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Molecular and Cellular Biochemistry

, Volume 148, Issue 1, pp 89–94 | Cite as

Regulation of the calcium slow channel by cyclic GMP dependent protein kinase in chick heart cells

  • George E. Haddad
  • Nicholas Sperelakis
  • Ghassan Bkaily
Article

Abstract

In order to assess the interaction between the cAMP-dependent and the cGMP-dependent phosphorylation pathways on the slow Ca2+ current (ICa(L)), whole-cell voltage-clamp experiments were conducted on embryonic chick heart cells. Addition of 8Br-cGMP to the bath solution reduced the basal (unstimulated) ICa(L). Intracellular application of the catalytic subunit of PK-A (PK-A(cat); 1.5 μM) via the patch pipette rapidly potentiated ICa(L) by 215±16% (n=4); subsequent addition of 1 mM 8Br-cGMP to the bath reduced the amplitude of ICa(L) towards the initial control values (123±29%). Intracellular application of PK-G (25 nM pre-activated by 10−7 M cGMP), rapidly inhibited the basal ICa(L) by 64±6% (n=8). Heat-denatured PK-G was ineffective. Subsequent additions of relatively high concentrations of 8Br-cAMP (1 mM) or isoproterenol (ISO, 1–10 μM) did not significantly remove the PK-G blockade of ICa(L). The results of the present study suggest that: (a) 8Br-cGMP can inhibit the basal or stimulated (by PK-A(cat)) ICa(L) in embryonic chick myocardial cells. (b) PK-G applied intracellularly inhibits the basal ICa(L).

