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Inducible nitric oxide synthase in the myocard

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Abstract

Recognition of significance of nitric oxide synthases (NOS) in cardiovascular regulations has led to intensive research and development of therapies focused on NOS as potential therapeutic targets. However, the NOS isoform profile of cardiac tissue and subcellular localization of NOS isoforms remain a matter of debate. The aim of this study was to investigate the localization of an inducible NOS isoform (NOS2) in cardiomyocytes. Employing a novel immunocytochemical technique of a catalyzed reporter deposition system with tyramide and electron microscopical immunocytochemistry complemented with Western blotting and RT-PCR, we detected NOS2 both in rat neonatal and adult cultured cardiomyocytes and in the normal myocard of adult rats as well as in the human myocard of patients with dilative cardiomyopathy. NOS2 was targeted predominantly to a particulate component of the cardiomyocyte – along contractile fibers, in the plasma membrane including T-tubules, as well as in the nuclear envelope, mitochondria and Golgi complex. Our results point to an involvement of NOS2 in maintaining cardiac homeostasis and contradict to the notion that NOS2 is expressed in cardiac tissue only in response to various physiological and pathogenic factors. NOS2 targeting to mitochondria and contractile fibers suggests a relationship of NO with contractile function and energy production in the cardiac muscle.

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References

  1. Nathan C, Xie Q-W: Regulation of biosynthesis of nitric oxide. J Biol Chem 269: 13725–13728, 1994

    Google Scholar 

  2. Nathan C, Xie Q-W: Nitric oxide synthases: Roles, tolls, and controls. Cell 78: 915–918, 1994

    Google Scholar 

  3. Nicotera P, Brune B, Bagetta G: Nitric oxide: Inducer or suppressor of apoptosis. TiPS 18: 189–190, 1997

    Google Scholar 

  4. Förstermann U, Gath I, Schwarz P, Closs EI, Kleinert H: Isoforms of nitric oxide synthase: Properties, cellular distribution and expressional control. Biochem Pharmacol 50: 1321–1332, 1995

    Google Scholar 

  5. Moncada S, Higgs A, Furchgott R XIV: International Union of pharmacology nomenclature in nitric oxide research. Pharmacol Rev 49: 137–142, 1997

    Google Scholar 

  6. Hare JM, Colucci WS: Role of nitric oxide in the regulation of myocardial functions. Prog Cardiovasc Dis 38: 155–166, 1995

    Google Scholar 

  7. Singh K, Balligand JL, Fischer TA, Smith TW, Kelly RA: Glucocorticoids increase osteopontin expression in cardiac myocytes and microvascular endothelial cells. Role in regulation of inducible nitric oxide synthase. J Biol Chem 270: 28471–28478, 1995

    Google Scholar 

  8. Singh K, Balligand JL, Fischer TA, Smith TW, Kelly RA: Regulation of cytokine-inducible nitric oxide synthase in cardiac myocytes and microvascular endothelial cells. Role of extracellular signal-regulated kinases 1 and 2 (ERK1/ERK2) and STAT1 alpha. J Biol Chem 271: 1111–1117, 1996

    Google Scholar 

  9. Ungureanu-Longrois D, Balligand JL, Kelly RA, Smith TW: Myocardial contractile dysfunction in the systemic inflammatory response syndrome: Role of a cytokine-inducible nitric oxide synthase in cardiac myocytes. J Mol Cell Cardiol 27: 155–167, 1995

    Google Scholar 

  10. Wildhirt SM, Dudek RR, Suzuki H, Pito V, Narayan KS, Bing RJ: Immunocytochemistry in the identification of nitric oxide synthase isozymes in myocardial infarction. Cardiovasc Res 29: 526–531, 1995

    Google Scholar 

  11. Tsujino M, Hirata Y, Imai T, Kanno K, Eguchi S, Ito H, Marumo F: Induction of nitric oxide synthase gene by interleukin-1 in cultured rat cardiocytes. Circulation 90: 375–383, 1994

