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Myocardial Ca-sequestration failure and compensatory increase in Ca-ATPase with congestive cardiomyopathy: kinetic characterization by a homogenate microassay using real-time ratiometric indo-1 spectrofluorometry

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Abstract

A novel, simple, rapid and reproducible microassay is used for kinetic analysis of Ca-sequestration by homogenates of myocardium of turkeys with furazolidone-induced congestive cardiomyopathy. The assay monitors Ca in real-time using dual-emission ratiometric spectrofluorometry and the Ca-indicator dye indo-1. Using this assay and isolated SR studies we make several novel findings regarding the mechanism of SR failure in furazolidone cardiomyopathy.

Qualitative differences in Ca-sequestration were not detected between groups. However, compared to controls the furazolidone treatment resulted in: 1) 50% depression in maximal activities (1.54 ± 0.36 vs 0.73 ± 0.12 µM/sec); 2) 2-fold increases in post-sequestration concentrations of ionized Ca(79 ± 23 vs 141 ±13 nmol Ca/L homogenate); 3) 2-fold increases in Ca half-life (415 vs 790 msec); and 4) 25% increased passive Ca-binding capacity of homogenates. The Ca-ATPase specific activity of isolated sarcoplasmic reticulum was 60% increased in congestive cardiomyopathy (543 ± 140 vs 873 ± 108 nmol ATP hydrolyzed/min/mg membrane protein) although membrane yield was 20% decreased (0.79 ± 0.09 vs 0.63 ± 0.03 mg/g heart). The increased ATPase and decreased Ca-uptake activities in combination with the occurrence of 36% cardiac hypertrophy and 19% decreased body weights resulted in estimates of the relative energy cost to the animal for myocardial Ca transport being 5.5-fold increased with cardiomyopathy (20.5 vs 111 nmol ATP hydrolyzed per µM decrease of sarcoplasmic free Ca/kg body weight).

These data indicate that congestive cardiomyopathy is associated with markedly increased permeability of sarcoplasmic reticulum to Ca and compensatorily increased Ca-ATPase activity. Accelerated energy consumption due to the increased energy cost of Ca transport and increased time of myocyte activation are predicted to predispose the myocardium to fatigue and irreversible failure.

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References

  1. O'Brien PJ: Calcium sequestration by isolated sarcoplasmic reticulum: real-time monitoring using ratiometric dual-emission spectrofluorometry and the fluorescence calcium-binding dye indo-1. Molec Cell Biochem 94: 113–119, 1990

    Google Scholar 

  2. O'Brien PJ: Porcine malignant hyperthermia susceptibility: increased calcium sequestering activity of skeletal muscle sarcoplasmic reticulum. Can J Vet Res 51: 50–55, 1986

    Google Scholar 

  3. O'Brien PJ, Ianuzzo CD, Moe GW, Stopps TP, Armstrong PW: Rapid ventricular pacing of dogs to heart failure: biochemical and physiological studies. Can J Physiol Pharmacol 68:34–39, 1990

    Google Scholar 

  4. O'Brien PJ, Ianuzzo CD, Wittnich C, Moe GW, Armstrong PW. Cardiac and muscle fatigue due to relative functional overload induced by tachycardia, hypersensitivity, or ischemia correlates with sarcoplasmic reticulum failure. Can J Physiol Pharmacol, In press

  5. Rapundalo ST, Briggs NF, Feher JJ: Effects of ischemia on the isolation and function of canine cardiac sarcoplasmic reticulum. J Mol Cell Cardiol 18: 837–851, 1986

    Google Scholar 

  6. Mirsalimi SM, Quereshi FS, Julian RJ, O'Brien PJ: Myocardial biochemical changes in furazolidone-induced cardiomyopathy of turkeys. J Comp Path 102: 139–147, 1990

    Google Scholar 

  7. Haryigaya S, Schwartz A: Rate of calcium binding and uptake in normal animal and failing human cardiac muscle. Circ Res 25: 781–794, 1969

