Handbook of Experimental Pharmacology Volume 83

Editorial Board G.y' R. Born, P. Cuatrecasas, Research Triangle Park, NC H. Herken, A. Schwartz, Cincinnati, OH Calciumin Drug Actions

Contributors D. M. Bers, P. 1. R. Bevis, M. P. Blaustein, R. D. Bukoski, B. Ceccarelli, R. A. Chapman, S. Cockcroft, M. Crompton, A.W. Cuthbert, S. Ebashi, C. H. Evans, H. Fleisch, M. Fosset, M. L. Garcia, T. Godfraind, B. D. Gomperts, T. R. Hinds, P. Honerjager, M. Hugues, G. 1. Kaczorowski, U. Kikkawa, V. F. King, M. Lazdunski, B.A. Levine, I. MacIntyre, K.T. MacLeod, D. A. McCarron, 1. Meldolesi, C. Milet, C. Mourre, H. Nagamoto, T. Narahashi, Y. Nishizuka, Y Ogawa, T. Pozzan, 1. F. Renaud, G. Romey, H. Schmid-Antomarchi, M. Schramm, I. Schulz, T. 1. B. Simons, R. S. Slaughter, R. Towart, D. 1. Triggle, 1. Tunstall, F. F. Vincenzi, H. 1. Vogel, R.1. P. Williams, M. Zaidi

Editor P.R Baker

Springer-Verlag Berlin Heidelberg New York London Paris Tokyo PETER F. BAKER, Professor Sc. D., F.R.S. (t) Department of Physiology King's College, University of London, Strand, London WC2R 2LS, Great Britain

With 123 Figures

ISBN-13: 978-3-642-71808-3 e-ISBN-13: 978-3-642-71806-9 DOl: 10.1007/978-3-642-71806-9

Library of Congress Cataloging-in-Publication Data. Calcium in drug actions/contributors. D. M. Bers ... let al.]: editor, P. F. Baker. p. cm. -(Handbook of experimental pharmacology: v. 83) Includes bibliographies and index. 1. Calcium-Therapeutic use-Testing. 2. Calcium channels-Effect of drugs on. 3. Calcium-Receptors-Effect of drugs on. 4. Drugs-Physiological effect. I. Bers, D. M. II. Baker, Peter F. (Peter Frederick), 1939- . III. Series. [DNLM: 1. Calcium-physiology. WlHA51L v. 83/ QV 276 C1433] QP905.H3 vol. 83 [RM666.C6] 615' .1 s-cc19 [615'.2393] DNLM/DLC for Library of Congress

This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, re-use of illustrations, recitation, broadcasting, reproduction on microfilms or in other ways, and storage in data banks. Duplication of this publication or parts thereof is only permitted under the provisions of the German Copyright Law of September 9, 1965, in its version of June 24, 1985, and a copyright fee must always be paid. Violations fall under the prosecution act of the German Copyright Law. © Springer-Verlag Berlin Heidelberg 1988 Softcover reprint of the hardcover I st edition 1988 The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Product liability: The publisher can give no guarantee for information about drug dosage and application thereof contained in this book. In every individual case the respective user must check its accuracy by consulting other pharmaceutical literature.

2122/3130-543210 List of Contributors

D. M. BERS, Division of Biomedical Science, University of California, Riverside, CA 92521, USA P. J. R. BEVIS, Endocrine Unit, Department of Chemical Pathology, Royal Post• graduate Medical School, Hammersmith Hospital, Ducane Road, London W12 OHS, Great Britain M. P. BLAUSTEIN, University of Maryland School of Medicine, Department of Physiology, 655 West Baltimore Street, Baltimore, MD 21201, USA R. D. BUKOSKI, Division of Nephrology, Oregon Health Sciences University, 3181 SW Sam Jackson Park Rd., Portland, OR 97201, USA B. CECCARELLI, Center for the Study of Peripheral Neuropathies and Neuro• muscular Diseases, Department of Pharmacology, University of Milan, Via Vanvitelli, 32, 1-20129 Milan R. A. CHAPMAN, Department of Physiology, School of Veterinary Science, University of Bristol, Park Row, Bristol BS1 5LS, Great Britain S. COCKCROFT, Department of Experimental Pathology, University College London, University Street, London WC1E 6JJ, Great Britain M. CROMPTON, Department of Biochemistry, University College London, Gower Street, London WC1E 6BT, Great Britain A. W. CUTHBERT, Department of Pharmacology, University of , Hills Road, Cambridge CB22QD, Great Britain S. EBASHI, National Institute for Physiological Sciences, Myodaiji, Okazaki 444, Japan C. H. EVANS, Ferguson Laboratory for Orthopedic Research, University of Pittsburgh, School of Medicine, 986 Scaife Hall, Pittsburgh, PA 15261, USA H. FLEISCH, Department of Pathophysiology, University of Berne, Murten• strasse 35, CH-3010 Berne M. FOSSET, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F-06034 Nice Cedex M. L. GARCIA, Department of Membrane Biochemistry and Biophysics, Merck Institute for Therapeutic Research, P.O. Box 2000, Rahway, NJ 07065, USA VI List of Contributors