    Google Scholar 

  12. Oddis CV, Finkel MS: Cytokine-stimulated nitric oxide production inhibits mitochondrial activity in cardiac myocytes. Biochem Biophys Res Comm 213: 1002–1009, 1995

    Google Scholar 

  13. Buchwalow IB, Kostic MM, Schulze W, Wallukat G, Krause E-G, Haller H: Subcellular localization of inducible nitric oxide synthase in cardiomyocytes and control of its expression and activity. Circulation 96:(abstr) 938, 1997

    Google Scholar 

  14. Buchwalow IB, Schulze W, Kostic MM, Wallukat G, Morwinski R: Involvement of adenylyl cyclase in nitric oxide synthase expression by neonatal rat cardiomyocytes in vitro. Circulation 94: (abstr) 3451, 1996

    Google Scholar 

  15. Buchwalow IB, Schulze W, Kostic MM, Wallukat G, Morwinski R: Intracellular localization of inducible nitric oxide synthase in neonatal rat cardiomyocytes in culture. Acta Histochem 99: 231–240, 1997

    Google Scholar 

  16. Dusting GJ, MacDonald PS: Endogenous nitric oxide in cardiovascular disease and transplantation. Ann Med 27: 395–406, 1995

    Google Scholar 

  17. Russell ME, Wallace AF, Wyner LR, Newell JB, Karnovsky MJ: Upregulation and modulation of inducible nitric oxide synthase in rat cardiac allografts with chronic rejection and transplant arteriosclerosis. Circulation 92: 457–464, 1995

    Google Scholar 

  18. Shindo T, Ikeda U, Ohkawa F, Takahashi M, Funayama H, Nishinaga M, Kawahara Y, Yokoyama M, Kasahara T, Shimada K: Nitric oxide synthesis in rat cardiac myocytes and fibroblasts. Life Sci 55: 1101–1108, 1994

    Google Scholar 

  19. Shindo T, Ikeda U, Ohkawa F, Kawahara Y, Yokoyama M, Shimada K: Nitric oxide synthesis in cardiac myocytes and fibroblasts by inflammatory cytokines. Cardiovasc Res 29: 813–819, 1995

    Google Scholar 

  20. Nathan C: Inducible nitric oxide synthase: What difference does it make. J Clin Invest 100: 2417–2413, 1997

    Google Scholar 

  21. Halle W, Wollenberger A: Differentiation and behavior of isolated embryonic and neonatal heart cells in a chemically defined medium. Am J Cardiol 25: 292–299, 1970

    Google Scholar 

  22. Wallukat G, Wollenberger A: Supersensitivity to (β-adrenoceptor stimulation evoked in cultured neonatal rat heart myocytes by L(+)-lactate and pyruvate. J Auton Pharmacol 13: 1–14, 1993

    Google Scholar 

  23. Nees S, Dendorfer A: Regulation of the myocardial blood supply by adenosine and adenine nucleotides: Consequences of newly detected barrier functions of the coronary endothelium. In: G. Wilson, L. Illum, S.S. Davis (eds). Pharmaceutical Application of Cell and Tissue Culture. Plenum Press, New York/London, 1990, pp 72–79

    Google Scholar 

  24. Volz A, Piper HM, Siegmund B, Schwartz P: Longevity of adult ventricular rat heart muscle cells in serum-free primary culture. J Mol Cell Cardiol 23: 161–173, 1991

    Google Scholar 

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

    Google Scholar 

  26. Towbin T, Stahelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and applications. Proc Nat Acad Sci USA 76: 4350–4354, 1979

    Google Scholar 

  27. Schönfelder G, Mathias J, Hopp H, Fuhr N, van der Giet M, Paul M: Expression of inducible nitric oxide synthase in placenta of women with gestational diabetes. FASEB J 10: 777–784, 1996

    Google Scholar 

  28. Morwinski R, Karsten U, Wallukat G: Mast cells detected in cultures of neonatal rat heart cells. Agents Actions 41: 39–40, 1994

    Google Scholar 

  29. Hukkanen M, Hughes FJ, Buttery LD, Gross SS, Evans TJ, Seddon S, Riveros-Moreno V, Macintyre I, Polak JM: Cytokine-stimulated expression of inducible nitric oxide synthase by mouse, rat, and human osteoblast-like cells and its functional role in osteoblast metabolic activity. Endocrinology 136: 5445–5453, 1995

