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Alterations in inotropy, nitric oxide and cyclic GMP synthesis, protein phosphorylation and ADP-ribosylation in the endotoxin-treated rat myocardium and cardiomyocytes

  • Part II: Cardiac Hypertrophy and Failure
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Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

To evaluate the effects of the in vivo endotoxin treatment of the rat on (1) the contractile responses in the subsequently isolated papillary muscle to adrenergic and cholinergic agonists and (2) the biochemical parameters (cyclic GMP, nitric oxide synthesis, protein phosphorylation and ADP-ribosyslation) in the subsequently isolated cardiomyocytes. Following the in vivo endotoxin treatment (4 mg/kg i.p., 18 h), contractile responses to increasing amounts of isoprenaline or to increasing amounts of oxotremorine in the presence of a fixed amount of isoprenaline were determined in isolated papillary strips. Activities of nitric oxide synthase, guanylyl cyclase, as well as phosphorylation of phospholamban and troponin-inhibitory subunit, and pertussis toxin-catalyzed and endogenous ADP-ribosylations were determined in the intact cardiomyocytes and subcellular fractions. The increase in the force of contraction by isoprenaline was reduced, while its inhibition by oxotremorine was greater in the endotoxin-treated papillary strips. The activities of both nitric oxide synthase, primarily of the inducible form of the enzyme, and cytosolic guanylyl cyclase were higher while the phosphorylations of both phospholamban and troponin-inhibitory subunit were of lesser magnitude in the cardiomyocytes following the in vivo endotoxin treatment. Pertussis toxin-catalyzed ADP-ribosylation of the 41 kDa polypeptide, which is the alpha subunit of Gi, was also decreased. The results of the present study support the postulate that alterations in both the cyclic AMP and cyclic GMP signalling cascade contribute to the myocardial dysfunction caused by endotoxin and cytokines.

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References

  1. Levy MN, Martin PJ: Neural control of the heart. In: R. Berne (ed.) Handbook of Physiology Section 2, The cardiovascular system. Waverly Press, Bethesda, MD., 1979, pp 581–620

    Google Scholar 

  2. Sulakhe PV, Jagadeesh G, Braun AP: Cardiac autonomic receptors and adenylyl cyclase in health and disease. In: H. Rupp (ed.) Regulation of Heart Function: Basic Concepts and Clinical Applications. Georg Thieme Verlag, Stuttgart, 1986, pp 71–94

    Google Scholar 

  3. Watanabe AM: Cholinergic agonists and antagonists. In: M. Rosen, B.F. Hoffman (eds). Cardiac Therapy. Martinus Nijhoff, Boston, 1983, pp 95–144

    Google Scholar 

  4. Sulakhe PV, MacKay JE, Rokosh DG, Morris T, Phan TD: Parasympathic control of the heart: Subcellular mechanisms. In: N.S. Dhalla, G.N. Pierce, R.E. Beamish (eds). Heart Function and Metabolism. Martinus Nijhoff, Boston, 1987, pp 135–162

    Google Scholar 

  5. Osterrieder W, Brum G, Hescheler J, Trautwein W, Flockerzi V, Hofmann F: Injection of subunits of cyclic AMP-dependent protein kinase into cardiac myocytes modulates Ca2+ Current. Nature 298: 576–578, 1982

    PubMed  Google Scholar 

  6. Yatani A, Brown AM: Rapid beta-adrenergic modulation of cardiac calcium channel currents by a fast G protein pathway. Science 245: 71–74, 1989

    PubMed  Google Scholar 

  7. Yatani A, Imoto Y, Codina J, Hamilton SL, Brown AM, Birnbaumer L: The stimulatory G protein of adenylyl cyclase, Gs, also stimulates dihydropyridine-sensitive Ca2+ channels. Evidence for direct regulation independent of phosphorylation by CAMP-dependent protein kinase or stimulation by a dihydropyridine agonist. J Biol Chem 263: 9887–9895, 1988

    PubMed  Google Scholar 

  8. Sperelakis N, Wahler GM: Regulation of Ca2+ influx in myocardial cells by beta adrenergic receptors, cyclic nucleotides, and phosphorylation. Mol Cell Biochem 82: 19–28, 1988

    Article  PubMed  Google Scholar 

  9. Lindemann JP, Jones LR, Hathaway DR, Henry BG, Watanabe AM: Beta-adrenergic stimulation of phospholamban phosphorylation and Ca2+-ATPase activity in guinea pig ventricles. J Biol Chem 258: 464–471, 1983

