TISSUE HYPOXIA AND ISCHEMIA ADVANCES IN EXPERIMENTAL MEDICINE AND BIOLOGY Editorial Board: Nathan Back State University of New York at Buffalo N. R. Di Luzio Tulane University School of Medicine Bernard Halpern College de France and Institute of Immuno·Biology Ephraim Katchalski The Weizmann Institute of Science David Kritchevsky Wistar Institute Abel Lajtha New York State Research Institute for Neurochemistry and Drug Addiction Rodolfo Paoletti University of Milan Recent Volumes in this Series Volume 73A THE RETICULOENDOTHELIAL SYSTEM IN HEALTH AND DISEASE: Functions and Characteristics Edited by Sherwood M. Reichard, Mario R. Escobar, and Herman Friedman. 1976 Volume 73B THE RETICULOENDOTHELIAL SYSTEM IN HEALTH AND DISEASE: Immunologic and Pathologic Aspects Edited by Herman Friedman, Mario R. Escobar, and Sherwood M. Reichard. 1976 Volume 74 IRON AND COPPER PROTEINS Edited by Kerry T. Yasunobu, Howard F. Mower, and Osamu Hayaishi .1976 Volume 75 OXYGEN TRANSPORT TO TISSUE-II Edited by Jiirgen Grote, Daniel Reneau, and Gerhard Thews. 1976 Volume 76A PURINE METABOLISM IN MAN II: Regulation of Pathways and Enzyme Defects Edited by Mathias M. Muller, Erich Kaiser, and J. Edwin Seegmiller. 1977 Volume 76B PURINE METABOLISM IN MAN II: Physiology, Pharmacology, and Clinical Aspects Edited by Mathias M. Muller, Erich Kaiser, and J. Edwin Seegmiller. 1977 Volume 77 RETINITIS PIGMENTOSA: Clinical Implications of Current Research Edited by Maurice B. Landers, III, Myron L. Wolbarsht, John E. Dowling, and Alan M. Laties • 1977 Volume 78 TISSUE HYPOXIA AND ISCHEMIA Edited by Martin Reivich, Ronald Coburn, Sukhamay Lahirl, and Britton Chance. 1977 Volume 79 ELASTIN AND ELASTIC TISSUE Edited by Lawrence B. Sandberg, William R. Gray, and Carl Franzblau • 1977 Volume 80 COMPARATIVE ENDOCRINOLOGY OF PROLACTIN Edited by Horst-Dieter Dellmann, J. Alan Johnson, and David M. Klachko • 1977 TISSUE HYPOXIA AND ISCHEMIA Edited by Martin Reivich, Ronald Coburn, Sukhamay Lahiri, and Britton Chance University of Pennsylvania, Philadelphia PLENUM PRESS • NEW YORK AND LONDON Library of Congress Cataloging in Publication Data Main entry under title: Tissue hypoxia and ischemia. (Advances in experimental medicine and biology; v. 78) Proceedings of a symposium held at the Annenherg Center of the University of Pennsylvania Aug. 13-14, 1976; sponsored by the Respiratory Physiology Group, Dept. of Physiology, School of Medicine, University of Pennsylvania, and others. Includes index. 1. Cerebral ischemia-Congresses. 2. Oxygen in the body-Congresses. 3. Chemo receptors-Congresses. 4. Vascular smooth muscle-Congresses. I. Reivich, Martin. II. Pennsylvania. University. Respiratory Physiology Group. IlL Series [DNLM: 1. Anoxia-Metabolism. 2. Ischemia, Transient-Metabolism. S991t v. 781976] RC388.5.T57 611 '.01 76-53008 ISBN-13: 978-1-4615-9037-8 e-ISBN-13: 978-1-4615-9035-4 001: 10.1007/978-1-4615-9035-4 Proceedings of a symposium on Tissue Hypoxia and Ischemia held at the Annenberg Center of the University of Pennsylvania, August 13-14,1976 © 1977 Plenum Press, New York Softcover reprint of the hardcover 1s t edition 1977 A Division of Plenum Publishing Corporation 227 West 17th Street, New York, N. Y. 10011 All rights reserved No part of this book may be reproduced, stored in a retrieval system, or transmitted, in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher Sponsors The Respiratory Physiology Group Department of Physiology School of Medicine University of Pennsylvania Cardiopulmonary Section Department of Medicine School of Medicine University of Pennsylvania Johnson Research Foundation University of Pennsylvania Cerebrovascular Research Center School of Medicine University of Pennsylvania Head Injury Center School of Medicine University of Pennsylvania Institute for Environmental Medicine School of Medicine University of Pennsylvania International Society of Oxygen Transport to Tissue This symposium was supported by Ayerst Laboratory Preface This monograph