    Google Scholar 

  8. O'Brien PJ, Ling E, Williams HM, Brotherton S, Salerno T, Lumsden JH, Ianuzzo CD: Compensatory adaptation of the heart to chronic rate overload: increase in calcium transport ATPase activity of myocardial sarcoplasmic reticulum. Can J Cardiol 4: 243–250, 1988

    Google Scholar 

  9. Grynkiewicz G, Poenie M, Tsien RY: A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem 260: 3440–3450, 1985

    CAS  PubMed  Google Scholar 

  10. Gertz EW, Hess ML, Lain RF, Briggs FN: Activity of the vesicular calcium pump in the spontaneously failing heart-lung preparation. Circ Res 20: 477–484, 1967

    Google Scholar 

  11. Lee KS, Ladinsky H, Stuckey JH: Decreased Ca2+ uptake by sarcoplasmic reticulum after coronary artery occlusion for 60 and 90 minutes. Circ Res 21: 439–443, 1967

    Google Scholar 

  12. Feher JJ, LeBolt WR, Manson NH: Differential effect of global ischemia on the ryanodine-sensitive and insensitive calcium uptake of cardiac sarcoplasmic reticulum. Circ Res 65: 1400–1408, 1989

    Google Scholar 

  13. Lecarpentier Y, Martin JL, Gastineau P, Hatt PY: Load dependence of mammalian heart relaxation during cardiac hypertrophy and heart failure. Am J Physiol 242: H855-H861, 1982

    Google Scholar 

  14. Lindenmayer GE, Sordahl LA, Harigaya S, Allen JC, Besch H, Schwartz A: Some biochemical studies on subcellular systems isolated from fresh recipient human cardiac tissue obtained during transplantation. Am J Cardiol 27: 277–283, 1971

    Google Scholar 

  15. Muir JR, Dhalla NS, Orteza JM, Olson RE: Energy-linked calcium transport in subcellular fractions of the failing rat heart. Circ Res 26: 429–438, 1970

    Google Scholar 

  16. Nayler WG: The role of calcium in the ischemic myocardium. Am J Pathol 102: 262–270, 1981

    Google Scholar 

  17. Suko J, Vogel JHK, Chidsey CA: Intracellular calcium and myocardial contractility. III. Reduced calcium uptake and ATPase of the sarcoplasmic reticular fraction prepared from chronically failing calf hearts. Circ Res 27: 235–247, 1970

    Google Scholar 

  18. Mead RJ, Peterson MB, Welty JD: Sarcolemmal and sarcoplasmic reticular ATPase activities in the failing canine heart. Circ Res 29: 14–20, 1971

    Google Scholar 

  19. Gwathmey JK, Morgan JP: Altered calcium handling in experimental pressure overload hypertrophy in the ferret. Circ Res 57: 836–843, 1985

    Google Scholar 

  20. Ito Y, Suko J, Chidsey CA: Intracellular calcium and myocardial contractility. V. Calcium uptake of sarcoplasmic reticulum fractions in hypertrophied and failing rabbit hearts. J Molec Cell Cardiol 6: 237–247, 1974

    Google Scholar 

  21. Limas CJ, Cohn JN: Defective calcium transport by cardiac sarcoplasmic reticulum in spontaneously hypertensive rats. Circ Res 40: 1162–1169, 1977

    Google Scholar 

  22. Sordahl LA, McCollum WB, Wood WG, Schwartz A: Mitochondria and sarcoplasmic reticulum function in cardiac hypertrophy and failure. Am J Physiol 224: 497–502, 1973

    Google Scholar 

  23. Wei J-W, Janis RA, Daniel EE: Calcium accumulation and enzymatic activities of subcellular fractions from aortas and ventricles of genetically hypertensive rats. Circ Res 39: 133–140, 1976