Key words

ICa(L) cyclic nucleotides PK-A PK-G isoproterenol embryo chick heart 

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References

  1. 1.
    Bean BP, Nowycky NC, Tsein RW: b-adrenergic modulation of calcium channels in frog ventricular heart cells. Nature 307: 371–375, 1984Google Scholar
  2. 2.
    Bkaily G, Sperelakis N: Injection of protein kinase inhibitor into cultured heart cells blocks calcium slow channels. Am J Physiol 246: H630-H634, 1984Google Scholar
  3. 3.
    Bkaily G, Sperelakis N: Injection of guanosine 5′-cyclic monophosphate into heart cells blocks calcium slow channels. Am J Physiol 248: H745-H749, 1985Google Scholar
  4. 4.
    Bkaily G: Single heart cells as models for studying cardiac toxicology. In: G. Jolles and A. Cordier (eds).In vitro methods in toxicology. Academic Press, London 1992, pp 289–334Google Scholar
  5. 5.
    Bkaily G, Perron N, Wang S, Sculptoreanu A, Jacques D, Menard D: Atrial natriuretic factor blocks the high-threshold Ca2+ current and increase K+ current in fetal single ventricular cells. J Mol Cell Cardiol 25: 1305–1316, 1993Google Scholar
  6. 6.
    Cachelin AB, dePeyer JE, Kokubun S, Reuter H: Ca2+ channel modulation by 8-bromo-cyclic AMP in culture heart cells. Nature 304: 462–464, 1983Google Scholar
  7. 7.
    Hartzell HC, Fischmeister R: Opposite effects of cyclic GMP and cyclic AMP on Ca2+ current in single heart cells. Nature 323: 273–275, 1986Google Scholar
  8. 8.
    Haase H, Karezewski P, Beckert R, Krause EG: Phosphorylation of the L-type calcium channel b subunit is involved in β-adrenergic signal transduction in canine myocardium. FEBS 335: 217–222, 1993Google Scholar
  9. 9.
    Mery PF, Lohmann SM, Walter U, Fischmeister R: Ca2+ current is regulated by cyclic GMP-dependent protein kinase in mammalian cardiac myocytes. Proc Natl Acad Sci 88: 1197–1201, 1991Google Scholar
  10. 10.
    Nargeot J, Nerbonne JM, Engels J, Lester HA: Time course of the increase in the myocardial slow inward current after a photochemically generated concentration jump of intracellular cAMP. Proc Natl Acad Sci 80: 2395–2399, 1983Google Scholar
  11. 11.
    Ono K, Fozzard HA: Phosphorylation restores activity of L-type calcium channels after rundown in inside-out patches from rabbit cardiac cells. J Physiol (London) 454: 673–688, 1992Google Scholar
  12. 12.
    Reuter H, Stevens CF, Tsein RW, Yellen G: Peptides of single calcium channels in cardiac cells culture. Nature 297: 501–504, 1982Google Scholar
  13. 13.
    Trautwein W, Hofmann F: Activation of calcium current by injection of cAMP and catalytic subunit of cAMP-dependent protein kinase. Proc Intern Union Physiol Sci 15: 75–83, 1983Google Scholar
  14. 14.
    Tsein RW, Giles W, Greengard P: Cyclic AMP mediates the action of adrenaline on the action potential plateau of cardiac Purkinje fibers. Nature 240: 181–183, 1972Google Scholar
  15. 15.
    Wahler GM, Rusch NJ, Sperelakis N: 8-Bromo-cyclic GMP inhibits the calcium channel current in embryonic chick ventricular myocytes. Can J Physiol Pharm 68: 531–534, 1990Google Scholar
  16. 16.
    Yue DT, Herzig S, Marban E: b-adrenergic stimulation of calcium channels occurs by potentiation of high-activity gating modes. Proc Natl Acad USA 87: 753–757, 1990Google Scholar
  17. 17.
    Levi RC, Alloatti G, Fischmeister R: Cyclic GMP regulates the Ca-channel current in guinea pig ventricular myocytes. Pflugers Arch 413: 685–687, 1989Google Scholar
  18. 18.
    Reuter H, Scholz H: The regulation of the calcium conductance of cardiac muscle by adrenaline. J Physiol (Lond) 264: 49–62, 1977Google Scholar
  19. 19.
    Ruth P, Rohrkasten, Biel M, Bosse E, Regulla S, Meyer HE, Flockerzi V, Hoffman F: Primary structure of the b subunit of the DHP-sensitive calcium channel from skeletal muscle. Science 245: 1115–1117, 1989Google Scholar
  20. 20.
    Shigenobu K, Sperelakis N: Calcium current channels induced by cathecholamines in chick embryonic hearts whose fast sodium channels are blocked by tetrodotoxin or elevated potassium. Circ Res 31: 932–952, 1972Google Scholar
  21. 21.
    Schneider J, Shigenobu K, Sperelakis N: Valinomycin inhibition of the inward slow current of cardiac muscle. In: P.E. Roy and N.S. Dhalla (eds). Recent Advances in Studies on Cardiac Structure and Metabolism. Baltimore, 1976, pp 33–52Google Scholar
  22. 22.
    Sperelakis N, Schneider J: A metabolic control mechanism for calcium ion influx that may protect the ventricular myocardial cell. Am J Cardiol 37: 1079–1085, 1976Google Scholar
  23. 23.
    Thakkar J, Tang SB, Sperelakis N, Wahler GM: Inhibition of cardiac slow action potentials by 8-bromo-cyclic GMP occurs independent of changes in cyclic AMP levels. Can J Physiol Pharmacol 66: 1092–1095, 1988Google Scholar
  24. 24.
    George WJ, Polson JB, O'Toole AG, Goldberg ND: Elevation of guanosine 3′,5′-cyclic phosphate in rat heart after perfusion with acetylcholine. Proc Natl Acad Sci 66: 398–403, 1970Google Scholar
  25. 25.
    Macleod KM, Diamond J: Effects of the cyclic GMP lowering agent LY83583 on the interaction of carbachol with forskolin in rabbit isolated cardiac preparations. J Pharmacol Exp Ther 238: 313–318, 1986Google Scholar
  26. 26.
    Watanabe AM, Green F, Ahmad Z: Studies on the cellular mechanisms of action of positive and negative inotropic agents. Basic Res Cardiol 84 suppl 1: 19–22, 1989Google Scholar
  27. 27.
    Godberg ND, Haddox MK, Nicol SE, Glass DB, Sanford CH, Kuehl Jr. FA, Estensen R: Biological regulation through opposing influences of cyclic GMP and cyclic AMP: The yin yang hypothesis. Adv Cyclic Nucl Res 5: 307–330, 1975Google Scholar
  28. 28.
    Wahler GM, Sperelakis N: Intracellular injection of cyclic GMP depresses cardiac slow action potentials. J Cyclic Nucl Prot Phos Res 10: 83–95, 1985Google Scholar
  29. 29.
    Tohse N, Sperelakis N: c-GMP inhibits the activity of single calcium channels in embryonic chick heart cells. Circ Res 69: 325–331, 1991Google Scholar
  30. 30.
    Ono K, Trautwein W: Potentiation by cyclic GMP of β-adrenergic effect on Ca2+ current in guinea-pig ventricular cells. J Physiol (London) 443: 387–404, 1991Google Scholar
  31. 31.
    Bkaily G, Payet MD, Benabderrazik M, Renaud J-F, Sauvé R, Bacaner MB, Sperelakis N; Bethanidine increased Na+ and Ca2+ currents and caused a positive inotropic effect in heart cells. Can J Physiol Pharmacol 66: 190–196, 1988Google Scholar
  32. 32.
    Bkaily G, Sculptoreanu A, Jacques D, Economos D, Ménard D: Apamin, a highly potent foetal L-type Ca2+ blocker in single heart cells. Am J Physiol 262: H463-H471, 1992Google Scholar
  33. 33.
    Fischmeister R, Hartzell HC: Cyclic guanosine 3′,5′-monophosphate regulates the calcium current in single cells from frog ventricle. J Physiol (London) 387: 455–472, 1987Google Scholar
  34. 34.
    Tritsh A, Hammond DC, Gershenfeld HM, Narin AC, Greengard P: cGMP-dependent protein kinase enhances Ca2+ current and potentiates the serotonin-induced Ca2+ current increase in snail neurons. Nature 323: 812–814, 1986Google Scholar
  35. 35.
    Klöckner U, Itagaki K, Bodi I, Schwartz A: β-subunit expression is required for cAMP-dependent increase of cloned cardiac and vascular calcium channel currents. Pflugers Arch 420: 413–415, 1992Google Scholar
  36. 36.
    Jahn H, Nastainczyk W, Rohrkasten A, Schneider T, Hoffmann F: Site-specific phosphorylation of the purified receptor for calciumchannel blockers by cAMP-and cGMP-protein kinases, protein kinase C, Calmodulin-dependent protein kinase II and Casein kinase II. Eur J Biochem 178: 535–542, 1988Google Scholar
  37. 37.
    Yoshida A, Takahashi M, Nishimura S, Takeshima H, Kokubun S: Cyclic AMP-dependent phosphorylation and regulation of the cardiac dihydropyridine-sensitive Ca channel. FEBS 309: 343–349, 1992Google Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • George E. Haddad
    • 1
    • 2
  • Nicholas Sperelakis
    • 1
  • Ghassan Bkaily
    • 2
  1. 1.Department of Physiology and Biophysics, College of MedicineUniversity of CincinnatiCincinattiUSA
  2. 2.Department of Physiology and Biophysics, Faculty of MedicineUniversity of SherbrookeSherbrookeCanada

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