect and phosphoinositide hydrolysis mediated via myocardial α1-adrenoceptors in the rabbit ventricular myocardium. Naunyn-Schmiedeberg's Arch Pharmacol 343:669–673

    Google Scholar 

  • Tanner LI, Harden TK, Wells JN, Martin MW (1986) Identification of the phosphodiesterase regulated by muscarinic cholinergic receptors of 1321N1 human astrocytoma cells. Mol Pharmacol 29:455–460

    Google Scholar 

  • Triggle CR, Triggle DJ (1976) Structure and function of the synapse. In: Triggle DJ, Triggle CR (eds) Chemical pharmacology of the synapse. Academic Press, London, pp 1–127

    Google Scholar 

  • Wang X-L, Wettwer E, Gross G, Ravens U (1991) Reduction of cardiac outward currents by alpha-1 adrenoceptor stimulation: A subtype-specific effect? J Pharmacol Exp Ther 259:783–788

    Google Scholar 

  • Watanabe AM, Hathaway DR, Besch HR Jr, Farmer BB, Harris RA (1977) α-Adrenergic reduction of cyclic adenosine monophosphate concentrations in rat myocardium. Circ Res 40:596–602

    Google Scholar 

  • Weber HW, Chung F-Z, Day K, Appleman MM (1987) Insulin stimulation of cyclic AMP phosphodiesterase is independent from the G-protein pathways involved in adenylate cyclase regulation. J Cyclic Nucl Protein Phos Res 11:345–354

    Google Scholar 

  • Weiss S, Kemp DE, Lenox RH, Ellis J (1987) α2-Adrenergic receptors mediate inhibition of cyclic AMP production in neurons in primary culture. Brain Res 414:390–394

    Google Scholar 

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Authors and Affiliations

  1. Cardiology Section of the Veterans Affairs Medical Center, San Francisco

    Sondra Barrett, Norman Honbo & Joel S. Karliner

  2. Cardiovascular Research Institute and the Department of Medicine, University of California, San Francisco, USA

    Sondra Barrett, Norman Honbo & Joel S. Karliner

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  1. Sondra Barrett
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  2. Norman Honbo
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Supported by a grant from the Department of the Veterans Affairs Research Service and Program Project Grant HL 25847 from the National Heart, Lung and Blood Institute

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Barrett, S., Honbo, N. & Karliner, J.S. Alpha1-adrenoceptor-mediated inhibition of cellular cAMP accumulation in neonatal rat ventricular myocytes. Naunyn-Schmiedeberg's Arch Pharmacol 347, 384–393 (1993). https://doi.org/10.1007/BF00165388

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