    Google Scholar 

  24. Sulakhe PV, Dhalla NS: Excitation-contraction coupling in heart. VIII. Calcium accumulation in subcellular particles in congestive heart failure. J Clin Invest 50: 1019–1027, 1971

    Google Scholar 

  25. McCollum WB, Crow C, Harigaya S, Bajusz Scwartz A: Calcium binding by cardiac relaxing system isolated from myopathic Syrian hamsters (strains 14.6, 822.62 and 40.54). J Molec Cell Cardiol 1: 445–457, 1970

    Google Scholar 

  26. Lazarus MI, Colgan JA, Sachs HG: Quantitative light and electron microscopic comparison of the normal and cardiomyopathic Syrian hamster heart. J Molec Cell Cardiol 8: 431–441, 1976

    Google Scholar 

  27. Lucas-Heron B, Loirat MMJ, Ollivier B, Leoty C: Calcium-related defects in cardiac and skeletal muscles of dystrophic mice. Comp Biochem Physiol 86B: 295–301, 1987

    Google Scholar 

  28. Pierce GN, Kutryk MJB, Dhalla KS, Beamish RE, Dhalla NS: Biochemical alterations in heart after exhaustive swimming in rats. J Appl Physiol 57: 326–331, 1984

    Google Scholar 

  29. Conway G, Heazlitt RA, Fowler NO, Gabel M, Green S: The effect of hyperthyroidism on the sarcoplasmic reticulum and myosin ATPase of dog hearts. J Molec Cell Cardiol 8: 39–51, 1976

    Google Scholar 

  30. Ganguly PK, Pierce GN, Dhalla KS, Dhalla NS: Defective sarcoplasmic reticular calcium transport in diabetic cardiomyopathy. Am J Physiol E528–E535, 1983

  31. Penpargkul S, Fein F, Sonnenblick EH, Scheuer J: Depressed cardiac sarcoplasmic reticular function from diabetic rats. J Molec Cell Cardiol 13: 303–309, 1981

    Google Scholar 

  32. Dhalla NS, Pierce GN, Innes IR, Beamish RE: Pathogenesis of cardiac dysfunction in diabetes mellitus. Can J Cardiol 1: 263–281, 1985

    Google Scholar 

  33. Narayanan N: Effects of adrenalectomy and in vivo administration of dexamethasone on ATP-dependent calcium accumulation by sarcoplasmic reticulum from rat heart. J Molec Cell Cardiol 15: 7–15, 1983

    Google Scholar 

  34. Varley KG, Dhalla NS: Excitation-contraction coupling in heart. XII. Subcellular calcium transport in isoproterenol-induced myocardial necrosis. Exp Molec Path 19: 94–105, 1973

    Google Scholar 

  35. Panagia V, Pierce GN, Dhalla KS, Ganguly PK, Beamish RE, Dhalla NS: Adaptive changes in subcellular calcium transport during catecholamine-induced cardiomyopathy. J Molec Cell Cardiol 17: 411–420, 1985

    Google Scholar 

  36. Bing RJ, Tillmanns H, Fauvel J-M, Seeler K, Mao JC: Effect of prolonged alcohol administration on calcium transport in heart muscle of the dog. Circ Res 35: 33–38, 1974

    Google Scholar 

  37. Bellini O, Solcia E: Early and late sarcoplasmic reticulum changes in doxorubicin cardiomyopathy. An ultrastructural investigation with the zinc iodide-osmium tetroxide (ZIO) technique. Virchows Arch [Cell Path] 49: 137–152, 1985

    Google Scholar 

  38. Harris RN, Doroshow JH: Effect of doxorubicin-enhanced hydrogen peroxide and hydroxyl radical formation on calcium sequestration by cardiac sarcoplasmic reticulum. Biochem Biophys Res Commun 130: 739–745, 1985

    Google Scholar 

  39. McDonough KH: Calcium uptake by sarcoplasmic reticulum isolated from hearts of septic rats. Circ Shock 25: 291–297, 1988

