Angelo Azzi Zdenek Drahota Sergio Papa (Eds.)

Molecular Basis of Membrane-Associated Diseases

With 121 Figures

Springer-Verlag Berlin Heidelberg New York London Paris Tokyo Hong Kong Prof. Dr. ANGELO AZZI Institut für Biochemie und Molekularbiologie Universität Bern Bühlstraße 28 CH-3012 Bern

Prof. ZDENEK DRAHOTA Institute of Physiology Czechoslovak Academy of Sciences Videnska 1083 CS-142 20 Praha 4

Prof. SERGIO PAPA Institute of Medical Biochemistry and Chemistry University of Bari Piazza G. Cesare 1-70124 Bari

ISBN 3-540-50744-2 Springer-Verlag Berlin Heidelberg New York ISBN 0-387-50744-2 Springer-Verlag New York Berlin Heidelberg

Library of Congress Cataloging-in-Publication Data. Molecular basis of membrane-associated diseases. Based on a symposium held in Prague on 14-16 July 1988. Includes index. 1. Cell membranes-Abnormalities- Congresses. 2. Mitochondrial membrancs-Abnormalitics-Congresses. 3. Lysosomal storage diseases-Congres• ses. I. Azzi. A. (Angelo) II. Drahota, Ζ. III. Papa. S. [DNLM: 1. Cell Membranc-pathology-congresses. 2. Diseases-ctiology-congresses. 3. Membranes-pathology-congresscs. QS 532.5.M3 M718 1988] RB152.M64 1989 616.07 89-11270

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© Springer-Verlag Berlin Heidelberg 1989 Printed in Germany

The use of registered names, trademarks, etc. in this publication docs 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.

Typesetting: International Typesetters Inc. Makati, Philippines Printing: Druckhaus Beltz, Hemsbach Binding: J. Schäffer, Grünstadt 2131/3145-543210- Printed on acid-free paper Contents

1 Plasma Membrane and Control of Cell Growth and Differentiation

Insulin Receptor Kinase and Insulin Action Ε. VAN OBBERGHEN, R. BALLOTTI, Y. LE MARCHAND-BRUSTEL and J. C. SCIMECA (With 3 Figures) 3 Structural and Functional Aspects of Signal Transduction by Receptor Tyrosine Kinases. Y. YARDEN 14 Nerve Growth Factor (NGF): Physiological Functions and Regulation of Its Synthesis. D. LINDHOLM, R. HEUMANN, G.D. BORASIO and H. THOENEN 22 Signal Transduction by Growth Factor Receptors W. H. MOOLENAAR (With 3 Figures) 29 Altered Regulation of Receptor-Associated Tyrosine Kinases in Human Tumors. RM. COMOGLIO, M.F. DI RENZO, G. GAUDINO, S. GIORDANO, L. NALDINI and C. PONZETTO 35 The Role of Plasma Membrane Phospholipids in Cellular Pathology A. STRUNECKA, J. PALECEK, L. KRPEJSOVA, N.I. EL DESOUKI, P. FOLK and J. MACHA (With 7 Figures) 53 Changes in the Structure and Function of the Red Cell Membrane Skeleton and Hemolytic Anemias. P. JAROLIM, S.-C. LIU, TH. L. COETZER, M. LAHAV and J. PALEK (With 1 Figure) 64

Regulation of Intracellular Ca2+ in Pancreatic Acinar Cells by Membrane Pumps and Channels. R.L. DORMER (With 3 Figures) .... 71 Control of Protein Kinase C Function by Activators and Inhibitors CH. W. MAHONEY and A. Azzi (With 2 Figures) 84

2 Regulation of Membrane Channels

K+ Channels: Structure, Function, Regulation, Molecular Pharmacology and Role in Diseased States. H. BERNARDI, J.-N. BIDARD, M. FOSSET, M. HUGUES, CH. MOURRE, H. REHM, G. ROMEY, H. SCHMID- ANTOMARCHI, H. SCHWEITZ, J. R. DE WEILLE and M. LAZDUNSKI (With 9 Figures) 99 VIII Contents

