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. 1989 Mar;410:297–323. doi: 10.1113/jphysiol.1989.sp017534

The consequences of simulated ischaemia on intracellular Ca2+ and tension in isolated ferret ventricular muscle.

D G Allen 1, J A Lee 1, G L Smith 1
PMCID: PMC1190480  PMID: 2795481

Abstract

1. In order to study cellular events occurring in ischaemia, we have developed a method for simulating ischaemia in an isolated papillary muscle. Muscles were suspended in a chamber and changed from conventional superfusion with Tyrode solution to gas perfusion with 95% N2/5% CO2 (N2 gas perfusion), thus simultaneously stopping oxygenation and flow. Surface cells of the preparation were injected with the photoprotein aequorin in order to monitor intracellular free calcium concentration [( Ca2+]i). 2. Gas perfusion with 95% O2/5% CO2 (O2 gas perfusion) had little effect on the tension or Ca2+ transients. Superfusion with Tyrode solution equilibrated with 95% N2/5% CO2 (N2 Tyrode) caused tension to decline to 30-40% of control, but had little effect on the amplitude of the Ca2+ transients. N2 gas perfusion caused tension to fall more rapidly and to a lower level than superfusion with N2 Tyrode. Ca2+ transients showed a small initial decline followed by a slowly developing increase in magnitude and duration. 3. Long exposures to N2 gas perfusion caused tension to decline to very low levels and Ca2+ transients to increase to a maximum. After a variable length of time, resting tension began to increase. At approximately the same time, Ca2+ transients began to decrease and eventually disappeared. Resting Ca2+ increased during N2 gas perfusion and remained elevated when the Ca2+ transients had declined. These changes could be reversed by restarting superfusion with standard Tyrode or by perfusion with O2 gas. 4. N2 gas perfusion caused a depolarization of the resting potential and an abbreviation of the action potential. In a long exposure the action potential eventually failed. These changes could be reversed by restarting superfusion with standard Tyrode or by perfusion with O2 gas. 5. Many of the effects of N2 gas perfusion could be mimicked by the addition of 20 mM-lactic acid to the superfusing solution, which caused a profound reduction of tension and also an increase in the amplitude and duration of the Ca2+ transients. Calculation of the changes in intracellular pH caused by the addition of lactic acid suggest that the fall in intracellular pH produced by lactic acid was similar to that occurring in ischaemia. 6. Repeated exposures to N2 gas perfusion caused tension to fall more rapidly and an increased resting tension to develop more rapidly. The slowly developing rise in Ca2+ transients was abolished and a rise in resting Ca2+ occurred more quickly. 7. When muscles were quiescent, exposure to N2 gas perfusion caused an increase in resting light.(ABSTRACT TRUNCATED AT 400 WORDS)