T. GoDFRAIND, Laboratoire de Pharmacodynamie Generale et de Pharmacologie, Universite Catholique de Louvain, UCL 7350, Ave. Emmanuel Mounier 73, B-1200 Brussels B. D. GoMPERTS, Department of Pathology, University College London, Gower Street, London WC1E 6BT, Great Britain T. R. HINDS, Department of Pharmacology, SJ-30, School of Medicine, University of Washington, Seattle, WA 98195, USA

P. HONERJAGER, Institut fUr Pharmakologie und Toxikologie der Technischen Universitat Munchen, Biedersteiner Strasse 29, D-8000 Munchen 40

M. HUGUES, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F -06034 Nice Cedex

G. J. KACZOROWSKI, Department of Membrane Biochemistry and Biophysics, Room 80N-32C, Merck Institute for Therapeutic Research, P.O. Box 2000, Rahway, NJ 07065, USA .

U. KIKKAWA, Department of Biochemistry, Kobe University School of Medicine, Chuo-ku, Kobe 650, Japan V. F. KING, Department of Membrane Biochemistry and Biophysics, Merck Institute for Therapeutic Research, P.O. Box 2000, Rahway, NJ 07065, USA

M. LAZDUNSKI, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F-06034 Nice Cedex

B. A. LEVINE, University of Oxford, Inorganic Chemistry Laboratory, South Parks Road, Oxford OX1 3QR, Great Britain I. MAcINTYRE, Endocrine Unit, Department of Chemical Pathology, Royal Postgraduate Medical School, Hammersmith Hospital, Ducane Road, London W12 OHS, Great Britain K. T. MACLEOD, Division of Biomedical Science, University of California, Riverside, CA 92521, USA D. A. MCCARRON, Division of Nephrology and Hypertension, Oregon Health Sciences University, 3181 SW Sam Jackson Park Road, Portland, OR 97201, USA J. MELDOLESI, University of Milan, Department of Pharmacology, Scientific Institute S. Raffaele, Via Olgettina, 60, 1-20132 Milan

C. MILET, Laboratoire de Physiologie Generale et Comparee, Centre National de la Recherche Scientifique, F -7 5231 Paris Cedex 5

C. MOURRE, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F-06034 Nice Cedex List of Contributors VII

H. NAGAMOTO, Tokushima Research Institute, Otsuka Pharmaceutical Co. Ltd., Tokushima, 771-01, Japan T. NARAHASHI, The Medical and Dental Schools, Department of Pharmacology, Northwestern University, 303 East Chicago Ave., Chicago, IL 60611, USA Y. NISHIZUKA, Department of Biochemistry, Kobe University School of Medicine, Chuo-ku, Kobe 650, Japan Y. OGAWA, Department of Pharmacology, Juntendo University School of Medicine, Hongo, Bunkyo-ku, Tokyo 113, Japan T. POZZAN, Department of General Pathology and CNR Center Membrane Physiology, University of Padova, Via Loredan, 16, I-Padova J. F. RENAUD, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F-06034 Nice Cedex G. ROMEY, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F-06034 Nice Cedex

H. SCHMID-ANTOMARCHI, Centre de Biochimie, Universite de Nice, Centre National de la Recherche Scientifique, Parc Valrose, 28, Ave. Valrose, F-06034 Nice Cedex M. SCHRAMM, Bayer AG, Institut fUr Pharmakologie, Aprather Weg 18a, Post• fach 101709, D-5600 Wuppertal I. SCHULZ, Max-Planck Institut fUr Biophysik, Kennedyallee 70, D-6000 Frank• furt/Main 70 T. J. B. SIMONS, Department of Physiology, King's College, University of London, Strand, London WC2R 2LS, Great Britain R. S. SLAUGHTER, Department of Membrane Biochemistry and Biophysics, Merck Institute for Therapeutic Research, P.O. Box 2000, Rahway, NJ 07065, USA R. TOWART, Miles Laboratories Ltd., Stoke Court, Stoke Poges, Bucks. SL24LY, Great Britain D. J. TRIGGLE, Department of Biochemical Pharmacology, School of Pharmacy, Faculty of Health Sciences, State University of New York at Buffalo, 313 Hochstetter Hall, Buffalo, NY 14260, USA J. TUNSTALL, Department of Physiology, University of Leicester, Leicester, Great Britain F. F. VINCENZI, Department of Pharmacology, SJ-30, School of Medicine, University of Washington, Seattle, WA 98195, USA H. J. VOGEL, University of Calgary, Division of Biochemistry, Department of Biological Sciences, 2500 University Drive, N.W., Calgary, Alberta T2N lN4, Canada VIII List of Contributors

R. J. P. WILLIAMS, University of Oxford, Inorganic Chemistry Laboratory, South Parks Road, Oxford OX1 3QR, Great Britain M. ZAIDI, Endocrine Unit, Department of Chemical Pathology, Royal Post• graduate Medical School, Hammersmith Hospital, Ducane Road, London W120HS, Great Britain Foreword

