Pflügers Archiv

, Volume 392, Issue 4, pp 307–314 | Cite as

The effect of intracellular cyclic nucleotides and calcium on the action potential and acetylcholine response of isolated cardiac cells

  • W. Trautwein
  • J. Taniguchi
  • A. Noma
Excitable Tissues and Central Nervous Physiology

Abstract

Ventricular and atrioventricular nodal cells from guinea pig and rabbit hearts were isolated by perfusing the heart with collagenase (Langendorff perfusion). In these cells the cyclic nucleotides cAMP and cGMP or Ca and EGTA were injected through a microelectrode by pressure (0.5–3 kg/cm2). The effect of injection on both the action potential and the hyperpolarization induced by acetylcholine was studied. The following results were obtained.
  1. 1.

    cAMP prolonged the ventricular action potential and shifted the plateau to more positive potentials. The configuration of the A-V nodal action potential was not detectably changed by cAMP injection, but the spontaneous rate was increased.

     
  2. 2.

    cGMP first shortened the ventricular action potential. In most experiments this effect was followed by long lasting prolongation of the action potential.

     
  3. 3.

    Both extracellular and intracellular application of dibutyryl cGMP shortened the ventricular action potential but did not produce a subsequent prolongation. However, prolongation was observed on injection of GMP, the direct metabolite.

     
  4. 4.

    Injection of cGMP in nodal cells did not hyperpolarize the membrane nor slow the spontaneous rate; rather, an increase in rate was observed.

     
  5. 5.

    The acetylcholine-induced hyperpolarization was not altered in amplitude or time course by the injection of cAMP, cGMP, Ca or EGTA.

     
  6. 6.

    The results support the hypothesis that cGMP might be involved in the control of voltage-controlled ionic channels but suggest that it does not play a role as a mediator of the classical muscarinic action i.e. the activation of a specific potassium channel by acetylcholine.

     