Voltage-Dependent Calcium Channels: Structure and Regulation in Normal and Abnormal States. M.M. HOSEY, F. C. CHANG, CM. O'CALLAHAN, J. PTASIENSKI, J. BARHANIN and M. LAZDUNSKI (With 6 Figures) 112 Cell Membrane Abnormalities in Genetic Hypertension G. BIANCHI, P. FERRARI, S. SALARDI, P. SALVATI, M. FERRANDI, L. TORIELLI and B.R. BARBER 124

Transmembrane Cation Transport: An Approach to the Study of the Molecular Basis of Hypertension. L. MAZZANTI, R. A. RABINI, N. CESTER, C. ROMANINI and E. BERTOLI 130

Transmembrane Ionic Fluxes in the Brain in Energy-Depleted Cells Β. K. SIESJÖ (With 9 Figures) 134

3 Mitochondrial Genetic Diseases

Mitochondrial Diseases. S. DIMAURO, A. LOMBES, H. NAKASE, M. ZEVIANI, R. RIZZUTO, C.T MORAES and E. A. SCHON 157

Mitochondrial Myopathy: Biochemical Approaches to Respiratory Chain-Linked Electron Transfer and Energy Coupling CP LEE, M.E. MARTENS, PL. PETERSON, S.H. TSANG and M. SCIAMANNA (With 6 Figures) 167

Phosphorus Magnetic Resonance Spectroscopy (31P NMR) as a Tool for in Vivo Monitoring of Mitochondrial Muscle Disorders Z. ARGOV (With 5 Figures) 183 Cytochemical and Immunocytochemical Studies of Human Mitochondrial Myopathies. M. A. JOHNSON (With 7 Figures) 200

Regulatory Complexity of Cytochrome c Oxidase and Its Defective Manifestation in Mitochondrial Diseases. B. KADENBACH, F.-J. HÜTHER, U. BÜGE, A. SCHLERF and M. A. JOHNSON (With 7 Figures) 216

Fibroblasts and Cytochrome c Oxidase Deficiency D.M. GLERUM, B.H. ROBINSON and R. CAPALDI (With 2 Figures) . . . 228

Human Cytochrome c Oxidase Deficiencies; Structural and Functional Aspects. K.M.C SINJORGO, A.O. MUIJSERS, H.R. SCHÖLTE,

Α. Β. P. VAN KUILENBURG, J.J. VAN BEEUMEN, E.M. BROUWER-KELDER, J. A. BERDEN, P.A. BOLHUIS and J.M. TAGER (With 3 Figures) 239

4 Mitochondrial Metabolic Diseases

Brown Adipose Tissue, Diet-Induced Thermogenesis and Genetic Obesity. T. SOUKUP, H. RAUCHOVÄ, J. DOSOUDILOVA and Z. DRAHOTA (With 5 Figures) 257 Contents IX

Acute and Long-Term Regulation of Brown Adipose Tissue Thermogenesis: Physiological and Pathological Implications J. HOUSTEK, J. KOPECKY, D. JANI'KOVÄ, J. BEDNAR, Μ. HOLUB, T. SOUKUP, M. MIKOVA and K. VI'ZEK (With 7 Figures) 265

Molecular Composition, Fluidity of Membranes and Functional Properties of Human Liver Mitochondria and Microsomes GH. BENGA (With 3 Figures) 285 Changes in Erythrocyte and Platelet Membrane Fluidity in Childhood Obesity. G. FERRETTI, A. TANGORRA, PL. GIORGI, M. DOTTI, G. CURATOLA and E. BERTOLI 303

Skeletal Muscle Mitochondria and Phospholipase Activity in Malignant Hyperthermia. K.S. CHE AH 309 The Antimitochondrial Antibodies (ΑΜΑ) of Primary Biliary Cirrhosis (PBC). H. BAUM and M. FUSCONI 323

Toxicity, Antioxidants, and Metabolism. L. PACKER (With 3 Figures) . . 335

5 Cancer and Energy Metabolism

Mitochondrial H+-ATP Synthase Under Normal and Pathological Conditions. S. PAPA, F. GUERRIERI, F. ZANOTTI and F. CAPUANO (With 8 Figures) 347 Antitumor Effect of Drugs Interfering with Mitochondrial Biogenesis S. KUZELA, K. LUCIAKOVA, J. KOLAROV and B.D. NELSON (With 6 Figures) 359