    Google Scholar 

  30. Weikert S, Freyer D, Weih M, Isaev N, Busch C, Schultze J, Megow D, Dirangl U: Rapid Ca2+ dependent NO production from central nervous system cells in culture measured by NO nitrite/ozone chemoluminescence. Brain Res 748: 1–11, 1997

    Google Scholar 

  31. Bia BL, Cassidy PJ, Young ME, Rafael JA, Leighton B, Davies KE, Radda GK, Clarke K: Decreased myocardial nNOS, increased iNOS and abnormal ECGs in mouse models of Duchenne muscular dystrophy. J Mol Cell Cardiol 31: 1857–1862, 1999

    Google Scholar 

  32. Rouet-Benzineb P, Eddahibi S, Raffestin B, Laplace M, Depond S, Adnot S, Crozatier B: Induction of cardiac nitric oxide synthase 2 in rats exposed to chronic hypoxia. J Mol Cell Cardiol 31: 1697–1708, 1999

    Google Scholar 

  33. De Belder AJ, Radomski MW, Why HJF, Richardson PJ, Bucknall CA, Salas E, Martin JF, Moncada S: Nitric synthase activities in human myocardium. Lancet 341: 84–85, 1993

    Google Scholar 

  34. Orus J, Heras M, Morales-Ruiz M, Leivas A, Roig E, Rigol M, Rivera F, Sanz G, Jimenez W: Nitric oxide synthase II mRNA expression in cardiac tissue of patients with heart failure undergoing cardiac transplantation. J Heart Lung Trans 19: 139–144, 2000

    Google Scholar 

  35. Moochalla S, Chatwall VJS, Chan STF, Ngoi SS, Chia YW, Rauff A: Nitric oxide synthase activity and expression in human colorectal cancer. Carcinogenesis 17: 1171–1174, 1996

    Google Scholar 

  36. Park CS, Park R, Krishna G: Constitutive expression and structural diversity of inducible isoform of nitric oxide synthase in human tissues. Life Sci 59: 219–225, 1996

    Google Scholar 

  37. Amin AR, Attur M, Vyas P, Leszczynskapiziak J, Levartovsky D, Rediske J, Clancy RM, Vora KA, Abramson SA: Expression of nitric oxide synthase in human peripheral blood mononuclear cells and neutrophils. J Inflammation 47: 190–205, 1996

    Google Scholar 

  38. Del Pozo V, de Arruda-Chaves E, de Andres B, Cardaba B, Lopez-Farre A, Gallardo S, Cortegano I, Vidarte L, Jurado A, Sastre J, Palomino P, Lahoz C: Eosinophils transcribe and translate messenger RNA for inducible nitric oxide synthase. J Immunol 158: 859–864, 1997

    Google Scholar 

  39. Wallerath T, Gath I, Aulitzky WE, Pollock JS, Kleinert H, Forstermann U: Identification of the NO synthase isoforms expressed in human neutrophyl granulocytes, megakaryocytes and platelets. Thromb Haemostasis 77: 163–167, 1997

    Google Scholar 

  40. Zackrisson U, Mikuni M, Wallin A, Delbro D, Hedin L, Brannstrom M: Cell specific localization of nitric oxide synthases (NOS) in the rat ovary during follicular development, ovulation and luteal formation. Hum Reprod 11: 2667–2673, 1996

    Google Scholar 

  41. Casado M, Diazguerra MJM, Rodrigo J, Fernandez AP, Bosca L, Martinsatz P: Expression of the calcium independent cytokine inducible (iNOS) isoform of nitric oxide synthase in rat placenta. Biochem J 324: 201–207, 1997

    Google Scholar 

  42. Miller MJS, Voelker CA, Olister S, Thompson JH, Zhang ZJ, Rivera D, Elobychildress S, Liu X, Clark DA, Pierce MR: Fetal growth retardation in rats may result form apoptosis: Role of peroxynitrite. Free Rad Biol Med 21: 619–629, 1996

