Abstract
Idiopathic dilated cardiomyopathy is associated with derangement of myocardial sarcoplasmic Ca-homeostasis and energy production. The molecular mechanism for these changes is unknown. Accordingly, we used genetic and experimentally-induced models of canine dilated cardiomyopathy and tested the hypothesis that these metabolic changes resulted from altered gene expression, as indicated by mRNA content. We studied dilated cardiomyopathy occurring naturally (n=9) in Doberman pinschers, and in dogs subjected to rapid ventricular pacing (n=5), in comparison with normal dogs (n=9). We determined content and integrity of mRNA's using Northern and slot blotting, and measured activities of their translated product for the Ca-release channel and Ca-ATPase of sarcoplasmic reticulum, lactate dehydrogenase of glycolysis, citrate synthase of the tricarboxylic acid cycle, and for myoglobin, ATP-synthetase and the adenine nucleotide transporter, which are integral in oxidative phosphorylation. We found that, whereas both mRNA content and enzyme activity for markers of Ca-cycling, glycolysis, and oxidative phosphorylation were downregulated (20–80%) in dilated cardiomyopathy, they were upregulated (10–15%) for tricarboxylic acid cycling and for ribosomal RNA. RNA from cardiomyopathic tissue was up to 50% more degraded than for normal hearts in association with a 150% increase in ribonuclease activity. Downregulation of the Ca-cycle was asymmetric, with the Ca-channel being 65% more affected than the Ca-ATPase. This work supports the general paradigm that transcriptional and translational responses to pathophysiology are major determinants of the metabolic response seen in cardiac failure.
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Katz AM: Cardiomyopathy of overload. A major determinant of prognosis in congestive heart failure. New Eng J Med 322: 100–110, 1987
Gwathmey JKL, Copelas R, MacKinnon R, Shoer FJ, Feldman MD, Grossman W, Morgan JP: Abnormal intracellular calcium handling in myocardium from patients with end-stage heart failure. Circ Res 61: 70–76, 1987
Mercardier JJ, Lompre AM, Duc P, Boheler KR, Fraysse J-B, Wissnewsky C, Allen PD, Komajda M, Schwartz K: Altered sarcoplasmic reticulum Ca-ATPase gene expression in the human ventricle during end-stage heart failure. J Clin Invest 85: 305–309, 1990
Brillantes A-M, Allen P, Takahashi T, Izumo S, Marks AR: Differences in cardiac calcium release channel (ryanodine receptor) expression in myocardium from patients with end-stage heart failure caused by ischemic versus dilated cardiomyopathy. Circ Res 71: 18–26, 1992
Arai M, Alpert NR, MacLennan DH, Barton P, Periasamy M: Alterations in sarcoplasmic reticulum gene expression in human heart failure. A possible mechanism for alterations in systolic and diastolic properties of the failing myocardium. Circ Res 72: 463–469, 1993
Takeda N, Tanamura A, Iwai T, Nakamura I, Kato M, Ohkubo T, Noma K: Mitochondrial DNA deletion in human myocardium. Mol Cell Biochem 119: 105–108, 1993
Ozawa T, Sugiyama S, Tanaka M: Mitochondrial DNA mutations as the etiology of cardiomyopathy. In: M. Tada and Boca Raton (eds). Molecular Biology of the Myocardium. CRC Press, 1992, pp 137–152
Maurer I, Zierz S: Myocardial respiratory chain enzyme activities in idiopathic dilated cardiomyopathy, and comparison with those in atherosclerotic coronary artery disease and valvular aortic stenosis. Am J Cardiol 72: 428–433, 1993
Limas CJ, Olivari M-T, Goldenberg IF, Levine TB, Bendit DG, Simon A: Calcium uptake by cardiac sarcoplasmic reticulum in human dilated cardiomyopathy. Cardiovasc Res 21: 601–605, 1987