6 Lysosomes and Peroxisomes in Health and Disease

The Complex of /3-Galactosidase, Neuraminidase and "Protective Protein" in Lysosomes: Molecular Characterization of the "Protective Protein". A. D'AZZO, N. GILLEMANS and N. GALJART (With 5 Figures) . 371

The Role of Activator Proteins in Glycolipid Degradation and Storage Diseases. E. CONZELMANN (With 3 Figures) 379 Solute Translocation Across the Lysosome Membrane: Physiology, Pathology and Pharmacology. J.B. LLOYD, T. KOOISTRA, S. FORSTER, S.J. BIRD and G.P. IVESON (With 6 Figures) 395 The Peroxisomal /^-Oxidation Systems: Characteristics and (Dys) Functions in Man. R.J.A. WANDERS, C.W.T. VAN ROERMUND, Μ. J. A. VAN WIJLAND, R.B.H. SCHUTGENS, H. VAN DEN BOSCH and J.M. TAGER 407 χ Contents

Genetic Heterogeneity in Inherited Disorders with a Generalized Impairment of Peroxisomal Functions: Visualization by Immunofluorescence Microscopy of Peroxisome Assembly After Somatic Cell Fusion of Complementary Cell Lines S. BRUL, E. A.C. WIEMER, M. OOSTHUIZEN, A. WESTERVELD, R. VAN DRIEL, A. STRIJLAND, R.B.H. SCHUTGENS, Η. VAN DEN BOSCH, R. J. A. WANDERS, A. W. SCHRAM and J. M. TAGER (With 2 Figures) . . 420

Subject Index 429 List of Contributors

You will find the addresses at the beginning of the respective contribution

Argov, Ζ 183 ElDesouki, N.I. 53 Azzi, A. 84 Ferrandi, M. 124 Ballotti, R. 3 Barber, B.R. 124 Ferrari, P. 124 Barhanin, J. 112 Ferretti, G. 303 Baum, Η. 323 Folk, P. 53 Bednar, J. 265 Forster, S. 395 Benga, Gh. 285 Fosset, M. 99 Berden, J.A. 239 Fusconi, M. 323 Bernardi, H. 99 Bertoli, E. 130, 303 Galjart, N. 371 Bianchi, G. 124 Gaudino, G. 35 Bidard, J.-N. 99 Gillemans, N. 371 Bird, S.J. 395 Giordano, S. 35 Bolhuis, P.A. 239 Giorgi, P.L. 303 Borasio, G.D. 22 Glerum, D.M. 228 Brouwer-Kelder, Ε. M. 239 Guerrieri, F. 347 Brul, S. 420 Büge, U. 216 Heumann, R. 22 Holub, M. 265 Capaldi, R. 228 Hosey, M.M. 112 Capuano, F. 347 Houstek, J. 265 Cester, N. 130 Hugues, M. 99 Chang, F.C. 112 Hüther, F.-J. 216 Cheah, K.S. 309 Coetzer, Th. L. 64 Iveson, G.P. 395 Comoglio, P.M. 35 Conzelmann, E. 379 Janikovä, D. 265 Curatola, G. 303 Jarolim, P. 64 d'Azzo, A. 371 Johnson, M. A. 200,216 de Weille, J.R. 99 DiMauro, S. 157 Kadenbach, Β. 216 DiRenzo, M.F. 35 Kolarov, J. 359 Dormer, R.L. 71 Kooistra, T. 395 Dosoudilovä, J. 257 Kopecky, J. 265 Dotti, M. 303 Krpejsovä, L. 53 Drahota, Z. 257 Kuzela, S. 359 XII List of Contributors