contains the proceedings of a symposium entitled, "Tissue Hypoxia and Ischemia," which was held at the Annenberg Center of the University of Pennsylvania on August 13 and 14, 1976. The symposium was jointly sponsored by the following groups at the University of Pennsylvania: the Respiratory Physiology Group of the Department of Physio logy, the Cardiopulmonary Section of the Department of Medicine, the Johnson Research Foundation, the Cerebrovascular Research Center of the Department of Neurology, the Head Injury Center of the Department of Neurosurgery, the. Institute for Environ mental Medicine, and the International Society on Oxygen Transport to Tissues. Its purpose was to promote an interdisciplinary discussion of oxygen sensors in various tissues and their mechanism of action as well as to examine the deleterious effects of hypoxia and ischemia with special reference to the brain. There were four sessions, one on the biochemistry of physi ologic oxygen sensors, two on the mechanism of oxygen sensing in tissues and one on the circulatory and metabolic aspects of cerebral hypoxia and ischemia. In the first session, conceptual problems concerning what constitutes a molecular oxygen sensor and the transduction pro cess were considered. In addition, the oxygen sensing character istics of microsomal enzymes were discussed as well as micro somal oxygenase reactions, in particular those in which cyto chrome P-450 plays a central role. The role of hydrogen peroxide formation in oxidation-reduction reactions involving the microsomes was explored. Other molecules which were con sidered as possible oxygen sensors were monoxygenases, myo globin and hemoglobin. The reactions and kinetics of these oxygenated hemeproteins were examined. There was also discussion of the peroxisomal enzymes; catalase and three oxidases (urate, L-a-hydroxyacid and D-aminoacid oxidases) with emphasis on their vii PREFACE properties which are important under physiologic conditions. Mitochondrial production of superoxide radicals and hydrogen peroxide, the oxygen dependence of this production and the physiologic relevance of these substances at the cellular level were considered. The second session dealt with the mechanism of oxygen sensing. Data concerning the bioelectric activity of chemo receptors and the effect of acetylcholine release on chemo receptor function was presented. Oxygen tension sensors of vascular smooth muscle were examined and a hypothesis to explain the production of oxygen dependent mechanical tension in vascular smooth muscle was put forth. Evidence was presented that the effect of hypoxia may be mediated by a mechanism other than inhibition of aerobic energy production. The mechanism of oxygen induced contraction of the ductus arteriosus and the roles of ATP, calcium ion and prostaglandins in this system were dis cussed. The sensing of oxygen tension in the pulmonary circula tion and the circulatory effects of tissue oxygen sensors, particularly in regard to coronary blood flow, were considered. The adenosine hypothesis for the regulation of blood flow in cardiac and skeletal muscle was critically examined. In the third session the examination of the mechanism of oxygen sensing in tissues was continued. The oxygen linked response of the carotid chemoreceptors and the interaction of hypoxic and hypercapnic stimuli were discussed. Data from micro electrode studies of the effects of changes in oxygen and carbon dioxide tension, temperature and osmolarity on carotid body cells were presented and the mechanism by which the chemo receptors sense changes in arterial oxygen and carbon dioxide tension were examined. The role of catecholamines and cyclic AMP in the chemoreception process of the carotid body was considered. The fourth session was concerned with the circulatory and metabolic aspects of cerebral hypoxia and ischemia. The character istic metabolic features of hypoxic hypoxia both at normal and reduced perfusion pressures as well as of