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Selected References

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  1. Allen D. G., Blinks J. R., Prendergast F. G. Aequorin luminescence: relation of light emission to calcium concentration--a calcium-independent component. Science. 1977 Mar 11;195(4282):996–998. doi: 10.1126/science.841325. [DOI] [PubMed] [Google Scholar]
  2. Allen D. G., Morris P. G., Orchard C. H., Pirolo J. S. A nuclear magnetic resonance study of metabolism in the ferret heart during hypoxia and inhibition of glycolysis. J Physiol. 1985 Apr;361:185–204. doi: 10.1113/jphysiol.1985.sp015640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Allen D. G., Orchard C. H. Intracellular calcium concentration during hypoxia and metabolic inhibition in mammalian ventricular muscle. J Physiol. 1983 Jun;339:107–122. doi: 10.1113/jphysiol.1983.sp014706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Allen D. G., Orchard C. H. Myocardial contractile function during ischemia and hypoxia. Circ Res. 1987 Feb;60(2):153–168. doi: 10.1161/01.res.60.2.153. [DOI] [PubMed] [Google Scholar]
  5. Allen D. G., Orchard C. H. The effects of changes of pH on intracellular calcium transients in mammalian cardiac muscle. J Physiol. 1983 Feb;335:555–567. doi: 10.1113/jphysiol.1983.sp014550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bailey I. A., Williams S. R., Radda G. K., Gadian D. G. Activity of phosphorylase in total global ischaemia in the rat heart. A phosphorus-31 nuclear-magnetic-resonance study. Biochem J. 1981 Apr 15;196(1):171–178. doi: 10.1042/bj1960171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Barry W. H., Peeters G. A., Rasmussen C. A., Jr, Cunningham M. J. Role of changes in [Ca2+]i in energy deprivation contracture. Circ Res. 1987 Nov;61(5):726–734. doi: 10.1161/01.res.61.5.726. [DOI] [PubMed] [Google Scholar]
  8. Bers D. M., Ellis D. Intracellular calcium and sodium activity in sheep heart Purkinje fibres. Effect of changes of external sodium and intracellular pH. Pflugers Arch. 1982 Apr;393(2):171–178. doi: 10.1007/BF00582941. [DOI] [PubMed] [Google Scholar]
  9. Blanchard E. M., Solaro R. J. Inhibition of the activation and troponin calcium binding of dog cardiac myofibrils by acidic pH. Circ Res. 1984 Sep;55(3):382–391. doi: 10.1161/01.res.55.3.382. [DOI] [PubMed] [Google Scholar]
  10. Blinks J. R. Field stimulation as a means of effecting the graded release of autonomic transmitters in isolated heart muscle. J Pharmacol Exp Ther. 1966 Feb;151(2):221–235. [PubMed] [Google Scholar]
  11. Case R. B., Felix A., Castellana F. S. Rate of rise of myocardial PCO2 during early myocardial ischemia in the dog. Circ Res. 1979 Sep;45(3):324–330. doi: 10.1161/01.res.45.3.324. [DOI] [PubMed] [Google Scholar]
  12. Clusin W. T. Mechanism by which metabolic inhibitors depolarize cultured cardiac cells. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3865–3869. doi: 10.1073/pnas.80.12.3865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Cobbold P. H., Bourne P. K. Aequorin measurements of free calcium in single heart cells. 1984 Nov 29-Dec 5Nature. 312(5993):444–446. doi: 10.1038/312444a0. [DOI] [PubMed] [Google Scholar]
  14. Colquhoun D., Neher E., Reuter H., Stevens C. F. Inward current channels activated by intracellular Ca in cultured cardiac cells. Nature. 1981 Dec 24;294(5843):752–754. doi: 10.1038/294752a0. [DOI] [PubMed] [Google Scholar]
  15. Dahl G., Isenberg G. Decoupling of heart muscle cells: correlation with increased cytoplasmic calcium activity and with changes of nexus ultrastructure. J Membr Biol. 1980 Mar 31;53(1):63–75. doi: 10.1007/BF01871173. [DOI] [PubMed] [Google Scholar]