Lahav, M. 64 Salardi, S. 124 Lazdunski, M. 99, 112 Salvati, P. 124 Lee, CP. 167 Schlerf, A. 216 Le Marchand-Brustel, Y. 3 Schmid-Antomarchi, H. 99 Lindholm, D. 22 Schölte, H.R. 239 Liu, S.-C. 64 Schon, E. A. 157 Lloyd, J. B. 395 Schräm, A.W. 420 Lombes, A. 157 Schutgens, R.Β. Η. 407, 420 Luciakovä, K. 359 Schweitz, Η. 99 Sciamanna, Μ. 167 Mächa, J. 53 Scimeca, J.C 3 Mahoney, Ch. W. 84 Siesjö,B.K. 134 Martens, M.E. 167 Sinjorgo, K.M.C 239 Mazzanti, L. 130 Soukup, T. 257, 265 Mikovä, M. 265 Strijland, A. 420 Moolenaar, W.H. 29 Struneckä, A. 53 Moraes, CT. 157 Mourre, Ch. 99 Tager, J.M. 239,407,420 Muijsers, A.O. 239 Tangorra, A. 303 Thoenen,H. 22 Nakase, H. 157 Torielli, L. 124 Naldini, L. 35 Tsang, S.H. 167 Nelson, B.D. 359

O'Callahan, CM. 112 van Beeumen, J.J. 239 Oosthuizen, M. 420 van den Bosch, H. 407, 420 van Driel, R. 420 Packer, L. 335 van Kuilenburg, A. B. P. 239 Palecek, J. 53 Van Obberghen, E. 3 Palek, J. 64 van Roermund, C W. T. 407 Papa, S. 347 van Wijland, M.J.A. 407 Peterson, P. L. 167 Vizek, K. 265 Ponzetto, C 35 Ptasienski, J. 112 Wanders, R.J.A. 407, 420 Westerveld, A. 420 Rabini, R.A. 130 Wiemer, E.A.C. 420 Rauchovä, H. 257 Rehm, H. 99 Rizzuto, R. 157 Yarden, Y. 14 Robinson, Β. Η. 228 Romanini, C. 130 Zanotti, F. 347 Romey, G. 99 Zeviani, M. 157 Nerve Growth Factor (NGF): Physiological Functions and Regulation of Its Synthesis

D. LINDHOLM1, R. HEUMANN, G. D. BORASIO and H. THOENEN

1 Introduction

The physiological role of NGF as a neurotrophic factor in the peripheral nervous system is well established. NGF is essential for the development and maintenance of specific functions of peripheral sympathetic and neural-crest derived sensory neurons. NGF acts as a retrograde messenger between the target organs and the innervating neurons. The regionally different quantities of NGF in target tissues reflect the density of innervation by NGF-responsive neurons, which transport NGF in membrane-confined compartments to the perikaryon after NGF is bound via specific membrane receptors. Although the physiological function of NGF is well documented and a large number of NGF-specific effects on target neurons have been identified, there are two main questions which remain to be elucidated, namely the nature of the second messenger(s) evolving from the binding of NGF to its receptor and the mode of regulation of NGF synthesis in the target tissues. A better understanding of this regulation would provide the prerequisite for a rational approach to the pharmacological modification of NGF synthesis. This is of particular interest because it has recently become apparent that NGF is essential for the preservation of the function of the cholinergic neurons of the basal forebrain nuclei. These neurons are consistently affected in Alzheimers's disease and their impaired function seems to be largely responsible for the cognitive deficits of this disease.

2 Physiological Functions of NGF

Earlier work on NGF was mainly focussed on the role of NGF in embryonic development and differentiation of the peripheral nervous system (Levi-Mon- talcini 1987). NGF has been shown to regulate the survival of peripheral sym• pathetic and neural-crest derived sensory neurons during restricted periods of development (cf. Levi-Montalcini and Angeletti 1968; Greene and Shooter 1980; Thoenen and Barde 1980). This has been conclusively demonstrated in exper• iments on neonatal rodents where administration of anti-NGF antibodies leads to

Department of Neurochemistry, Max-Planck-Institute for Psychiatry, Am Klopferspitz 18 A,D-8033 Martinsried/Munich, FRG 'To whom correspondence should be addressed