    PubMed  Google Scholar 

  10. Tada M, Katz AM. Phosphorylation of the sarcoplasmic reticulum and sarcolemma. Annu Rev Physiol 44: 401–423, 1982

    Article  PubMed  Google Scholar 

  11. England PJ. Phosphorylation of cardiac muscle In: A.J. Drake-Holland, M.I.M. Noble (eds). Cardiac Metabolism. John Wiley and Sons, U.K., 1983, pp 365–389

    Google Scholar 

  12. Solaro RJ. Overview of the role of Ca2+ and protein phosphorylation in contraction of the heart. In: R.J. Solaro (ed.) Protein Phosphorylation in Heart Muscle. CRC Press, Boca Raton, 1986, pp 1–15

    Google Scholar 

  13. 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, 1970

    PubMed  Google Scholar 

  14. Beavo JA, Conti M, Heaslip RJ: Multiple cyclic nucleotide phosphodiesterases. Mol Pharmacol 46: 399–405, 1994

    PubMed  Google Scholar 

  15. Kirstein M, Rivet-Bastide M, Hatem S, Benardeau A, Mercadier JJ, Fischmeister R. Nitric oxide regulates the calcium current in isolated human atrial myocytes. J Clin Invest 95: 794–802, 1995

    PubMed  Google Scholar 

  16. Levi RC, Alloatti G, Penna C, Gallo MP: Guanylate-cyclase-mediated inhibition of cardiac ICa by carbachol and sodium nitroprusside. Pflugers Arch 426: 419–426, 1994

    PubMed  Google Scholar 

  17. Mery PF, Pavoine C, Belhassen L, Pecker F, Fischmeister, R: Nitric oxide regulates cardiac Ca2+ current. Involvement of cGMP-inhibited and cGMP-stimulated phosphodiesterases through guanylyl cyclase activation. J Biol Chem 268: 26286–26295, 1993

    PubMed  Google Scholar 

  18. 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, 1991

    PubMed  Google Scholar 

  19. Shah AM, Spurgeon HA, Sollott SJ, Talo A, Lakatta EG: 8-bromocGMP reduces the myofilament response to Ca2+ in intact cardiac myocytes. Circ Res 74: 970–978, 1994

    PubMed  Google Scholar 

  20. Sulakhe PV, Sulakhe SJ, Leung NLK, St. Louis PJ, Hickie, RA: Guanylate cyclase: Subcellular distribution in cardiac muscle, skeletal muscle, cerebral cortex and liver. Biochem J 157: 705–712, 1976

    PubMed  Google Scholar 

  21. Sulakhe SJ, Leung NLK, Sulakhe PV: Properties of particulate, membrane-associated and soluble guanylate cyclase from cardiac muscle, skeletal muscle, cerebral cortex and liver. Biochem J 157: 713–719, 1976

    PubMed  Google Scholar 

  22. St. Louis PJ, Sulakhe PV: Adenylate cyclase, guanylate cyclase and cyclic nucleotide phosphodiesterases of guinea-pig cardiac sarcolemma. Biochem J 158: 535–541, 1976

    PubMed  Google Scholar 

  23. Narayanan N, Sulakhe PV: Magnesium- and manganese-supported guanylate cyclase in guinea-pig heart: Subcellular distribution and some properties of the microsomal enzyme. Int J Biochem 13: 1133–1141, 1981

    Article  PubMed  Google Scholar 

  24. Murad F: Regulation of cytosolic guanylyl cyclase by nitric oxide: The NO-cGMP signal transduction system. In: F. Murad (ed). Cyclic GMP: Synthesis, Metabolism, and Function. Advances in Pharmacology Vol. 26, Academic Press, San Diego, 1994, pp 19–30

    Google Scholar 

  25. Hare JM, Keaney JF, Balligand JL, Loscalzo J, Smith TW: Role of nitric oxide in parasympathetic modulation of β-adrenergic myocardial contractility in normal dogs. J Clin Invest 95: 360–366, 1995

    PubMed  Google Scholar 

  26. Schmidt HHHW, Lohmann SM, Walter U: The nitric oxide and cGMP signal transduction system: Regulation and mechanisms of action. Biochim Biophys Acta 1178: 153–175, 1993

    Article  PubMed  Google Scholar 

  27. Balligand JL, Kelly RA, Marsden PA, Smith TW, Michel T: Control of cardiac muscle cell function by an endogenous nitric oxide signaling system. Proc Natl Acad Sci 90: 347–351, 1993

    PubMed  Google Scholar 

  28. Han X, Shimoni Y, Giles WR: An obligatory role for nitric oxide in autonomic control of mammalian heart rate. J Physiol (Lond) 476: 309–314, 1994