The Editorial Board and the Publishers of the Handbook of Experimental Pharmacology wish to express their profound grief at the untimely death of Professor Peter Baker. Aware of his international recognition as an expert on the ubiquitous role of calcium in physiological processes and their pharma• cological control, the Board was gratified when Professor Baker accepted its invitation to edit a new Handbook volume on "Calcium in Drug Actions". He went about this task with his usual energy and effectiveness so that, in the few months before his unexpected death, Professor Baker had mustered his distinguished contributors, got them to provide their manuscripts, and seen almost the entire material into the press. This achievement is all the more remarkable when one bears in mind the extraordinary number of his other commitments during the same time; they are mentioned in Sir Alan Hodgkin's preface to this volume. With so many other professional and personal responsibilities upon him, the Board of the Handbook wishes to record its grateful appreciation for the admirable way in which Professor Baker took on and carried out the additional work of bringing this fine book into existence; and the Board wishes it to be dedicated to the memory of Professor Peter Frederick Baker.

The Editorial Board: G. V. R. BORN, P. CUATRECASAS, H. HERKEN, A. SCHWARTZ PETER FREDERICK BAKER 11 March 1939 to 10 March 1987

PETER BAKER'S sudden death from a heart attack has deprived the international scientific community of one of its most gifted and versatile biologists. He had just finished editing the contributions to this volume of the Handbook of Experimental Pharmacology and it is sad that he will not be here to see the printed book or to take part in the discussion that publication of a major scientific treatise is likely to generate. BAKER'S research activities developed at an early age and probably arose from his interest in natural history. While still at school in Lincoln he published an ecological note on the common earwig and in the interval between school and university he carried out research at Rothamsted which resulted in an article on aphid behaviour on virus-affected sugar beet. After a brilliant undergraduate career at Cambridge, studying Natural Sciences and specialising in Biochemistry in his third year, he decided to take a doctorate in Physiology working on the relationship between phosphorus metabolism and ion transport in nerve. To begin with it was arranged that he would divide his time between Cambridge and Plymouth where there were many interesting experiments to be done with giant nerve fibres that can be obtained from the common squid. XII

At Plymouth BAKER started to work with the late Dr. TREVOR SHAW and almost immediately made a major advance by showing that after the protoplasm had been squeezed out of a giant nerve fibre, conduction of impulses could be restored by perfusing the remaining membrane and sheath with an appropriate solution. With SHAW and the writer, BAKER worked out a method of changing internal solutions while recording with an internal electrode from a perfused axon. It turned out that it did not much matter what internal solution was employed so long as it contained potassium and not much sodium. Provided this condition was satisfied a perfused nerve fibre was able to conduct nearly a million impulses without the intervention of any biochemical processes. ATP is needed for the Na-K pump, but not for the conduction of impulses. There were also some unexpected findings of which one of the most interesting was that reducing the internal ionic strength led to a dramatic shift in the operating characteristics of the membrane. This effect which has a simple physical explanation helps to explain some puzzling findings which were sometimes thought to be inconsistent with the ionic theory of nerve conduction. It did not take PETER BAKER long to finish off the main project he had selected for a Ph.D. thesis, namely the relation of phosphorus metabolism to the active transport of sodium and potassium. In both crab and squid nerve it seems that 3 Na + are ejected per ATP split. The lower figure obtained previously by CALD• WELL, KEYNES, SHAW, and myself was explained by the fact that % of the ATP breakdown occurs in the protoplasm and only )/4 at the membrane. BAKER'S work on the Na-K pump in different tissues led to experiments on the way in which living cells maintain an extremely low level of calcium and the importance of this phenomenon for various kinds of cellular activity. A new finding was the demonstration of a Na-Ca exchange system which helps to maintain internal calcium at a low level at the expense of the sodium gradient. The first evidence for this mechanism which has since been seen in many cells was provided in the autumn and early winter of 1966--67 by BAKER and colleagues working on squid nerve at Plymouth and independently by REUTER and SEITZ at Bern, on heart muscle. PETER BAKER'S interest in calcium was by no means confined to Na-Ca exchange. Some of the many subjects he and his colleagues tackled were the use of aequorin to detect calcium movements in giant nerve fibres, the uncoupled, "Schatzmann" calcium pump, the state of calcium in axoplasm and finally exocytosis. BAKER'S early research was so successful that he soon became a college tutor and university lecturer at Cambridge. He was in fact a brilliant teacher, both in the classroom and as a lecturer. It was, therefore, not surprising that in 1975 he was offered and accepted the Professorship of Physiology at King's College London, where he built up an active centre of research as well as a fine teaching laboratory. At King's College BAKER'S main research interest was in the way in which neural transmitters are discharged by exocytosis. Characteristically, BAKER, and his close colleague, DEREK KNIGHT, chose to attack this fundamental problem in its simplest situation, namely the adrenal medulla, which may be regarded as an extension of the nervous system. Again characteristically, BAKER developed drastic but highly effective ways of getting large molecules into cells. XIII