A. Azzi, Z. Drahota and S. Papa Molecular Basis of Membrane-Associated Diseases © Springer-Verlag Berlin Heidelberg 1989 Nerve Growth Factor 23 enhanced cell death in the NGF-responsive neuronal population, whereas ex• ogenous NGF can rescue neurons which normally would die during the period of natural cell death. In addition, NGF is also required for differentiation and maintenance of function of the peripheral sympathetic and sensory neurons in adult life. Thus, NGF has been shown to regulate synthesis of specific enzymes which are involved in neurotransmitter production, such as tyrosine hydroxylase and dopamine /^-hydroxylase as well as a series of neuron-specific peptides (cf. Otten 1984; Johnson E.M. et al. 1986; Thoenenetal. 1987). In the peripheral target fields NGF is synthesized in very low amounts and is retrogradely transported to the cell bodies of NGF-responsive neurons after being taken up via specific cell surface receptors (cf. Schwab et al. 1982). It has also been shown that the density of innervation of target organs by NGF-responsive neurons correlates with the levels of NGF and its mRNA (Korsching and Thoenen 1983; Heumannetal. 1984; Shelton and Reichardt 1984). Moreover, interruption of the retrograde axonal transport of NGF following axotomy or by pharmacological interference (e.g. colchicine treatment which results in disassembly of microtubuli) leads to degenerative changes in the NGF-responsive neurons, demonstrating that NGF acts as a retrograde messenger between the periphery and the innervating neurons (cf.Thoenen etal. 1987). In contrast to sympathetic neurons in the peripheral nervous system, central adrenergic neurons neither respond to NGF nor carry NGF receptors (Konkol et al. 1978; Schwab et al. 1979). However, evidence for a trophic role of NGF in the central nervous system became apparent when it was shown that the cholinergic neurons of the basal forebrain nuclei express NGF receptors and exhibit specific retrograde transport of NGF (Schwab et al. 1979; Seiler and Schwab 1984). The cholinergic neurons have also been shown to respond to NGF with an increase in the levels of choline acetyltransferase (ChAT), the enzyme responsible for synthesis of the cholinergic neurotransmitter acetylcholine. This has been shown both in neonatal rats (Gnahn et al. 1983), in aggregate cultures of fetal forebrain (Honegger and Lenoir 1982), and in adult rats after partial fimbria lesions, i.e. of the septo-hippocampal connection (Hefti et al. 1984). The cell bodies of the cholinergic neurons are situated in the medial septal nuclei, in the diagonal band of Broca and in the basal nuclei of Meynert. Their fiber tracts project to the hippocampus, neocortex and olfactory bulb (cf. Cuello and Sofroniew 1984) and these innervated areas exhibit relatively high levels of NGF and NGF-mRNA as compared to other brain regions (Korsching et al. 1985; Whittemore et al. 1986; Shelton and Reichardt 1986). In analogy to the situation in the peripheral nervous system, there is a close correlation between the levels of NGF and its mRNA and the density of cholinergic innervation. Even within the rat hippocampus there are differences between NGF levels of various regions, e.g. the CA3, CA4 and dentate gyrus region have higher levels of NGF protein compared with the CA1-CA2 region and this is reflected in the density of cholinergic innervation (cf. Korsching 1986; Thoenen et al. 1987). Recent autoradiographic receptor binding studies on sections from adult rat brain have clearly demonstrated the presence of NGF receptors on these cholinergic neurons (Riopelle et al. 1987; Raivich and Kreutzberg 1987). Moreover, NGF levels and ChAT activity in rat hippocampus 24 D. Lindholm et al. increase in parallel during postnatal development, further supporting a role of NGF in this septo-hippocampal system (Auburger et al. 1987). Likewise, fimbria lesion experiments have shown that exogenous NGF administrated through an infusion pump can rescue cholinergic neurons of the basal forebrain nuclei which would otherwise degenerate following the transection (Hefti 1986; Kromer 1987; Williamset al. 1986).

