Abstract
We injected rats with pertussis toxin, known to cause ADP ribosylation of the Gi regulatory protein of the adenylate cyclase complex and of another closely related GTP binding protein in the heart, and after 7 days we examined several effects of muscarinic activation on the heart. The negative chronotropic effect of carbamoylcholine on spontaneously beating perfused hearts was conspicuously diminished. While 10−5 mol/l carbamoylcholine invariably produced heart arrest in control rats, the heart rate did not decrease by more than 20% in the toxin-treated rats even when the concentration of carbamoylcholine was raised to 10−2 mol/l. The negative inotropic effect of carbamoylcholine examined on electrically paced ventricles perfused with isoproterenol was reduced, while the maximum positive inotropic effect of isoproterenol was substantially increased after toxin treatment. The inhibitory action of carbamoylcholine on the isoproterenol-stimulated accumulation of cyclic AMP in the heart auricles was attenuated. The weakening by pertussis toxin of the negative inotropic effect of carbamoylcholine is probably mainly due to the ADP ribosylation of the Gi regulatory protein and the subsequent loss of influence of muscarinic receptors on adenylate cyclase. The blockade of the negative chronotropic action of carbamoylcholine by pertussis toxin strongly indicates, together with other recently published evidence, that the Gi or another closely related GTP binding protein in the cardiac pacemaker cells is involved in the coupling of muscarinic receptors to the K+ channels.
Similar content being viewed by others
References
Biegon RL, Epstein PM, Pappano AJ (1980) Muscarinic antagonism of the effects of a phosphodiesterase inhibitor (methylisobutylxanthine) in embryonic chick ventricles. J Pharmacol Exp Ther 215:348–356
Birdsall NJM, Berrie CP, Burgen ASV, Hulme EC (1980) Modulation of the binding properties of muscarinic receptors: Evidence for receptor-effector coupling. In: Pepeu G, Kuhar M, Enna SJ (eds) Receptors for neurotransmitters and peptide hormones. Raven Press, New York, pp 107–116
Breitwieser GE, Szabo G (1985) Uncoupling of cardiac muscarinic and beta-adrenergic receptors from ion channels by a guanine nucleotide analogue. Nature 317:538–540
Brown JH (1979) Cholinergic inhibition of catecholamine-stimulable cyclic AMP accumulation in murine atria. J Cyclic Nucleotide Protein Phosphor Res 5:423–433
Brown HF (1982) Electrophysiology of the sinoatrial node. Physiol Rev 62:505–530
Creazzo T, Titus L, Hartzell C (1983) Neural regulation of the heart. A model for modulation of voltage-sensitive channels and regulation of cellular metabolism by neurotransmitters. Trends Neurol Sci 6:430–433
Endoh M, Maruyama M, Iijima T (1985) Attenuation of muscarinic cholinergic inhibition by islet-activating protein in the heart. Am J Physiol 249:H309-H320
Florio VA, Sternweis PC (1985) Reconstitution of resolved muscarinic cholinergic receptors with purified GTP-binding proteins. J Biol Chem 260:3477–3483
Gilman AG (1984) G proteins and dual control of adenylate cyclase. Cell 36:577–579
Haga K, Haga T, Ichiyama A, Katada T, Kurose H, Ui M (1985) Functional reconstitution of purified muscarinic receptors and inhibitory guanine nucleotide regulatory protein. Nature 316:731–733
Halvorsen SW, Nathanson NM (1984) Ontogenesis of physiological responsiveness and guanine nucleotide sensitivity of cardiac muscarinic receptors during chick embryonic development. Biochemistry 23:5813–5821
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 256:2856–2862
Hynie S (1986) A simple procedure for the preparation of pertussis toxin and its use for the study of receptor-mediated inhibitory systems. Physiol Bohemoslov (in press)
Kostyuk PG (1984) Intracellular perfusion of nerve cells and its effects on membrane currents. Physiol Rev 64:435–454
Kurose H, Ui M (1983) Functional uncoupling of muscarinic receptors from adenylate cyclase in rat cardiac membranes by the active component of islet-activating protein, pertussis toxin. J Cyclic Nucleotide Protein Phosphor Res 9:305–318
