Volume regulation and plasma membrane injury in aerobic, anaerobic, and ischemic myocardium in vitro. Effects of osmotic cell swelling on plasma membrane integrity.

The relationship between cell swelling and plasma membrane disruption has been evaluated in thin myocardial slices incubated in oxygenated or anoxic Krebs-Ringer phosphate media. Electron microscopy and measurements of inulin-diffusible space were used to monitor plasma membrane integrity. Inulin is excluded from the intracellular space of intact cells; therefore, an increase in tissue inulin content is an excellent marker of loss of plasma membrane integrity. Cell volume was increased during exposure of aerobic slices to hypotonic media, but the inulin-diffusible space was not increased and electron micrographs showed no detectable plasma membrane alterations. Likewise, during prolonged anoxic isotonic incubation, no evidence of plasma membrane damage was observed. Incubation in anoxic hypotonic media for 60 minutes resulted in a larger increase in cell volume than under aerobic conditions, but plasma membrane integrity was maintained. Extended anoxic hypotonic incubation (300 minutes) produced no further change in tissue water, but the inulin-diffusible space was increased and electron micrographs revealed breaks in the plasma membranes primarily in association with large subsarcolemmal blebs. Likewise, myocardial slices incubated in isotonic anoxic media for 240 minutes and hypotonic anoxic media for 60 minutes had an increased inulin-diffusible space and the ultrastructural appearance was similar. This ultrastructural appearance is indistinguishable from that observed in myocytes lethally injured by ischemia. Measurements of tissue osmolarity during total ischemia showed that osmotically induced cell swelling could occur in ischemic myocardium prior to the onset of plasma membrane disruption. Our results indicate that cell swelling per se is incapable of rupturing plasma membranes; however, after prolonged periods of energy deficiency, the plasma membrane or its cytoskeletal scaffold become injured, which allows the membrane to rupture if the cell is swollen, as might occur during ischemia or reperfusion.

[1]  R. Jennings,et al.  Electrolytes of Damaged Myocardial Mitochondria 1 , 1970, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[2]  R. Jennings.  Ultrastructural changes in acute myocardial ischemia. , 1973, The Proceedings of the Institute of Medicine of Chicago.

[3]  R. Jennings,et al.  Structural Changes in Myocardium During Acute Ischemia , 1974, Circulation research.

[4]  M. Rabinowitz,et al.  Removal of Z-lines and alpha-actinin from isolated myofibrils by a calcium-activated neutral protease. , 1975, The Journal of biological chemistry.

[5]  W. Nayler,et al.  Irreversible myocardial injury in anoxic perfused rat hearts. , 1975, The American journal of pathology.

[6]  P. Polimeni,et al.  Expansion of Extracellular Space in the Nonischemic Zone of the Infarcted Heart and Concomitant Changes in Tissue Electrolyte Contents in the Rat , 1975, Circulation research.

[7]  R. Jennings,et al.  Experimental myocardial ischemic injury. II. Effect of in vivo ischemia on dog heart slice function in vitro. , 1976, Journal of molecular and cellular cardiology.

[8]  R. Jennings,et al.  Experimental myocardial ischemic injury. I. A comparison of Stadie-Riggs and free-hand slicing techniques on tissue ultrastructure, water and electrolytes during in vitro incubation. , 1976, Journal of molecular and cellular cardiology.

[9]  C. Halverson,et al.  Structural changes in the freeze-fractured sarcolemma of ischemic myocardium. , 1977, The American journal of pathology.

[10]  G. Bullock,et al.  The oxygen paradox and the calcium paradox: two facets of the same problem? , 1978, Journal of molecular and cellular cardiology.

[11]  J. Lowe,et al.  Relation between high energy phosphate and lethal injury in myocardial ischemia in the dog. , 1978, The American journal of pathology.

[12]  J. Farber,et al.  Ischemic myocardial cell injury. Prevention by chlorpromazine of an accelerated phospholipid degradation and associated membrane dysfunction. , 1979, The American journal of pathology.

[13]  P. Cala.  Volume regulation by Amphiuma red blood cells. The membrane potential and its implications regarding the nature of the ion-flux pathways , 1980, The Journal of general physiology.

[14]  P. A. Tornheim.  Use of a vapor pressure osmometer to measure brain osmolality , 1980, Journal of Neuroscience Methods.

[15]  J. L. Hill,et al.  Effect of Acute Coronary Artery Occlusion on Local Myocardial Extracellular K+ Activity in Swine , 1980, Circulation.

[16]  B. Geiger,et al.  Immunoelectron microscope studies of membrane-microfilament interactions: distributions of alpha-actinin, tropomyosin, and vinculin in intestinal epithelial brush border and chicken gizzard smooth muscle cells , 1981, The Journal of cell biology.

[17]  W. Dayton,et al.  Immunocytochemical localization of a calcium-activated protease in skeletal muscle cells. , 1981, Experimental cell research.