3 Mechanism(s) of Action of NGF

Although many actions of NGF on target neurons have been described, the overall mechanism by which NGF exerts its effects is still unknown. This also holds true for the most important long-term effect of NGF, namely its survival-promoting activity on NGF-responsive neurons. NGF receptors on sensory (Sutter et al. 1979) and sympathetic neurons (Godfrey and Shooter 1986) and on PC 12 pheo- chromocytoma cells (Hosang and Shooter 1985) have been identified and antibodies to the NGF receptor have been produced (Chandler et al. 1984; Ross et al. 1984). Moreover, binding studies have shown the existence of both high- and low-affinity NGF receptors on target cells which are encoded by the same mRNA. However, the mechanism for conversion between the two forms is not known but the high-affinity receptor seems to be responsible for most of the biological action of NGF, at least in PC12 cells (Green et al. 1986). Recently, both human and rat NGF receptors have been cloned (Johnson D. et al. 1986; Radeke et al. 1987) and the molecular data demonstrate that the NGF receptor belongs to a novel class of membrane receptors which lack an intrinsic tyrosine kinase activity. NGF is also known not to affect neuronal proliferation which is in contrast to most growth factors which have a mitogenic activity on their target cells (cf. Thoenen et al. 1987). Previous studies have shown that intracellular microinjection of NGF into the cytoplasm of PC 12 cells, bypassing the receptor, fails to elicit differentiation which is readily seen with NGF in the culture medium (Heumann et al. 1981). Conversely, microinjected, function-blocking, anti-NGF antibodies do not inhibit NGF action on PC 12 cells. These studies led to the conclusion that NGF acts by a distinct second messenger system, the nature of which remains to be elucidated (Heumann et al. 1981). The results of recent studies have suggested a role for the ras oncogene product in NGF signal transduction. Introduction of purified, activated, ras protein into PC 12 cells can elicit differentiation (Bar-Sagi and Feramisco 1985), and microinjected, anti-ras antibodies are able to block the effect of NGF on these cells (Hagag et al. 1986). The exact site of action of ras or of ras-like proteins in the signal transduction cascade induced by NGF is not known. Further downstream, transcriptional activators are most probably involved in regulating NGF-specific gene expression. Interestingly, NGF (Greenberg et al. 1985) as well as ras protein (Stacey et al. 1987) are known to transiently increase expression of the c-fos proto-oncogene product which is thought to be involved in transcriptional regulation. Milbrandt (1987) has recently described another putative transcrip• tional activator gene whose transcription is strongly induced in PC 12 cells by short-term NGF treatment. Nerve Growth Factor 25

4 Regulation of NGF Synthesis and Implications for Treatment of Alzheimer's Disease

In Alzheimer's disease the cholinergic neurons of the basal forebrain nuclei are consistently affected and there is an atrophy and /or loss of these neurons during the course of this disease (Coyle et al. 1983; Pearson et al. 1983). Moreover, the impaired synthesis of acetylcholine is one of the earliest detectable signs of Alzheimer's disease and correlates well with the cognitive deficits observed (Francis et al. 1985). In view of the fact that these cholinergic neurons are also the target of NGF in the central nervous system, the hypothesis was put forward that NGF could play a role in this disease (Hefti and Weiner 1986). The idea that Alzheimer's disease results from an insufficient supply of NGF for these neurons has recently been disputed by Goedert and co-workers (1986). These authors demonstrated that the levels of NGF-mRN A in the cerebral cortex of patients with Alzheimer's disease are not different from the corresponding levels found in age-matched controls. Nevertheless, the following observations support the notion that pharmacological amounts of NGF are of potential value in treatment of Alzheimer's disease by supporting the cholinergic neurons undergoing atrophy or degeneration. Thus, it has been demonstrated that chronic administration of NGF over a period of 4 weeks can improve memory function, as shown by behavioural tests, and can counteract cholinergic cell body atrophy observed in a subpopulation of aged rats (Fischer et al. 1987). Moreover, studies on septo-hippocampal lesions clearly show that exogenous NGF has a beneficial effect in preventing cholinergic cell degeneration in the septal area after a transection of the fimbria (Hefti 1986; Kromer 1987; Williams et al. 1986). Likewise, NGF injections (Will and Hefti 1985) or transplantation of fetal cholinergic neurons into the hippocampus (Dunnett et al. 1982; Gage and Björklund 1986) can improve both learning and memory deficits observed in rats after such a lesion. In view of these findings it seems justified to consider NGF supplementation as a possible therapeutic approach for the treatment of patients suffering from Alzheimer's disease. This could be achieved by either local administration of recombinant human NGF, of peptides with NGF-like functions or by enhancing endogenous NGF synthesis by pharmacological agents. However, a prerequisite for such an intervention is a better understanding of the physiological mechanisms by which NGF is regulated in vivo. Recent studies in the peripheral nervous system have shown that specific polypeptide factors are involved in the control of NGF synthesis in the nonneuronal cells. Using the rat sciatic nerve as a model system under conditions of degeneration and regeneration after a nerve lesion, we were able to show that macrophages play an important role in NGF regulation (Heumann et al. 1987; Thoenen et al. 1988). Interleukin-1, a lymphokine produced by these cells, was found to be the responsible agent increasing NGF-mRN A levels in the nonneuronal cells (Lindholm et al. 1987). Recent studies indicate that the action of IL-1 involves activation of phospholipase A2 and that IL-1 has a dual effect on NGF-mRN A. Thus, IL-1 enhances NGF gene transcription and also increases the stability of NGF-mRNA in cultured sciatic nerve fibroblasts (Lindholm et al. 1988). 26 D. Lindholm et al.