  16. De Mello W. C. Effect of intracellular injection of calcium and strontium on cell communication in heart. J Physiol. 1975 Sep;250(2):231–245. doi: 10.1113/jphysiol.1975.sp011051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dilly S. G., Lab M. J. Electrophysiological alternans and restitution during acute regional ischaemia in myocardium of anaesthetized pig. J Physiol. 1988 Aug;402:315–333. doi: 10.1113/jphysiol.1988.sp017206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Downar E., Janse M. J., Durrer D. The effect of acute coronary artery occlusion on subepicardial transmembrane potentials in the intact porcine heart. Circulation. 1977 Aug;56(2):217–224. doi: 10.1161/01.cir.56.2.217. [DOI] [PubMed] [Google Scholar]
  19. Ellis D., Thomas R. C. Direct measurement of the intracellular pH of mammalian cardiac muscle. J Physiol. 1976 Nov;262(3):755–771. doi: 10.1113/jphysiol.1976.sp011619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fabiato A., Fabiato F. Effects of pH on the myofilaments and the sarcoplasmic reticulum of skinned cells from cardiace and skeletal muscles. J Physiol. 1978 Mar;276:233–255. doi: 10.1113/jphysiol.1978.sp012231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gabel L. P., Bihler I., Dresel P. E. Contractility, metabolism and pharmacological reactions of isolated gas-perfused cat hearts. Circ Res. 1966 Nov;19(5):891–902. doi: 10.1161/01.res.19.5.891. [DOI] [PubMed] [Google Scholar]
  22. Gettes L. S., Reuter H. Slow recovery from inactivation of inward currents in mammalian myocardial fibres. J Physiol. 1974 Aug;240(3):703–724. doi: 10.1113/jphysiol.1974.sp010630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hess P., Metzger P., Weingart R. Free magnesium in sheep, ferret and frog striated muscle at rest measured with ion-selective micro-electrodes. J Physiol. 1982 Dec;333:173–188. doi: 10.1113/jphysiol.1982.sp014447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Ichihara K., Haga N., Abiko Y. Is ischemia-induced pH decrease of dog myocardium respiratory or metabolic acidosis? Am J Physiol. 1984 May;246(5 Pt 2):H652–H657. doi: 10.1152/ajpheart.1984.246.5.H652. [DOI] [PubMed] [Google Scholar]
  25. Janse M. J., Kléber A. G. Electrophysiological changes and ventricular arrhythmias in the early phase of regional myocardial ischemia. Circ Res. 1981 Nov;49(5):1069–1081. doi: 10.1161/01.res.49.5.1069. [DOI] [PubMed] [Google Scholar]
  26. Kass R. S., Lederer W. J., Tsien R. W., Weingart R. Role of calcium ions in transient inward currents and aftercontractions induced by strophanthidin in cardiac Purkinje fibres. J Physiol. 1978 Aug;281:187–208. doi: 10.1113/jphysiol.1978.sp012416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kentish J. C. The effects of inorganic phosphate and creatine phosphate on force production in skinned muscles from rat ventricle. J Physiol. 1986 Jan;370:585–604. doi: 10.1113/jphysiol.1986.sp015952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kimura J., Miyamae S., Noma A. Identification of sodium-calcium exchange current in single ventricular cells of guinea-pig. J Physiol. 1987 Mar;384:199–222. doi: 10.1113/jphysiol.1987.sp016450. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kléber A. G., Riegger C. B., Janse M. J. Electrical uncoupling and increase of extracellular resistance after induction of ischemia in isolated, arterially perfused rabbit papillary muscle. Circ Res. 1987 Aug;61(2):271–279. doi: 10.1161/01.res.61.2.271. [DOI] [PubMed] [Google Scholar]
  30. Lazdunski M., Frelin C., Vigne P. The sodium/hydrogen exchange system in cardiac cells: its biochemical and pharmacological properties and its role in regulating internal concentrations of sodium and internal pH. J Mol Cell Cardiol. 1985 Nov;17(11):1029–1042. doi: 10.1016/s0022-2828(85)80119-x. [DOI] [PubMed] [Google Scholar]
  31. Lee H. C., Smith N., Mohabir R., Clusin W. T. Cytosolic calcium transients from the beating mammalian heart. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7793–7797. doi: 10.1073/pnas.84.21.7793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Lee J. A., Allen D. G. The effects of repeated exposure to anoxia on intracellular calcium, glycogen and lactate in isolated ferret heart muscle. Pflugers Arch. 1988 Nov;413(1):83–89. doi: 10.1007/BF00581232. [DOI] [PubMed] [Google Scholar]