[18]  D. Durrer,et al.  Tissue Osmolality, Cell Swelling, and Reperfusion in Acute Regional Myocardial Ischemia in the Isolated Porcine Heart , 1981, Circulation research.

[19]  J. Willerson,et al.  Abnormalities of volume regulation and membrane integrity in myocardial tissue slices after early ischemic injury in the dog: effects of mannitol, polyethylene glycol, and propranolol. , 1981, The American journal of pathology.

[20]  A. Somlyo,et al.  Primary role of sarcoplasmic reticulum in phasic contractile activation of cardiac myocytes with shunted myolemma , 1981, The Journal of cell biology.

[21]  R. Jennings,et al.  Total Ischemia in Dog Hearts, in Vitro: 2. High Energy Phosphate Depletion and Associated Defects in Energy Metabolism, Cell Volume Regulation, and Sarcolemmal Integrity , 1981, Circulation research.

[22]  N. Shaikh.  Time Course of Changes in Porcine Myocardial Phospholipid Levels during Ischemia: A Reassessment of the Lysolipid Hypothesis , 1981, Circulation research.

[23]  R. Jennings,et al.  Total Ischemia in Dog Hearts, in Vitro: 1. Comparison of High Energy Phosphate Production, Utilization, and Depletion, and of Adenine Nucleotide Catabolism in Total Ischemia in Vitro vs. Severe Ischemia in Vivo , 1981, Circulation research.

[24]  K. Chien,et al.  Phospholipid Alterations in Canine Ischemic Myocardium: Temporal and Topographical Correlations with Tc‐99m‐PPi Accumulation and an in Vitro Sarcolemmal Ca2+ Permeability Defect , 1981, Circulation research.

[25]  R. Jennings,et al.  Lethal myocardial ischemic injury. , 1981, The American journal of pathology.

[26]  K I Shine,et al.  Extracellular K+ accumulation during myocardial ischemia in isolated rabbit heart. , 1982, The American journal of physiology.

[27]  M. Knepper.  Measurement of osmolality in kidney slices using vapor pressure osmometry. , 1982, Kidney international.

[28]  Farber Jl.  Biology of disease: membrane injury and calcium homeostasis in the pathogenesis of coagulative necrosis. , 1982, Laboratory investigation; a journal of technical methods and pathology.

[29]  P. Corr,et al.  Pathophysiological concentrations of lysophosphatides and the slow response. , 1982, The American journal of physiology.

[30]  P. Corr,et al.  Incorporation of Radiolabeled Lysophosphatidyl Choline into Canine Purkinje Fibers and Ventricular Muscle: Electrophysiological, Biochemical, and Autoradiographic Correlations , 1982, Circulation research.

[31]  J. Farber.  Biology of disease: membrane injury and calcium homeostasis in the pathogenesis of coagulative necrosis. , 1982, Laboratory investigation; a journal of technical methods and pathology.

[32]  S. Singer,et al.  Immunoelectron microscopic studies of desmin (skeletin) localization and intermediate filament organization in chicken cardiac muscle , 1983, The Journal of cell biology.

[33]  Price Mg,et al.  Intermediate filaments in striated muscle. A review of structural studies in embryonic and adult skeletal and cardiac muscle. , 1983 .

[34]  R. Jennings,et al.  Comparison of the effect of ischaemia and anoxia on the sarcolemma of the dog heart. , 1983, European heart journal.

[35]  C. Ganote.  Contraction band necrosis and irreversible myocardial injury. , 1983, Journal of molecular and cellular cardiology.

[36]  J. Siliciano,et al.  Vinculin is a component of an extensive network of myofibril-sarcolemma attachment regions in cardiac muscle fibers , 1983, The Journal of cell biology.

[37]  N. Sperelakis,et al.  The membrane systems and cytoskeletal elements of mammalian myocardial cells. , 1983, Cell and muscle motility.

[38]  A. Kléber,et al.  Resting Membrane Potential, Extracellular Potassium Activity, and Intracellular Sodium Activity during Acute Global Ischemia in Isolated Perfused Guinea Pig Hearts , 1983, Circulation research.

[39]  J. W. Sanger,et al.  Intermediate filaments in striated muscle. A review of structural studies in embryonic and adult skeletal and cardiac muscle. , 1983, Cell and muscle motility.

[40]  B. Geiger,et al.  The dynamics of cytoskeletal organization in areas of cell contact. , 1984, Cell and muscle motility.

[41]  R. Jennings,et al.  Relationship between lysophospholipid accumulation and plasma membrane injury during total in vitro ischemia in dog heart. , 1984, Journal of molecular and cellular cardiology.

[42]  K. Chien,et al.  Accumulation of Unesterified Arachidonic Acid in Ischemic Canine Myocardium: Relationship to a Phosphatidylcholine Deacylation‐Reacylation Cycle and the Depletion of Membrane Phospholipids , 1984, Circulation research.