    Google Scholar 

  29. Balligand JL, Ungureanu-Longrois D, Simmons WW, Pimental D, Malinski TA, Kapturczak M, Taha Z, Lowenstein CJ, Davidoff AJ, Kelly RA, Smith TW, Michel T: Cytokine-inducible nitric oxide syn thase (iNOS) expression in cardiac myocytes. Characterization and regulation of iNOS expression and detection of iNOS activity in single cardiac myocytes in vitro. J Biol Chem 269: 27580–27588, 1994

    PubMed  Google Scholar 

  30. Balligand JL, Ungureanu D, Kelly RA, Kobzik L, Pimental D, Michel T, Smith TW: Abnormal contractile function due to induction of nitric oxide synthesis in rat cardiac myocytes follows exposure to activated macrophage-conditioned medium. J Clin Invest 91: 2314–2319, 1993

    PubMed  Google Scholar 

  31. Brady AJB, Poole-Wilson PA, Harding SE, Warren JB: Nitric oxide production within cardiac myocytes reduces their contractility in endotoxemia. Am J Physiol 263: H1963-H1966, 1992

    Google Scholar 

  32. Schulz R, Panas DL, Catena R, Moncada S, Olley PM and Lopaschuk GD: The Role of nitric oxide in cardiac depression induced by interleukin-1 β and tumour necrosis factor-α. Br J Pharmacol 107: 27–34, 1994

    Google Scholar 

  33. Parker JL, Adams HR: Development of myocardial dysfunction in endotoxic shock. Am J Physiol 248: H818-H826, 1985

    PubMed  Google Scholar 

  34. Suffredini AF, Fromm RE, Parker MM, Brenner M, Kovacs JA, Wesley RA, Parrillo JE: The cardiovascular response of normal humans to administration of endotoxin. New Eng J Med 321: 280–287, 1989

    PubMed  Google Scholar 

  35. Schulz R, Triggle CR: Role of nitric oxide in vascular smooth muscle and cardiac muscle function. TIPS 15: 255–259, 1994

    PubMed  Google Scholar 

  36. Gulick T, Chung MK, Pieper SJ, Lange LG, Schreiner GF: Interleukinland tumor necrosis factor inhibit cardiomyocyte β-adrenergic responsiveness. Proc Nat] Acad Sci 86: 6753–6757, 1989

    Google Scholar 

  37. Piper HM, Volz A, Schwartz P: Adult ventricular rat heart muscle cells. In: H.M. Piper (ed). Cell Culture Techniques in Heart and Vessel Research. Springer-Verlag, Berlin, 1990, pp 36–60

    Google Scholar 

  38. Sulakhe PV, Vo XT: Regulation of phospholamban and Troponin-I phosphorylation in the intact rat cardiomyocytes by adrenergic and cholinergic stimuli: Roles of cyclic nucleotides, calcium, protein kinases and phosphatases and depolarization. Mol Cell Biochem 149/150: 103–126, 1995

    Google Scholar 

  39. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970

    PubMed  Google Scholar 

  40. Brune B, Lapetina EG: Activation of a cytosolic ADP-ribosyltransferase by nitric oxide generating agents. J Biol Chem 264: 8455–8458, 1989

    PubMed  Google Scholar 

  41. Braun AP, Gupta RC, Sulakhe PV: Pertussis toxin-sensitive G-proteins in the sino-atrial node and right atrium of bovine heart. Eur J Pharmacol (Mol Pharmacol Section) 189: 105–109, 1990

    Google Scholar 

  42. Schulz R, Nava E, Moncada, S: Induction and potential biological relevance of a Ca2+-independent nitric oxide synthase in the myocardium. Br J Pharmacol 105: 575–580, 1992

    PubMed  Google Scholar 

  43. Tao S. McKenna TM: In Vitro Endotoxin exposure induces contractile dysfunction n adult rat cardiac myocytes. Am J Physiol 267: H1745-H1752, 1994

    PubMed  Google Scholar 

  44. Evans HG, Lewis MJ Shah AM: Interleukin-1β Modulates myocardial contraction via dexamethasone sensitive production of nitric oxide. Cardiovasc Res 27: 1486–1490, 1993

    PubMed  Google Scholar 

  45. Weyrich AS, Ma XL, Buerke M, Murohara T, Armstead VE, Lefer AM, Nicolas JM, Thomas AP, Lefer DJ, Vinten-Johansen J: Physiological concentrations of nitric oxide do not elicit an acute negative inotropic effect in unstimulated cardiac muscle. Circ Res 75: 692–700, 1994

    PubMed  Google Scholar 

  46. Stein B, Drogemuller A, Mulsch A, Schmitz W. Scholz H: Ca++-dependent constitutive nitric oxide synthase is not involved in the cyclic GMP-increasing effects of carbachol in ventricular cardiomyocytes. J Pharmacol Exp Ther 266: 919–925, 1993