incomplete and complete ischemia and how these metabolic changes relate to irreversible neuronal damage was discussed. Data demonstrating the presence of increased energy consumption and glucose metabolism in the brain following ischemia of transient duration was presented. Regional changes in energy metabolism and glycolysis in incom plete ischemia were also considered. The effects of ischemia of the cerebral cortex on other regions of the brain and spinal cord were examined in regard to cyclic nucleotide levels. The changes in tissue P02, ion fluxes and redox state produced by cerebral hypoxia and ischemia were discussed. Consideration was given to intracellular events possibly marking irreversible PREFACE injury following ischemia. The cerebral hemodynamic and metabolic alterations that occur in patients with cerebrovascular accidents and in animal models of strokes were examined. The effects of hypovolemic shock on cerebral blood flow and its regulation as well as on brain metabolism and mitochondrial function were dis cussed. The organ1z1ng committee would like to express its appreci ation to all those who participated in this symposium and helped make it a most successful one. We would also like to thank Ayerst Laboratory which generously provided support for this symposium. The Organizing Committee Contents THE BIOCHEMISTRY OF PHYSIOLOGIC OXYGEN SENSORS Chairman: B. Chance What is a molecular oxygen sensor? What is a transduction process? F.F. Jobsis.................................................... 3 The oxygen sensing characteristics of microsomal enzymes. R. W. Estabrook and J. Werringloer ........................ 19 0 • • • • Oxygen sensing heme proteins: monoxygenases, myoglobin and hemoglobin. I.C. Gunsalus, S.G. Sligar, T. Nordlund and H. Frauenfelder............................................... 37 Peroxisomal enzymes and oxygen metabolism in liver. H. Sies....................................................... Sl Mitochondrial production of superoxide radical and hydrogen peroxide. A. Boveris.................................................... 67 MECHANISM OF OXYGEN SENSING IN TISSUES Chairman: R.F. Coburn Chemoreception and transduction on neuronal models. N. Chalazoni tis. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8S Oxygen tension sensors in vascular smooth muscle. R.F. Coburn ..... 101 0 ••••••••••••••••••••••••••••••••••••••••• 0'" xi xii CONTENTS Comments on: oxygen tension sensors in vascular smooth muscle. R. J. Paul.................................................... 117 Mechanism of oxygen induced contraction of ductus arteriosus. F.S. Fay, P. Nair and W.J. Whalen ............................ 123 Prostaglandins and the control of muscle tone in the ductus arteriosus. F. Coceani, P.M. Olley, I. Bishai, E. Bodach, J. Heaton, M. Nashat and E. White............................ 135 The sensing of oxygen tension in the pulmonary circulation. A. P. Fishman................................................. 143 The sensing of oxygen tension in the pulmonary circulation. Discussion. N.C. Staub ................................................... 151 Circulatory effects of tissue oxygen tension sensors. R.M. Berne and R. Rubio ...................................... 163 Bioassay and pharmacologic evaluation of the adenosine hypothesis. F . J. Haddy................................................... 175 MECHANISM OF OXYGEN SENSING IN TISSUES Chairman: S. Lahiri Introductory remarks: oxygen linked response of carotid chemoreceptors. S. Lahiri .................................................... 185 Convergence of stimuli in arterial chemoreceptors. R.W. Torrance ................................................ 203 Intracellular studies of carotid body cells: effects of temperature, "natural" stimuli and chemical substances. C. Eyzaguirre, M. Baron and R. Gallego ....................... 209 Tissue P02 in the cat carotid body and related functions. W.J. Whalen and P. Nair ...................................... 225