Key words

c-AMP, c-GMP Injection A-V nodal cell Ventricular cell Dibutyryl c-GMP GMP 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Beresewicz A, Reuter H (1977) The effects of adrenaline and theophilline on action potential and contraction of mammalian ventricular muscle under “rested-state” and “steady-state” stimulation. Naunyn Schmiedeberg's Arch Pharmacol 301: 99–107Google Scholar
  2. Brooker G (1977) Dissociation of cyclic GMP from the negative inotropic action of carbachol in guinea pig atria. J Cyclic Nucl Res 3: 407–413Google Scholar
  3. Carmeliet E, Ramon J (1980) Effects of acetylcholine on the time-dependent currents in sheep cardiac Purkinje fibers. Pflügers Arch 387: 207–215Google Scholar
  4. Diamond J, Ten Eick RT, Trapani AJ (1977) Are increases in cyclic GMP levels responsible for the negative inotropic effects of acetylcholine in the heart? Biochem Biophys Res Comm 79: 912–918Google Scholar
  5. Di Francesco D, Noma A, Trautwein W (1980) Separation of current induced by potassium accumulation from acetylcholine-induced relaxation current in the rabbit S-A node. Pflügers Arch 387: 83–90Google Scholar
  6. Drummond GI, Severson DL (1979) Cyclic nucleotides and cardiac function. Circ Res 44: 145–153Google Scholar
  7. Drummond GI, Hemmings S, Warneboldt RB (1974) Uptake and catabolism of N6,2′-0-dibutyryl cyclic AMP by the perfused heart. Life Sci 15: 319–328Google Scholar
  8. Flitney FW, Singh J (1980) Depressant effect of 8-bromo guanosine 3′,5′-cyclic monophosphate on endogeneous adenosine 3′,5′-cyclic monophosphate levels in frog ventricle. J Physiol (Lond) 302: 29P-30PGoogle Scholar
  9. George WJ, Wilkerson RD, Kadowitz PJ (1973) Influence of acetylcholine on contractile force and cyclic nucleotide levels in the isolated perfused rat heart. J Pharmacol Exp Ther 184: 228–235Google Scholar
  10. Giles WR, Noble SJ (1976) Changes in membrane currents in bull-frog atrium produced by acetylcholine. J Physiol (Lond) 261: 103–123Google Scholar
  11. Glitsch HG, Pott L (1978) Effects of acetylcholine in parasympathic nerve stimulation on membrane potential in quiescent guinea-pig atria. J Physiol (Lond) 279: 655–668Google Scholar
  12. Harris EJ, Hutter OH (1956) The action of acetylcholine on the movements of potassium ions in the sinus venosus of the heart. J Physiol (Lond) 133: 58PGoogle Scholar
  13. Hino N, Ochi R (1980) Effect of acetylcholine on membrane currents in guinea-pig papillary muscle. J Physiol (Lond) 307: 183–197Google Scholar
  14. Hoffman BF, Cranefield PF (1960) Electrophysiology of the heart. McGraw Hill, New YorkGoogle Scholar
  15. Ikemoto Y, Goto M (1975) Nature of the negative inotropic effect of acetylcholine on the myocardium. An elucidation of the bullfrog atrium. Proc Jpn Acad 51: 501–505Google Scholar
  16. Isenberg G (1977a) Cardiac Purkinje fibers. Resting, action, and pacemaker potential under the influence of [Ca]i as modified by intracellular injection technique. Pflügers Arch 371: 51–59Google Scholar
  17. Isenberg G (1977b) Cardiac Purkinje fibers. The slow inward current component under the influence of modified [Ca]i. Pflügers Arch 371: 61–69Google Scholar
  18. Isenberg G (1977c) Cardiac Purkinje fibers [Ca]i controls steady state potassium conductance. Pflügers Arch 371: 71–76Google Scholar
  19. Isenberg G (1977d) Cardiac Purkinje fibers. [Ca]i controls the potassium permeability via the conductance componentsg K1 andg K2. Pflügers Arch 371: 77–85Google Scholar
  20. Kehoe JS, Marty A (1980) Certain slow synaptic responses: their properties and possible underlying mechanisms. Ann Rev Biophys Bioeng 9: 437–460Google Scholar
  21. Kohlhardt M, Haap K (1978) 8-Bromo-guanosine-3′,5′-monophosphate mimics the effect of acetylcholine on slow response action potential and contractile force in mammalian atrial myocardium. J Mol Cell Cardiol 10: 573–586Google Scholar
  22. Mirro MJ, Bailey JC, Watanabe AM (1979) Dissociation between the electrophysiological properties and total tissue cyclic guanosine monophosphate content of guinea pig atria. Circ Res 45: 225–233Google Scholar
  23. Moustafa E, Skomedal T, Osnes JB, Øye I (1976) Cyclic AMP formation and morphology of myocardial cells isolated from adult heart: Effect of Ca2+ and Mg2+. Biochim Biophys Acta 421: 411–415Google Scholar
  24. Nawrath H (1977) Does cyclic GMP mediate the negative inotropic effect of acetylcholine in the heart? Nature 267: 72–74Google Scholar
  25. Niedergerke R, Page S (1977) Analysis of catecholamine effects in single atrial trabeculae of the frog heart. Proc R Soc B 197: 33–362Google Scholar
  26. Noma A, Trautwein W (1978) Relaxation of the ACh-induced potassium current in the rabbit sinoatrial node cell. Pflügers Arch 377: 193–200Google Scholar
  27. Noma A, Peper K, Trautwein W (1979a) Acetylcholine-induced potassium current fluctuations in the rabbit sino-atrial node. Pflügers Arch 381: 263–269Google Scholar
  28. Noma A, Osterrieder W, Trautwein W (1979b) The effect of external potassium on the elementary conductance of the ACh-induced potassium channel in the sino-atrial node. Pflügers Arch 381: 263–269Google Scholar
  29. Osterrieder W, Noma A, Trautwein W (1980) On the kinetics of the potassium channel activated by acetylcholine in the S-A node of the rabbit heart. Pflügers Arch 386: 101–109Google Scholar
  30. Pelzer D, Trautwein W (1981) Zum Mechanismus der negativ inotropen Acetylcholin (ACh)-Wirkung auf das Ventrikelmyokard. German J Cardiol 70: 308: R 202Google Scholar
  31. Powell T, Twist VW (1976) Isoprenaline stimulation of cyclic AMP production by isolated cells from adult rat myocardium. Biochem Biophys Res Commun Chem Pathol Pharmacol 72: 1218–1225Google Scholar
  32. Powell T, Terrar DA, Twist VW (1978) Electrical activity in superfused cells isolated from adult rat ventricular myocardium. J Physiol (Lond) 284: 148 PGoogle Scholar
  33. Reuter H (1974) Localization of beta adrenergic receptors and effects of noradrenaline and cyclic nucleotides on action potentials, ionic currents and tension in mammalian cardiac muscle. J Physiol (Lond) 242: 429–451Google Scholar
  34. Reuter H (1979) Properties of two inward membrane currents in the heart. Ann Rev Physiol 41: 413–424Google Scholar
  35. Reuter H, Scholz H (1977) The regulation of the Ca conductance of cardiac muscle by adrenaline. J Physiol (Lond) 264: 49–62Google Scholar
  36. Taniguchi J, Kokubun S, Noma A, Irisawa H (1981) Spontaneously active cells isolated from the sino-atrial and atrio-ventricular nodes of the rabbit heart. Jpn J Physiol 31: 547–558Google Scholar
  37. Ten Eick R, Nawrath H, McDonald TF, Trautwein W (1976) On the mechanism of negative inotropic effect of acetylcholine. Pflügers Arch 361: 207–213Google Scholar
  38. Tsien RW (1973) Adrenaline-like effects of intracellular iontophoresis of cyclic AMP in cardiac Purkinje fibres. Nature 245: 120–122Google Scholar
  39. Tsien RW, Giles W, Greengard P (1972) Cyclic AMP mediates the effects of adrenaline on cardiac Purkinje fibres. Nature 240: 1821–1823Google Scholar
  40. Trautwein W (1963) Generation and conduction fo impulse in the heart as affected by drugs. Pharmacol Rev 15: 277–332Google Scholar
  41. Trautwein W, Dudel J (1958) Zum Mechanismus der Membranwirkung des Acetylcholins an der Herzmuskelfaser. Pflügers Arch ges Physiol 266: 324–334Google Scholar
  42. Watanabe AM, Besch HR Jr (1975) Interaction between cyclic adenosine monophosphate and cyclic guanosine monophosphate in guinea pig ventricular myocardium. Circ Res 37: 309–317Google Scholar
  43. Watanabe AM, McConnaughey MM, Strawbridge RA, Fleming JW, Jones LR, Besch HR Jr (1978) Muscarinic cholinergic receptor modulation of β-adrenergic receptor affinity for catecholamines. J Biol Chem 253: 4833–4836Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • W. Trautwein
    • 1
  • J. Taniguchi
    • 1
  • A. Noma
    • 1
  1. 1.National Institute for Physiological SciencesOkazakiJapan

Personalised recommendations