    Google Scholar 

  40. Alto LE, Dhalla NS: Role of changes in microsomal calcium uptake in the effects of reperfusion of Ca2+-deprived rat hearts. Circ Res 48: 17–24, 1981

    Google Scholar 

  41. Lee SL, Dhalla NS: Subcellular calcium transport in failing hearts due to calcium deficiency and overload. Am J Physiol 231: 1159–1165, 1976

    Google Scholar 

  42. Rowe GT, Manson NH, Capland M, Hess ML: Hydrogen peroxide and hydroxyl radical mediation of activated leukocyte depression of cardiac sarcoplasmic reticulum. Participation of the cyclooxygenase pathway. Circ Res 53: 584–591, 1983

    Google Scholar 

  43. Korecky B, Ganguly PK, Elimban V, Dhalla NS: Muscle mechanics and Ca2+ transport in atrophic heart transplants in rat. Am J Physiol 251: H941-H950, 1986

    Google Scholar 

  44. Staley NA, Noren GR, Einzig S: Early alterations in the function of sarcoplasmic reticulum in a naturally occurring model of congestive cardiomyopathy. Cardiovasc Res 15: 276–281, 1981

    Google Scholar 

  45. Nagai R, Zarain-Herzberg A, Brandl CJ, Fujii J, Tada M, MacLennan DH, Alpert NR, Periasami M: Regulation of myocardial Ca2+-ATPase and phospholamban mRNA expression in response to pressure overload and thyroid hormone. Proc Natl Acad Sci 86: 2966–2970, 1989

    Google Scholar 

  46. Komura I, Kurabayashi M, Shibazaki Y, Takaku F, Yazaki Y: Molecular cloning and characterization of a Ca2+ + Mg2+-dependent adenosine triphosphatase from rat cardiac sarcoplasmic reticulum. Regulation of its expression by pressure overload and developmental stage. J Clin Invest 83: 1102–1108, 1989

    Google Scholar 

  47. de la Bastie D, Levitsky D, Rappaport L, Mercadier J-J, Marotte F, Wisnewsky C, Brovkovich V, Schwartz K, Lompre A-M: Function of the sarcoplasmic reticulum and expression of its Ca2+-ATPase gene in pressure overload-induced cardiac hypertrophy in the rat. Circ Res 66: 554–564, 1990

    PubMed  Google Scholar 

  48. Mercadier J-J, Lompre A-M, Due P, Boheler KR, Fraysse J-B, Wisnewsky C, Allen PD, Komadjda M, Schwartz K: Altered sarcoplasmic reticulum Ca2+-ATPase gene expression in the human ventricle during end-stage heart failure. J Clin Invest 85: 305–309, 1990

    Google Scholar 

  49. Feher JJ, Lebolt WR, Mason NH: Differential effect of global ischemia on the ryanodine-sensitive and ryanodine-insensitive calcium uptake of cardiac sarcoplasmic reticulum. Circ Res 65: 1400–1408

  50. O'Brien PJ: Microassay for malignant hyperthermia susceptibility: hypersensitive ligand-gating of the Ca-channel in muscle sarcoplasmic reticulum causes increased amounts and rates of Ca-release. Molec Cell Biochem 93: 53–59, 1990

    Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, Ontario Veterinary College, University of Guelph, N1G 2W1, Guelph, Ontario, Canada

    Peter James O'Brien, Hua Shen, Janice Weiler, Mehdi Mirsalimi & Richard Julian

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  1. Peter James O'Brien
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  4. Mehdi Mirsalimi
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O'Brien, P.J., Shen, H., Weiler, J. et al. Myocardial Ca-sequestration failure and compensatory increase in Ca-ATPase with congestive cardiomyopathy: kinetic characterization by a homogenate microassay using real-time ratiometric indo-1 spectrofluorometry. Mol Cell Biochem 102, 1–12 (1991). https://doi.org/10.1007/BF00232153

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