PETER BAKER was never one to shirk the world's work. He acted as secretary and chairman of the Editorial Board of The Journal of Physiology; was an energetic member of the Medical Research Council's Board on Neurosciences and became a member of the Agricultural and Food Research Council, a post which he greatly enjoyed as he basically felt himself more a biologist than a physiologist. He also served on several Royal Society Committees and one or two University working parties. As if this was not enough he managed to be Chairman of the Parish Council and one of the Governors of the village school at his home in the country in Bourn near Cambridge. In 1966 PETER BAKER married PHYLLIS LIGHT, a geneticist with whom he shared many interests - gardening, natural history and biology among them. They had four children, Lucy, ALEXANDER, SARAH and CHARLOTTE who meant a great deal to him.

A. L. HODGKIN Contents

CHAPTER 1 The Multiple Physiological Roles of Calcium: Possible Sites for Pharmacological Intervention A. W. CUTHBERT 1 References...... 5

Calcium Receptors and Calcium Metabolism

CHAPTER 2 Chemical Factors Determining the Affinity of a Receptor for Calcium B. A. LEVINE and R. J. P. WILLIAMS. With 6 Figures 9 A. Introduction ...... 9 B. Concentrations in Physiological Conditions. 9 C. Calcium-Proton Competition. . . 11 D. The Calcium-Magnesium Problem. 13 E. Other Metal Ions ...... 14 F. Cooperative Interactions. . . . . 14 G. Binding and Conformational Energy 14 H. Kinetic Constraints . . . 16 J. On-Off Binding Constants 16 K. Diffusion of Ions . . . . 17 I. In Water . . . . . 17 II. Through Membranes 18 III. Pumps ...... 19 L. Selective Binding to Unstructured Molecules of High Anionic Charge Density...... 20 M. The Function of Neutral Donors ...... 21 N. Calmodulin: An Example of a Calcium Trigger 23 O. S-100b: An Example ofIon-Ion Cooperativity 26 P. A Look at Phosphorylation Regulation 28 Q. Conclusion. 28 References...... 29 Contents xv

CHAPTER 3 Troponin C and Calmodulin as Calcium Receptors: Mode of Action and Sensitivity to Drugs S. EBASHI and Y. OGAWA. With 4 Figures . . . . . 31 A. Introduction ...... 31 B. Brief Historical Survey of Ca Receptor Proteins . 31 C. General Views of TNC and CaM ...... 33 D. Ca Binding of TNC and CaM ...... 33 E. Ca Binding to TNC in Relation to Tension Development. 37 F. Ca Binding to CaM and Enzyme Activities...... 40 G. CaM Ligands: Their Pharmacologic and Physiologic Significance 43 I. Discovery of CaM-Inhibiting Drugs (CaM Antagonists) . 43 II. CaM Ligands . . 46 H. Concluding Remarks 47 References...... 48

CHAPTER 4 Ligand-Binding Sites on Calmodulin H. J. VOGEL. With 9 Figures 57 A. Introduction . . . . . 57 B. Metal Ion-Binding Sites 59 I. Calcium ..... 59 II. Calcium Probe Cations 62 III. Other Metal Ions 63 C. Binding of Drugs . . . . . 64 I. Trifluoperazine . . . . 64 II. Calmodulin Antagonists 65 III. Calcium Antagonists 66 D. Binding of Peptides . . . . 68 I. Structural Requirements 68 II. Localization of Binding Sites 71 E. Interactions with Target Proteins 72 I. Chemical Modifications and Affinity Labeling 72 II. Activation and Binding with Calmodulin Fragments 74 III. Calmodulin-Binding Domains. 75 F. Epilogue. 76 References...... 80

CHAPTER 5 Calcium Channels as Drug Receptors M. SCHRAMM and R. TOWART. With 5 Figures ...... 89 A. Introduction ...... 89 B. Calcium as a Biological Signalling Mechanism and the Role of Calcium Channels in Maintaining Its Homeostasis...... 90 XVI Contents

C. How Many Types of Calcium Channels Exist? . . . . 91 D. Electrophysiological Properties of the Calcium Channel 93 E. Physiological Modulation of Calcium Channel Function 95 F. Calcium Antagonists ...... 97 G. The 1,4-Dihydropyridine Receptor. 101 H. Calcium Agonists . . . . 104 J. Summary and Conclusions 108 References...... 109

CHAPTER 6 The Chemistry of Calcium Channel Agonists and Antagonists D. J. TRIGGLE. With 13 Figures. 115 A. Introduction ...... 115 B. Structure-Function Studies ...... 116 C. Binding Sites for Ca2+ Channel Ligands 122 D. Ca2 + Channel Binding Sites: Relationship to Ca2+ Channel Function. 125 E. Relationship of Structural and Functional Studies: A Prospective 127 I. Different Categories of Ca2 + Channels...... 127 II. State-Dependent Interactions with Ca2 + Channels. 128 III. Pathologic State of Tissue 128 References...... 129