    PubMed  Google Scholar 

  47. Wahler GM, Dollinger SJ: Nitric oxide donor SIN-1 inhibits mammalian cardiac calcium current through cGMP-dependent protein kinase Am J Physiol 268: C45-C54, 1995

    PubMed  Google Scholar 

  48. Rozanski G, Witt R: IL-1 inhibits β-adrenergic control of cardiac calcium current: Role of L-arginine/nitric oxide pathway. Am J Physiol 267: H1753-H1758, 1994

    PubMed  Google Scholar 

  49. Coyler J: Control of the calcium pump of cardiac sarcoplasmic reticulum. A specific role for the pentameric structure of phospholamban? Cardiovasc Res 27: 1766–1771, 1993

    PubMed  Google Scholar 

  50. James PM, Inui M, Tada M, Chiesi M, Carafoli E: Nature and site of phospholamban regulation of the Ca2+ pump of sarcoplasmic reticulum. Nature 342: 92–95, 1989

    PubMed  Google Scholar 

  51. Kirchberger MA, Tada M, Katz M: Adenosine 3′:5′-monophosphatedependent protein kinase-catalyzed phosphorylation reaction and its relationship to calcium transport in cardiac sarcoplasmic reticulum. J Biol Chem 249: 6166–6173, 1974

    PubMed  Google Scholar 

  52. Tada M, Kirchberger MA, Repke DI, Katz AM: The stimulation of calcium transport in cardiac sarcoplasmic reticulum by adenosine 3′:5′-monophosphate-dependent protein kinase. J Biol Chem 249: 6174–6180, 1974

    PubMed  Google Scholar 

  53. Reithmann C, Giershik P, Werdan K, Jakobs KH: Tumor necrosis factor-α up-regulates Gia and GB proteins and adenylyl cyclase responsiveness in rat cardiomyocytes. Eur J Pharmacol (Mol Pharmacol Section) 206: 53–60, 1991

    Google Scholar 

  54. Brune B, Dimmeler S, Vedia LM, Lapetina EG: Nitric oxide: A signal for ADP-ribosylation of proteins. Life Sci 54: 61–70, 1993

    Google Scholar 

  55. Dimmeler S, Lottspeich F, Brune B: Nitric oxide causes ADPribosylation and inhibition of glyceraldehyde-3-phosphate dehydro-genase. J Biol Chem 267: 16771–16774, 1992

    PubMed  Google Scholar 

  56. Zhang J, Snyder SH: Purification of a nitric oxide-stimulated ADPribosylated protein using biotinylated β-nicotinamide adenine dinucleotide. Biochemistry 32: 2228–2233, 1993

    PubMed  Google Scholar 

  57. Piron KJ, McMahon KK: Localization and partial characterization of ADP-ribosylation products in hearts from adult and neonatal rats. Biochem J 270: 591–597, 1990

    PubMed  Google Scholar 

  58. Quist EE, Coyle DL, Vasan R, Satumtira N, Jacobson EL, Jacobson MK: Modification of cardiac membrane adenylyl cyclase activity and Gsα by NAD and endogenous ADP-ribosyltransferase. J Mol Cell Cardiol 26: 251–260, 1994

    PubMed  Google Scholar 

  59. McDonald LJ, Moss J: Stimulation by nitric oxide of a novel linkage of NAD to Glyceraldehyde-3-phosphate dehydrogenase. Proc Natl Acad Sci 90: 6238–6241, 1993

    PubMed  Google Scholar 

  60. McDonald LJ, Moss J: Nitric oxide and NAD-dependent protein modification. Mol Cell Biochem 138: 201–206, 1994

    PubMed  Google Scholar 

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Author notes

  1. Willard J. Costain

    Present address: Department of Pharmacology, Faculty of Medicine, McMaster University, Hamilton, Ontario, Canada

Authors and Affiliations

  1. Department of Physiology, College of Medicine, University of Saskatchewan, 107 Wiggins Road, S7N 5E5, Saskatoon, Canada

    Prakash V. Sulakhe, Lakshman Sandirasegarane, J. Paul Davis, Xuan T. Vo, Willard J. Costain & Rahul R. Mainra

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  2. Lakshman Sandirasegarane
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Sulakhe, P.V., Sandirasegarane, L., Davis, J.P. et al. Alterations in inotropy, nitric oxide and cyclic GMP synthesis, protein phosphorylation and ADP-ribosylation in the endotoxin-treated rat myocardium and cardiomyocytes. Mol Cell Biochem 163, 305–318 (1996). https://doi.org/10.1007/BF00408671

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