  33. MacKinnon R., Gwathmey J. K., Morgan J. P. Differential effects of reoxygenation on intracellular calcium and isometric tension. Pflugers Arch. 1987 Aug;409(4-5):448–453. doi: 10.1007/BF00583800. [DOI] [PubMed] [Google Scholar]
  34. Marban E., Kitakaze M., Kusuoka H., Porterfield J. K., Yue D. T., Chacko V. P. Intracellular free calcium concentration measured with 19F NMR spectroscopy in intact ferret hearts. Proc Natl Acad Sci U S A. 1987 Aug;84(16):6005–6009. doi: 10.1073/pnas.84.16.6005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Meech R. W., Thomas R. C. The effect of calcium injection on the intracellular sodium and pH of snail neurones. J Physiol. 1977 Mar;265(3):867–879. doi: 10.1113/jphysiol.1977.sp011749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nayler W. G., Poole-Wilson P. A., Williams A. Hypoxia and calcium. J Mol Cell Cardiol. 1979 Jul;11(7):683–706. doi: 10.1016/0022-2828(79)90381-x. [DOI] [PubMed] [Google Scholar]
  37. Neely J. R., Grotyohann L. W. Role of glycolytic products in damage to ischemic myocardium. Dissociation of adenosine triphosphate levels and recovery of function of reperfused ischemic hearts. Circ Res. 1984 Dec;55(6):816–824. doi: 10.1161/01.res.55.6.816. [DOI] [PubMed] [Google Scholar]
  38. Noma A. ATP-regulated K+ channels in cardiac muscle. Nature. 1983 Sep 8;305(5930):147–148. doi: 10.1038/305147a0. [DOI] [PubMed] [Google Scholar]
  39. Orchard C. H., Eisner D. A., Allen D. G. Oscillations of intracellular Ca2+ in mammalian cardiac muscle. Nature. 1983 Aug 25;304(5928):735–738. doi: 10.1038/304735a0. [DOI] [PubMed] [Google Scholar]
  40. Orchard C. H. The role of the sarcoplasmic reticulum in the response of ferret and rat heart muscle to acidosis. J Physiol. 1987 Mar;384:431–449. doi: 10.1113/jphysiol.1987.sp016462. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Reber W. R., Weingart R. Ungulate cardiac purkinje fibres: the influence of intracellular pH on the electrical cell-to-cell coupling. J Physiol. 1982 Jul;328:87–104. doi: 10.1113/jphysiol.1982.sp014254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Shen A. C., Jennings R. B. Myocardial calcium and magnesium in acute ischemic injury. Am J Pathol. 1972 Jun;67(3):417–440. [PMC free article] [PubMed] [Google Scholar]
  43. Smith G. L., Allen D. G. Effects of metabolic blockade on intracellular calcium concentration in isolated ferret ventricular muscle. Circ Res. 1988 Jun;62(6):1223–1236. doi: 10.1161/01.res.62.6.1223. [DOI] [PubMed] [Google Scholar]
  44. Steenbergen C., Murphy E., Levy L., London R. E. Elevation in cytosolic free calcium concentration early in myocardial ischemia in perfused rat heart. Circ Res. 1987 May;60(5):700–707. doi: 10.1161/01.res.60.5.700. [DOI] [PubMed] [Google Scholar]
  45. Szatkowski M. S., Thomas R. C. New method for calculating pHi from accurately measured changes in pHi induced by a weak acid and base. Pflugers Arch. 1986 Jul;407(1):59–63. doi: 10.1007/BF00580721. [DOI] [PubMed] [Google Scholar]
  46. Thandroyen F. T., McCarthy J., Burton K. P., Opie L. H. Ryanodine and caffeine prevent ventricular arrhythmias during acute myocardial ischemia and reperfusion in rat heart. Circ Res. 1988 Feb;62(2):306–314. doi: 10.1161/01.res.62.2.306. [DOI] [PubMed] [Google Scholar]
  47. Vaughan-Jones R. D., Lederer W. J., Eisner D. A. Ca2+ ions can affect intracellular pH in mammalian cardiac muscle. Nature. 1983 Feb 10;301(5900):522–524. doi: 10.1038/301522a0. [DOI] [PubMed] [Google Scholar]
  48. Wier W. G., Kort A. A., Stern M. D., Lakatta E. G., Marban E. Cellular calcium fluctuations in mammalian heart: direct evidence from noise analysis of aequorin signals in Purkinje fibers. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7367–7371. doi: 10.1073/pnas.80.23.7367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. de Hemptinne A., Marrannes R., Vanheel B. Influence of organic acids on intracellular pH. Am J Physiol. 1983 Sep;245(3):C178–C183. doi: 10.1152/ajpcell.1983.245.3.C178. [DOI] [PubMed] [Google Scholar]

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