CHAPTER 7 The Apamin-Sensitive CaB -Dependent K+ Channel: Molecular Properties, Differentiation, Involvement in Muscle Disease, and Endogenous Ligands in Mammalian Brain M. LAZDUNSKI, G. ROMEY, H. SCHMID-ANTOMARCHI, J. F. RENAUD. C. MOURRE, M. HUGUES, and M. FOSSET. With 1 Figure . 135 A. Apamin, Its Structure and Its Active Site...... 135 B. Apamin Blocks Ca2+ -Dependent K + Channels ...... 136 C. The Apamin-Sensitive Ca2+ -Dependent K + Channel is Only One of Several Types of Ca2+ -Dependent K + Channels...... 136 D. Biochemical Properties of the Apamin-Binding Component of the Ca2 + -Dependent K + Conductance ...... 137 E. Apamin as a Tool to Purify the Apamin-Sensitive Ca2+ -Dependent K + Channel and to Determine Its Molecular Weight and Its Polypeptide Composition ...... 138 F. PC12 Pheochromocytoma Cells Hyperproduce the Apamin Receptor and Permit an Analysis of the Internal Ca2+ Concentration Dependence of the Apamin-Sensitive Ca 2 + Channel...... 139 G. Autoradiographic Localization of Apamin-Sensitive Ca2 + -Dependent K + Channels in Rat Brain ...... 140 Contents XVII

H. Developmental Properties of the Ca2+ -Dependent K + Channel in Mammalian Skeletal Muscle and the All-or-None Role of Innervation 141 I. Expression of the Apamin Receptor in Muscles of Patients with Myotonic Muscular Dystrophy ...... 141 J. An Endogenous Apamin-Like Factor Modulating Ca2+ -Dependent K + Channel Activity Exists in Mammalian Brain 142 References...... 143

CHAPTER 8 Drug Effects on Plasma Membrane Calcium Transport F. F. VINCENZI and T. R. HINDS. With 2 Figures 147 A. Introduction ...... 147 B. Calmodulin ...... 148 C. Calmodulin-Binding Site. 151 D. ATPase Catalytic Unit. . 152 E. Phospholipid Environment 152 F. Influx Pathways 154 G. Surface Receptors. 155 H. Summary 157 References...... 158

CHAPTER 9 Development of Inhibitors of Sodium, Calcium Exchange G. J. KACZOROWSKI, M. L. GARCIA, V. F. KING, and R. S. SLAUGHTER. With 4 Figures...... 163 A. Introduction ...... 163 B. Characteristics and Physiologic Properties of Na,Ca Exchange 163 C. Identification of Na,Ca Exchange Inhibitors ...... 167 D. Mechanism of Na,Ca Exchange Inhibition by Amiloride and Bepridil 173 E. Pharmacology of Na,Ca Exchange Inhibitors 178 References...... 180

CHAPTER 10 The Effect of Ruthenium Red and Other Agents on Mitochondrial Calcium Metabolism M. CROMPTON. With 1 Figure ...... 185 A. Introduction ...... 185 B. Inhibitors of the Mitochondrial Ca2 + Transport Systems. 187 I. Ruthenium Red . . . . 187 II. Benzothiazepines. . . . 189 III. Other Ca2+ Antagonists. 189 XVIII Contents

IV. Benzodiazepines 189 V. Trifluoperazine. 189 VI. Gentamicin . . 190 VII. Amiloride Analogues 190 C. Effectors of Ca2+ -Induced Permeabilization 190 D. Mitochondrial Ca2+ Overload ...... 191 I. Mitochondrial Ca2 + and Oxidative Phosphorylation During Ischaemia/Reperfusion ...... 192 II. The Effects of Ruthenium Red and Other Agents on Mitochondrial Ca 2 + 193 References...... 194

CHAPTER 11 Pharmacology of Calcium Uptake and Release from the Sarcoplasmic Reticulum: Sensitivity to Methylxanthines and Ryanodine R. A. CHAPMAN and J. TUNSTALL. With 5 Figures 199 A. Introduction ...... 199 B. Methylxanthines ...... 200 I. The Effect of Caffeine on Ca Uptake . 201 II. The Release of Ca from the Sarcoplasmic Reticulum by Caffeine 201 III. The Effect of Caffeine on the Passive Efflux of Ca...... 205 IV. The Effect of Caffeine on Ca,Mg-ATPase Activity...... 207 V. Heavy and Light Fractions of Isolated Sarcoplasmic Reticulum . 208 VI. Structure-Activity Relations for the Methylxanthines . . 209 C. Ryanodine . 209 D. Conclusions 212 References. . . 213