Levy MN (1984) Cardiac sympathetic-parasympathetic interactions. Fed Proc 43:2598–2602
Löffelholz K, Pappano AJ (1985) The parasympathetic neuroeffector junction of the heart. Pharmacol Rev 37:1–24
Martin JM, Hunter DD, Nathanson NM (1985) Islet activating protein inhibits physiological responses evoked by cardiac muscarinic acetylcholine receptors. Role of guanosine triphosphate binding proteins in regulation of potassium permeability. Biochemistry 24:7521–7525
Moss J, Bruni P, Hsia JA, Tsai S-C, Watkins PA, Halpern JL, Burns DL, Kanaho Y, Chang PP, Hewlett EL, Vaughan M (1984) Pertussis toxin-catalyzed ADP-ribosylation: effects on the coupling of inhibitory receptors to the adenylate cyclase system. J Recept Res 4:459–474
Osterrieder W, Brum G, Hescheler J, Trautwein W, Flockerzi V, Hofmann F (1982) Injection of subunits of cyclic AMP-dependent protein kinase into cardiac myocytes modulates Ca2+ current. Nature 298:576–578
Pappano AJ, Biegon RL (1982) Mechanisms for muscarinic inhibition of calcium-dependent action potentials and contractions in developing ventricular muscle: the role of cyclic AMP. In: Paes de Carvalho AP, Hoffmann BF, Lieberman M (eds) Normal and abnormal conduction of the heartbeat. Futura Press, Mount Kisco, New York, pp 327–344
Pappano AJ, Inoue D (1984) Development of different electrophysiological mechanisms for muscarinic inhibitions of atria and ventricles. Fed Proc 43:2607–2612
Pappano AJ, Hartigan PM, Coutu MD (1982) Acetylcholine inhibits the positive inotropic effect of cholera toxin in ventricular muscle. Am J Physiol 243:H434-H441
Peterson GL (1977) A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem 83:346–356
Pfaffinger PJ, Martin JM, Hunter DD, Nathanson NM, Hille B (1985) GTP-binding proteins couple cardiac muscarinic receptors to a K channel. Nature 317:536–538
Reuter H (1983) Calcium channel modulation by neurotransmitters, enzymes and drugs. Nature 301:569–574
Sokolovsky M, Gurwitz D, Kloog J (1983) Biochemical characterization of the muscarinic receptors. Adv Enzymol 55:137–196
Sorota S, Tsuji Y, Tajima T, Pappano AJ (1985) Pertussis toxin treatment blocks hyperpolarization by muscarinic agonists in chick atrium. Circ Res 57:748–758
Sternweis PC, Robishaw JD (1984) Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. J Biol Chem 22:13806–13813
Trautwein W, Taniguchi J, Noma A (1982) The effect of intracellular cyclic nucleotides and calcium on the action potential and acetylcholine response of isolated cardiac cells. Pflügers Arch 392:307–314
Ui M, Katada T, Murayama T, Kurose H (1984) Selective blockage by islet-activating protein, pertussis toxin, of negative signal transduction from receptors to adenylate cyclase. In: Kito S, Segawa T, Kuriyama K, Yamamura HI, Olsen RW (eds) Neurotransmitter receptors — mechanism of action and regulation. Plenum Press, New York, pp 1–16
Watanabe M (1984) Biological activities of pertussigen from Bordetella pertussis of various agglutinogen types. Microbiol Immunol 28:509–515
Watanabe AM, Lindemann JP, Fleming JW (1984) Mechanisms of muscarinic modulation of protein phosphorylation in intact ventricles. Fed Proc 43:2618–2623
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Tuček, S., Doležal, V., Folbergrová, J. et al. Pertussis toxin inhibits negative inotropic and negative chronotropic muscarinic cholinergic effects on the heart. Pflugers Arch. 408, 167–172 (1987). https://doi.org/10.1007/BF00581347
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00581347
Key words
- Muscarinic effects on the heart
- Heart-blockade of cholinergic inhibition by pertussis toxin
- Pertussis toxin-blockade of cardiac muscarinic effects
- Guanine nucleotide binding proteins in the heart
- Cholinergic muscarinic receptors, coupling to channels
- Potassium channels, coupling to cardiac muscarinic receptors
- Calcium channels, coupling to cardiac muscarinic receptors
- Cyclic AMP, cholinergic receptors in the heart