    Google Scholar 

  43. Hoffman RA, Zhang GS, Nussler NC, Gleixner SL, Ford HR, Simmons RL, Watkins SC: Constitutive expression of inducible nitric oxide synthase in the mouse ileal mucosa. Am J Physiol Gastrointest Liver Physiol 35: 383–392, 1997

    Google Scholar 

  44. Gath I, Loss EI, Godtelarmbrust U, Schmitt S, Nakane M, Wessler I, Förstermann U: Inducible NO synthase II and neuronal NO synthase I are constitutively expressed in different structures of guinea pig skeletal muscle: Implications for contractile function. FASEB J 10: 1614–1620, 1996

    Google Scholar 

  45. Griffith OW, Stuehr DJ: Nitric oxide synthases: Properties and catalytic mechanism. Annu Rev Physiol 57: 707–736, 1995

    Google Scholar 

  46. Stuehr DJ, Cho HJ, Kwon NS, Weise MF, Nathan CF: Purification and characterization of the cytokine-induced macrophage nitric oxide synthase: An FAD-and FMN-containing flavoprotein. Proc Natl Acad Sci USA 88: 7773–7777, 1991

    Google Scholar 

  47. Kobzik L, Stringer B, Balligand JL, Reid MB, Stamler S: Endothelial type nitric oxide synthase in skeletal muscle fibers: Mitochondrial relationships. Biochem Biophys Res Comm 221: 375–381, 1995

    Google Scholar 

  48. Buchwalow IB, Kopiev OW, Schulze W: Ultracytochemical localization of adenylate cyclase in rat thymocytes. Histochemistry 72: 625–634, 1981

    Google Scholar 

  49. Schulze W, Buchwalow IB: Adenylyl cyclase in the heart: An enzymocytochemical and immunocytochemical approach. Micr Res Techn 40: 473–478, 1998

    Google Scholar 

  50. Schulze W, Buchwalow IB, Wolf W-P, Will-Shahab L: Comparative immunocytochemical localization of G proteins in the rat heart tissue. Acta Histochem 96: 87–95, 1994

    Google Scholar 

  51. Bates TE, Loesch A, Burnstock G, Clark GB: Mitochondrial nitric oxide synthase: A ubiquitous regulator of oxidative phosphorylation? Biochem Biophys Res Comm 218: 40–44, 1996

    Google Scholar 

  52. Ghafourifar P, Richter C: Nitric oxide synthase in mitochondria. FEBS Lett 418: 291–296, 1997

    Google Scholar 

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Authors and Affiliations

  1. Franz Volhard Clinic, Medical Faculty of the Charite, Humboldt University of Verlin, 13125, Berlin, Germany

    Igor B. Buchwalow

  2. Department of Medicine B, University of Münster, 48149, Münster, Germany

    Igor B. Buchwalow

  3. Max Delbrück Center for Molecular Medicine, 13125, Berlin, Germany

    Wolfgang Schulze, Peter Karczewski, Milosav M. Kostic, Gerd Wallukat, Rosmarie Morwinski & Ernst-Georg Krause

  4. German Cardiological Center, 13353, Berlin, Germany

    Johannes Müller

  5. Benjamin Franklin Medical Center, Institute for Clinical Pharmacology and Toxicology, Free University of Berlin, 14195, Berlin, Germany

    Martin Paul

  6. Slovak Academy of Sciences, Institute for Heart Research, Bratislava, Slovak Republic

    Jan Slezak

  7. Franz Volhard Clinic, Medical Faculty of the Charite, Humboldt University of Berlin, 13125, Berlin, Germany

    Friedrich C. Luft & Hermann Haller

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  1. Igor B. Buchwalow
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  2. Wolfgang Schulze
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  9. Martin Paul
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Correspondence to Igor B. Buchwalow.

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Buchwalow, I.B., Schulze, W., Karczewski, P. et al. Inducible nitric oxide synthase in the myocard. Mol Cell Biochem 217, 73–82 (2001). https://doi.org/10.1023/A:1007286602865

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