CHAPTER 12 Effect of Lithium in Stimulus-Response Coupling I. SCHULZ. With 3 Figures 217 A. Introduction ...... 217 B. Lithium ...... 217 C. Phosphoinositides and Calcium 218 D. A Link Between Lithium, Phospholipids and Ca2+ Mobilization 221 E. Inositol-1 ,4,5-Trisphosphate as Second Messenger in the Action of Ca2+ -Mobilizing Hormones . . . . 222 F. Other Inositol Polyphosphates . . . 223 G. Lithium and Manic-Depressive Illness 226 H. Is Lithium a Secretagogue? . . . . 226 J. A Link Between Li +, Phosphoinositides and Cell Proliferation. 229 K. Effects of Li + on Neurotransmitter-cAMP-Stimulated Pathways. 231 L. Conclusion. 232 References...... 233 Contents XIX

CHAPTER 13 Phorbol Esters and Protein Kinase C H. NAGAMOTO, U. KIKKAWA, and Y. NISHIZUKA. With 3 Figures . 241 A. Introduction ...... 241 B. Purification and Assay of Protein Kinase C...... 241 I. Purification of Protein Kinase C from Rat Brain . 241 II. Enzyme Assay of Protein Kinase C . 244 III. Binding Assay of Protein Kinase C . 244 C. Protein Kinase C and Phorbol Esters . . 245 I. Properties ...... 245 II. Biochemical and Physiologic Activation . 245 III. Permeable Diacylglycerol and Phorbol Esters . 246 D. Conclusion. . 249 References...... 249

Calcium and Physiological Function

CHAPTER 14 Drugs Acting on Calcium Channels T. NARAHASHI. With 10 Figures. · 255 A. Introduction . · 255 B. Methods ...... · 256 I. Materials . . . . . · 256 II. Electrical Recording. · 256 III. Solutions . . . . . · 256 C. Two Types of Calcium Channels · 257 I. Initial Study ...... · 257 II. Separation of Two Types of Calcium Channels · 259 III. Kinetics of Two Types of Calcium Channels · 261 IV. Ionic Selectivity...... · 263 V. Sensitivity to Cyclic AMP .. · 263 D. Pharmacology of Calcium Channels · 263 I. Polyvalent Cations · 263 II. Opioid Peptides . · 265 III. Phenytoin . . . . · 269 IV. Pyrethroids. . . . · 270 E. Summary and Conclusions · 271 References...... · 271

CHAPTER 15 Calcium and Synaptic Function M. P. BLAUSTEIN. With 8 Figures ...... 275 XX Contents

A. Introduction ...... 275 B. Regulation of Intracellular Calcium in Nerve Cells...... 276 I. The Intracellular Free Calcium Concentration in Nerve Cells 276 II. Calcium Entry into Nerve Cells...... 276 III. Intracellular Calcium Buffering in Nerve Cells 279 1. Mitochondria ...... 279 2. Smooth Endoplasmic Reticulum . . . . . 279 3. Cytosolic Buffers...... 282 IV. Calcium Transport Across the Neuronal Plasma Membrane. 283 V. The "Life Cycle" of Calcium at the Nerve Terminal . . 289 C. The Role of Intracellular Calcium in Synaptic Transmission 289 I. Calcium and Neurotransmitter Release...... 290 II. Dissection of the Steps in Transmitter Release with Toxins and Drugs ...... 292 III. Calcium and the Control of Excitability in Neurons 293 IV. Calcium and Memory 295 D. Summary and Conclusions 296 References...... 297

CHAPTER 16 Some New Questions Concerning the Role of Cal + in Exocytosis S. COCKCROFT and B. D. GOMPERTS. With 6 Figures . . . . . 305 A. Calcium and Cell Activation ...... 305 B. Exocytosis as an Example of a Cellular Activation Process 309 I. Adrenal Chromaffin Cells 310 II. Neutrophils ...... 310 III. The Mast Cells ...... 311 C. Direct Manipulation of Cytosol Ca2+ 312 I. Calcium Ionophores...... 312 II. Manipulation of Cytosol Ca2+ in Permeabilised Cells 313 III. Methods of Plasma Membrane Permeabilisation. . . 313 IV. Ca2+ -Induced Secretion from Permeabilised Neutrophils 314 V. Ca2 + -Induced Secretion from Permeabilised Adrenal Chromaffin Cells ...... 315 D. Exocytotic Secretion Without Elevation of Cytosol Ca2 + 318 I. Phorbol Ester ...... 318 II. Guanine Nucleotides ...... 318 E. A Role for G-Protein in Exocytosis? ...... 319 F. Questions Concerning the Role of Ca2+ in Exocytosis 322 I. Single Cells...... 322 1. Membrane Capacitance Changes in Exocytosis 322 2. Fast Ca2+ Transients Are not Sufficient to Trigger Exocytosis. 325 G. Towards Reconstitution of Exocytosis in Cell-Free Systems . 327 References...... 329 Contents XXI

