Summary
Transsarcolemmal water and ion movement during 1, 7.5, 15, and 30 min of total ischaemia was studied in suspensions of isolated rat ventricular myocytes, with a control ratio of about 1 of intracellular volume (ICV) to extracellular volume (ECV). In this preparation, contrary to the intact heart:
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1)
There is no external exchange of matter, 2) the sum of ICV and ECV remains constant and 3) ECV is homogeneous; no separate interstitial and intravascular compartments are present and no extracellular metabolite or ion gradients develop as may occur in the intact heart. We demonstrate that:
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1)
It is possible to make an ischaemic preparation of isolated myocytes with a procedure which causes only minimal mechanical damage to intact myocytes. The preparation allows measurement of ECV with the non-cardiac enzyme α-amylase as a macromolecular extracellular marker.
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2)
The time course of change of metabolites relevant to energy metabolism (creatinephosphate (CrP), creatine (Cr), ATP, ADP, inorganic phosphate Pi and lactate) is similar to that in the intact heart.
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3)
ECV has decreased and ICV increased by about 20% after 30 min of ischaemia.
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4)
Extracellular [Na+], [K+], [Cl−], and [Pi] increase, but not in proportion to the decrease of ECV. There is net efflux of K+, Pi, H+, and lactate−; efflux of K+ and Pi is quantitatively much less than influx of Na+ and Cl−.
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5)
Measured extracellular osmolality has increased with up to 70 mOsm/l after 30 min of ischaemia. The increase of extracellular [Iactate−], [Na+], [K+], [Cl−], [Pi] and the decrease of [glucose] account for the change of osmolality measured.
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6)
Summation of the electrical charges associated with measured increase of extracellular [Iactate−], [Na+], [K+], [Cl−], [Pi] shows a surplus of negative charge, which almost equals extracellular [Iactate−], suggesting an equally large increase of osmotically inactive H+ as the compensatory ion.
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7)
Blockade of anaerobic metabolism with iodoacetic acid (IAA) reduces efflux of Iactate and Pi but greatly amplifies influx of sodium and chloride and efflux of potassium.
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References
Armstrong SC, Ganote CE (1992) Effects of the protein phosphatase inhibitors okadaic acid and calyculin A on metabolically inhibited and ischaemic isolated myocytes. J Mol Cell Cardiol 24:869–884
Aksnes (1992) Why do ischemic and hypoxic myocardium lose potassium? J Mol Cell Cardiol 24:323–331
Cascio WE, Yan GX, Kleber AG (1992) Early changes in extracellular potassium in ischemic rabbit myocardium; The role of extracellular carbon dioxide accumulation and diffusion. Circ Res 70:409–422
Coronel R, Fiolet JWT, Wilms-Schopman FJG, Schaapherder AFM, Johnson TA, Gettes LS, Janse MJ (1988) Distribution of extracellular potassium and its relation to electrophysiologic changes during acute myocardial ischemia in the isolated perfused porcine heart. Circulation 77:1125–1135
Dresdner KP (1990) Ionic fluxes in ischemia and infarction. In: Rosen MR, Janse MJ, Wit AL (eds) Cardiac Electrophysiology: A Textbook. Futura Publishing Company Inc, Mount Kisco, NY, pp 695–718
van Echteld CJA, Kirkels JA, Eijgelshoven MHJ, van der Meer P, Ruigrok TJC (1991) Intracellular sodium during ischemia and calcium free perfusion: a23Na NMR study. J Mol Cell Cardiol 23:297–307
Fiolet JWT, Baartscheer A, Schumacher CA, Coronel R, ter Welle HF (1984) The change of the free energy of ATP hydrolysis during global ischemia and anoxia in the rat heart; its possible role in the regulation of transsarcolemmal sodium and potassium gradients. J Mol Cell Cardiol 16:1023–1036
Fiolet JWT, Baartscheer A, Schumacher CA, Krieger WJG (1985) Transmural inhomogeneity of energy metabolism during acute global ischemia in the isolated rat heart: dependence on environmental conditions. J Mol Cell Cardiol 17:87–92
Gassner RNA, Vaughan-Jones RD (1990) Mechanism of potassium efflux and action potential shortening during ischaemia in isolated mammalian cardiac muscle. J Physiol 431:713–741
Hill JL, Gettes LS (1980) Effect of acute coronary artery occlusion on local myocardial extracellular K+ activity in swine. Circulation 61:768–778