Movsesian MA, Bristow MR, Krall J: Ca2+ uptake by cardiac sarco-plasmic reticulum from patients with idiopathic dilated cardiomyopathy. Circ Res 65: 1141–1144, 1989
O'Brien PJ, O'Grady M, McCutcheon LJ, Shen H, Nowack LN, Horne RD, Mirsalimi SM, Julian RJ, Grima EA, Moe GW, Armstrong PW: Myocardial myoglobin deficiency in various animal models of congestive heart failure. J Mol Cell Cardiol 24: 721–730, 1992
McCutcheon LJ, Cory CR, Nowack L, Shen H, Mirsalimi M, Lahucky R, Kovac L, O'Grady M, Horne MR, O'Brien PJ: Respiratory chain defect of myocardial mitochondria in idiopathic cardiomyopathy of Doberman pinscher dogs. Can J Physiol Pharmacol 70: 1529–1533, 1992
Cory CR, McCutcheon LJ, O'Grady M, Pang AW, Geiger JD, O'Brien PJ: Compensatory downregulation of myocardial Ca channel in SR from dogs with heart failure. Am J Physiol 264: H926–937, 1993
Cory CR, Shen H, O'Brien PJ: Compensatory asymmetry in down-regulation and inhibition of the Ca-cycle in congestive heart failure produced in dogs by idiopathic dilated cardiomyopathy. J Molec Cell Cardiol 26: 173–184, 1994
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
O'Brien PJ, Shen H, Weiler J, Ianuzzo CD, Wittnich C, Moe GW, Armstrong PW: Cardiac and muscle fatigue owing to relative functional overload induced by excessive stimulation, hypersensitive excitation-contraction coupling, or diminished performance capacity correlates with sarcoplasmic reticulum failure. Can J Physiol Pharmacol 69: 262–268, 1991
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. Mol Cell Biochem 94: 113–119, 1990
Shen H, Mirsalimi SM, Weller JE, Julian RJ, O'Brien PJ: Effects of mild cardiac hypetrophy, induced by volume overload in turkeys on myocardial sarcoplasmic reticulum calcium-pump and calcium-channel activities and on the creatine kinase system. Am J Vet Res 52: 1527–1530, 1991
Bermudez VM, Miller RB, Rosendal S, Fernando MA, Johnson SH, O'Brien PJ: Measurement of the cytotoxic effects of different strains ofMycoplasma equigenitalium on the equine uterine tube using a calmodulin assay. Can J Vet Res 56: 331–338, 1992
Kamm RC, Smith AG, Lyons H: A fluorometric method for assay of RNase activity. Analytical biochemistry 37: 333–336, 1970
Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanadinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987
Sambrook J, Fritsch EF, Maniatis T: Molecular Clonging. A Laboratory Manual (2nd ed). Plainview: Cold Spring Harbor Lab Press, 1989
Neckelmann N, Li K, Wade RP, Shuster R, Wallace DC: cDNA sequence of a human skeletal muscle ADP/ATP translocator cDNA, and coevolution with mitochondrial DNA genes. Proc Natl Acad Sci USA 84: 7580–7584, 1987
Wallace DC, Ye J, Neckelmann SN, Singh G, Webster KA, Greenberg BD: Sequence analysis of cDNA's for the human and bovine ATP synthase β subunit: mitochondrial DNA genes sustain seventeen times more mutations. Curr Genet 12: 81–90, 1987
Annex BH, Kraus WE, Dohm GL, Williams RS: Am J Physiol 260: C266-C270, 1991
Chang E, Kunes J, Hamet P, Tremblay J: Increase of calmodulin activator in hypertension. Modulation by dietary sodium and calcium. Am J Hypertens 3: 210S-215S, 1990
Wenz I, Reissmann R, Bornig H, Hoppe H, Cumme G: Ultramicro-ELISA for calmodulin and anticalmodulin. Biomed Biochim Acta 1: 125–133, 1987
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O'Brien, P.J., Duke, A.L., Shen, H. et al. Myocardial mRNA content and stability, and enzyme activities of Ca-cycling and aerobic metabolism in canine dilated cardiomyopathies. Mol Cell Biochem 142, 139–150 (1995). https://doi.org/10.1007/BF00928935
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DOI: https://doi.org/10.1007/BF00928935