CHAPTER 17 Exo-Endocytosis: Mechanisms of Drug and Toxin Action J. MELDOLESI, T. POZZAN, and B. CECCARELLI. With 3 Figures 339 A. Introduction ...... 339 B. Exocytosis ...... 340 I. Second Messenger Control of Regulated Exocytosis 342 II. Membrane Fusion-Fission in Exocytosis 345 III. Drugs ...... 347 IV. Toxins...... 349 1. Toxins Targeted to Channels and Receptors 349 2. Clostridium Toxins: Inhibitors of Exocytosis 349 3. (J(-Latrotoxin and Congeners: Stimulators of Exocytosis 351 C. Endocytosis ...... 352 I. Membrane Sorting in Endocytosis . 352 II. Regulation of Endocytosis 354 References...... 355

CHAPTER 18 Pharmacology of Calcium Metabolism in Smooth Muscle T. GODFRAIND. With 11 Figures ...... 361 A. Introduction ...... 361 B. Calcium Entry and Calcium Regulation at Rest: Action of Pharmacologic Agents...... 361 C. Calcium Movements During Excitation: Their Sensitivity to Pharmacologic Agents...... 366 D. Heterogeneity of Excitation-Contraction Coupling Mechanisms 370 E. A Pharmacologic Example: Contraction of Vascular Smooth Muscle, Role of Endothelium, and Action of Dihydropyridines and Diphenylpiperazines. . . . 372 I. Inhibition of Contraction 373 II. Role of Endothelium . 377 F. Concluding Remarks 380 References...... 380

CHAPTER 19 Drugs Affecting Cardiac Calcium Metabolism P. HONERJAGER. With 2 Figures. 383 A. Introduction ...... 383 B. The Calcium Signal ...... 383 C. Classification and Selection of Drugs 386 D. Drugs Affecting Sarcolemmal Calcium-Transporting Proteins 393 I. Calcium Channels ...... 393 1. Direct Inhibitors and Activators: "Calcium Antagonists" and "Calcium Agonists" ...... 393 XXII Contents

2. Drugs Acting via Cyclic AMP. · 396 3. Other Drugs . . . . . · 398 II. Sodium/Calcium Exchange . . . · 399 1. Direct Inhibitors ...... · 399 2. Drugs Affecting Intracellular Sodium · 399 E. Drugs Affecting Sarcoreticular Calcium-Transporting Proteins · 401 F. Drugs Affecting Mitochondrial Calcium-Transporting Proteins · 402 G. Conclusions · 403 References...... · 403

CHAPTER 20 Hormonal Control of Extracellular Calcium I. MACINTYRE, M. ZAIDI, C. MILET, and P.J.R. BEVIS. With 17 Figures. 411 A. Introduction . . . . . 411 B. Parathyroid Hormone . 411 I. Chemistry . . . 411 II. Biosynthesis. . . 411 III. Secretion and Metabolism. . 412 IV. Biological Actions . . . . . 414 V. Pathophysiology...... 415 1. Primary Hyperparathyroidism. . 415 2. Renal Failure...... 416 C. Hormone from the Corpuscle of Stannius . 416 I. Chemistry and Biosynthesis . 416 II. Biological Action . . . . . 416 D. Vitamin D: Endocrine System. . . 417 I. Chemistry and Biosynthesis . 417 II. Metabolism...... 417 1. Mineral Regulation . . . 419 2. Hormonal Regulation . . 419 a) Parathyroid Hormone. . 419 b) Growth Hormone and Prolactin . 419 c) Calcitonin...... 420 d) Vitamin D Metabolites . 420 III. Biological Actions . . . . 420 IV. Pathophysiolog~. . . . . 420 E. The Calcitonin Gene Peptides . . 422 I. Discovery. . . 422 1. Calcitonin ...... 422 2. Katacalcin ...... 422 3. Calcitonin Gene-Related Peptide . 422 II. Biosynthesis. . 422 III. Chemistry . 423 IV. Secretion . 426 V. Actions. . . 428 Contents XXIII

1. Calcitonin ...... 428 2. Calcitonin Gene-Related Peptide . 430 VI. Physiological Role...... 431 1. Calcitonin ...... 431 2. Calcitonin Gene-Related Peptide . 432 VII. Pathophysiology: Medullary Carcinoma of the Thyroid . 433 VIII. Therapeutic Considerations . 433 1. Calcitonin . . . . . 433 a) Paget's Disease. . 433 b) Osteoporosis. . . 434 c) Hypercalcaemia . 434 2. Calcitonin Gene-Related Peptide . 434 References...... 434

CHAPTER 21 Bisphosphonates: A New Class of Drugs in Diseases of Bone and Calcium Metabolism H. FLEISCH ...... 441 A. Introduction ...... 441 B. Chemistry and General Characteristics. . 441 C. Synthesis ...... 443 D. Methods of Determination. . 443 E. Monophosphonates . . 444 F. History ...... 444 G. Mode of Action ...... 445 I. Physicochemical Effects . 445 II. Effect on Calcification In Vivo . 445 III. Inhibition of Bone Resorption . 446 IV. Biochemical and Cellular Effects . 448 V. Mode of Action in the Inhibition of Bone Resorption . 449 VI. Other Effects . . 450 H. Pharmacokinetics. . 451 J. Toxicology. . . . . 451 K. Drug Interactions . . 452 L. Clinical Use . . . . 452 I. Ectopic Calcification and Ossification . 452 1. Soft Tissue Calcification. . 452 2. Urolithiasis ...... 452 3. Dental Calculus ...... 453 4. Fibrodysplasia Ossificans Progressiva . 453 5. Other Heterotopic Ossifications . . . 453 II. Abnormally Increased Bone Resorption . 454 1. Paget's Disease...... 454 2. Primary Hyperparathyroidism . . . . 455 3. Hypercalcemia of Malignancy and Tumoral Bone Destruction. 455 4. Osteoporosis...... 456 XXIV Contents