Hirche H, Franz C, Bos L, Bissig R, Lang R, Schram M (1980) Myocardial extracellular K+ and H+ increase and noradrenaline release as possible cause of early arrhythmias following acute coronary artery occlusion in pigs. J Mol Cell Cardiol 12:579–593
Kleber AG, Riegger CB, Janse MJ (1987) Extracellular K+ and H+ shifts in early ischemia: mechanisms and relation to changes in impulse propagation. J Mol Cell Cardiol 19 (Suppl V):35–44
Knopf H, Theising R, Moon CH, Hirche H (1990) Continuous determination of extracellular space and changes of K, Na, Ca, and H during global ischaemia in isolated rat hearts. J Mol Cell Cardiol 22:1259–1272
Kohmoto O, Krueger JA, Barry WH (1990) Activation of furosemide-sensitive K fluxes in myocytes by ouabain and recovery from metabolic inhibition. Am J Physiol 259:H962-H972
Krieger WJG, ter Welle HF, Fiolet JWT, Janse MJ (1984) Tissue osmolality, metabolic response, and reperfusion in myocardial ischemia. Basic Res Cardiol 79:562–571
Lazdunski M, Frelin C, Vigne P (1985) 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 17:1029–1042
Liu S, Jacob R, Piwnica-Worms D, Lieberman M (1987) (Na−K−2Cl) cotransport in cultured embrionic chick heart cells. Am J Physiol 253:C721-C730
McDonald TF, MacLeod DP (1973) Metabolism and the electrical activity of anoxic ventricular muscle. J Physiol 229:559–582
Mitani A, Shattock MJ (1992) Role of Na-activated K channel, Na−K−Cl cotransport, and Na−K pump in [K]e changes during ischemia in rat heart. Am J Physiol 263:H333-H340
Neubauer S, Balschi JA, Springer CS, Smith TW, Ingwall JS (1987) Intracellular Na+ accumulation in hypoxic vs. ischemic rat heart: evidence for Na+−H+ exchange. Circulation 76 (Suppl IV):56
Page E, Page E (1968) Distribution of ions and water between tissue compartments in the perfused left ventricle of the rat heart. Circ Res 21:435–446
Pike MM, Kitakaze M, Marban E (1990)23Na-NMR measurements of intracellular sodium in intact perfused ferret hearts during ischemia and reperfusion. Am J Physiol 259:H1767-H1773
Polimeni PI (1974) Extracellular space and ionic distribution in rat ventricle. Am J Physiol 227:676–683
Poole RC, Halestrap AP (1993) Transport of lactate and other monocarboxylates across mammalian plasma membranes. Am J Physiol 264:C761-C782
Schaapherder AFM, Schumacher CA, Coronel R, Fiolet JWT (1990) Transmural inhomogeneity of extracellular [K+] and pH and myocardial energy metabolism in the isolated rat heart during acute global ischemia; dependence on gaseous environment. Basic Res Cardiol 85:33–44
Tani M, Neely JR (1989) Role of intracellular Na+ in Ca2+ overload and depressed recovery of ventricular function of reperfused ischemic rat heart. Circ Res 65:1045–1056
Tranum-Jensen J, Janse MJ, Fiolet JWT, Krieger WJG, Naumann-d'Alnoncourt C, Durrer D (1981) Tissue osmolality, cell swelling, and reperfusion in acute regional myocardial ischemia in the isolated porcine heart. Circ Res 49:364–381
Tseng GN (1992) Cell swelling increases membrane conductance of canine cardiac cells: evidence for a volume-sensitive Cl channel. Am J Physiol 262:C1056-C1068
Van der Heide RS, Rim D, Hohl CM, Ganote CE (1990) An in vitro model of myocardial ischemia utilizing isolated adult rat myocytes. J Mol Cell Cardiol 22:165–181
Weiss J, Shine KI (1982) Extracellular K+ during myocardial ischemia in isolated rabbit heart. Am J Physiol 242:H619-H628
ter Welle HF, Baartscheer A, Fiolet JWT, Schumacher CA (1988) The cytoplasmic free energy of ATP hydrolysis in isolated rod-shaped rat ventricular myocytes. J Mol Cell Cardiol 20:435–441
Wiberg GS, Tuba J (1955) On rat serum amylase III. The contribution by various tissues to serum amylase activity. Canad J Biochem Physiol 33:817–825
Wilensky RL, Tranum-Jensen J, Coronel R, Wilde AAM, Fiolet JWT, Janse MJ (1986) The subendocardial border zone during acute ischemia of the rabbit heart: an electrophysiologic correlative study. Circulation 74:1137–1146
Wilde AAM, Kleber AG (1986) The combined effects of hypoxia, high K+, and acidosis on the intracellular sodium activity and resting potential in guinea pig papillary muscle. Circ Res 58:249–256
Wilde AAM, Escande D, Schumacher CA, Thuringer D, Mestre M, Fiolet JWT, Janse MJ (1990) Potassium accumulation in the globally ischemic mammalian heart; a role for the ATP-sensitive potassium channel. Circ Res 67:835–843
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Fiolet, J.W.T., Schumacher, C.A., Baartscheer, A. et al. Osmotic changes and transsarcolemmal ion transport during total ischaemia of isolated rat ventricular myocytes. Basic Res Cardiol 88, 396–410 (1993). https://doi.org/10.1007/BF00795407
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DOI: https://doi.org/10.1007/BF00795407