M. Side Effects · 456 N. Future Prospects · 457 References. . . . . · 457

CHAPTER 22 Calcium and Hypertension R. D. BUKOWSKI and D. A. MCCARRON. With 2 Figures . 467 A. Introduction ...... 467 B. Role of Ca2+ in Vascular Smooth Muscle Contraction. . 468 C. Role of Altered Ca2+ Metabolism in Hypertension . 470 I. Human Studies...... 470 II. The Spontaneously Hypertensive Rat . 471 III. Ca2+ Antagonists and Hypertension. . 472 D. Role of Dietary Calcium in Hypertension . 473 I. Human Studies ...... 473 II. The Spontaneously Hypertensive Rat . 474 E. Postulated Mechanisms ...... 476 I. Effect of Dietary Calcium on Vascular Smooth Muscle. . 476 II. Dietary Calcium and Phosphate Metabolism ...... 479 III. Dietary Calcium, Sodium Metabolism, and Fluid Balance . 479 IV. Dietary Calcium and the Autonomic Nervous System . 480 V. Composite Hypothesis. . 481 F. Conclusions . 482 References...... 482

Drugs and Toxicological Agents that Either Mimic Calcium or Elements of Intracellular Calcium Metabolism

CHAPTER 23 Calcium Chelators and Calcium Ionophores D. M. BERS and K. T. MACLEOD. With 3 Figures . 491 A. Introduction . . . . 491 B. Calcium Chelators ...... 491 I. EGTA...... 491 II. Carboxylate Ca2+ Chelators and Fluorescent Ca2+ Probes . 495 III. Metallochromic Dyes . 497 IV. Photoproteins ...... 498 C. Calcium Ionophores...... 498 I. Naturally Occurring Calcium Ionophores . . 499 1. X-537 A . . 500 2. A23187 . . 501 3. Ionomycin. 503 Contents xxv

II. Synthetically Produced Calcium Ionophores 503 1. DDP- and DOPP- . 503 2. ETH 1001 . 504 D. Conclusion. 504 References...... 505

CHAPTER 24 Lead-Calcium Interactions and Lead Toxicity T. J. B. SIMONS ...... 509 A. Overview ...... 509 B. Relevant Chemistry of Lead and Calcium 509 I. Introduction ...... 509 II. Chemistry of the Ions in Solution . . 510 1. Complexes with Simple Anions. . 510 2. Complexes with Organic Ligands . 510 3. Ionic Radius...... 511 III. Measurement of Pb2 + Concentration 511 IV. Pb2 + Buffers...... 511 C. Nonenzymic Actions of Lead...... 512 D. Interactions Between Lead and Binding Proteins 512 I. Calmodulin ...... 512 II. Troponin C ...... 512 III. Intestinal Calcium-Binding Proteins 513 IV. Lead-Binding Proteins. 513 V. Summary ..... 514 E. Lead-Enzyme Interactions . 514 I. Ca2+ -ATPase .... 514 II. b-Aminolevulinic Acid Dehydratase 514 III. Adenylate Cyclase...... 515 IV. Na+, K+-ATPase ...... 515 V. Calmodulin-Dependent Actions. 515 VI. Summary ...... 515 F. Transport of Lead, and Its Effects Upon Ion Transport 516 I. Transport Across the Plasma Membrane . 516 1. Human Red Blood Cells. 516 2. Ca Channels...... 517 II. Transport Across Epithelia . 517 1. Intestinal Absorption . . 517 2. Renal Absorption and Excretion 518 III. Mitochondria...... 518 G. Cellular Homeostasis ...... 519 H. Neurotransmission and Neurosecretion. 520 J. Summary and Conclusions 521 References...... 522 XXVI Contents

CHAPTER 25 Alkaline Earths, Transition Metals, and Lanthanides C. H. EVANS. With 5 Figures...... 527 A. Introduction ...... 527 B. Interactions of Alkaline Earths, Transition Metals, and Lanthanides with Calcium Channels ...... 527 I. Slow, Voltage-Operated Calcium Channels ...... 528 II. Other Types of Calcium Transport Mechanisms ...... 532 C. Cellular Physiologic Effects of Inorganic Blockers of Calcium Channels 533 D. Metal Ions as Drugs...... 536 E. Metabolism of Lanthanides by Whole Animals 536 F. Possible Therapeutic Uses of the Lanthanides . 539 G. Summary 542 References. . 543

Subject Index 547