The studies on the regulation of NGF synthesis in the central nervous system are less advanced compared with the peripheral nervous system. We have recently obtained evidence that IL-1 and other mesenchymal growth factors can influence NGF-mRNA levels in cultured rat astrocytes. We are currently studying the complex interplay of various growth factors in controlling NGF-mRNA synthesis in the central nervous system both in vivo and in vitro. Moreover, since it has very recently been reported that hippocampal neurons can also express NGF-mRNA (Rennert and Heinrich 1986; Ayer-LeLievre et al. 1988) this adds a new dimension to studies concerned with the regulation of NGF synthesis in the central nervous system.

References

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Hagag Ν, Halegoua S, Viola Μ (1986) Inhibition of growth factor-induced differentiation of PC 12 cells by microinjection of antibody to ras p21. Nature (London) 319:680-682 Hefti F (1986) Nerve growth factor (NGF) promotes survival of septal cholinergic neurons after fimbrial transections. J Neurosci 6:2155-2162 Hefti F, Weiner WJ (1986) Nerve growth factor and Alzheimer's disease. Ann Neurol 20:275-281 Hefti F, Dravid A, Hartikka J (1984) Chronic intraventricular injections of nerve growth factor elevate hippocampal choline acetyltransferase activity in adult rats with septo-hippocampal lesions. Brain Res 293:305-311 Heumann R, Schwab M, Thoenen Η (1981) A second messenger required for nerve growth factor biological activity? Nature (London) 292:838-840 Heumann R, Korsching S, Scott J, Thoenen Η (1984) Relationship between levels of nerve growth factor (NGF) and its messenger RNA in sympathetic ganglia and peripheral target tissues. EMBO J 3:3183-3189 Heumann R, Lindholm D, Bandtlow C, Meyer M, Radeke Μ J, Misko TP, Shooter E, Thoenen Η (1987) Differential regulation of mRNA encoding nerve growth factor and its receptor in rat sciatic nerve during development, degeneration and regeneration: role of macrophages. Proc Natl Acad Sei USA 84:8735-8739 Honegger P. Lenoir D (1982) Nerve growth factor (NGF) stimulation of cholinergic telencephalic neurons in aggregating cell cultures. Dev Brain Res 3:229-239 Hosang M, Shooter EM (1985) Molecular characteristics of nerve growth factor receptors on PC 12 cells. J Biol Chem 260:655-662 Johnson D, Larahan A, Buck CR, Sehgal A, Morgan C, Mercer E, Bothwell M, Chao Μ (1986) Expression and structure of the human NGF receptor. Cell 47:545-554 Johnson EM Jr, Rich KM, Yip Η Κ (1986) The role of NGF in sensory neurons in vivo. Trends Neurosci 1:33-37 Konkol RJ, Mailman RB, Bendeich EG, Garrison AM, Mueller RA, Breese GR (1978) Evaluation of the effects of nerve growth factor and anti-nerve growth factor on the development of central catecholaminergic neurons. Brain Res 144:277-285 Korsching S (1986) Nerve growth factor in the central nervous system. Trends Neurosci 11 /12:570-573 Korsching S, Thoenen Η (1983) Nerve growth factor in sympathetic ganglia and corresponding target organs of the rat: correlation with density of sympathetic innervation. Proc Natl Acad Sei USA 80:3513-3516 Korsching S, Auburger G, Heumann R, Scott J, Thoenen Η (1985) Levels of nerve growth factor and its mRNA in the central nervous system of the rat correlate with cholinergic innervation. EMBO J 4:1389-1393 Kromer LF (1987) Nerve growth factor treatment after brain injury prevents neuronal death. Science 235:214-216 Levi-Montalcini R (1987) The nerve growth factor: thirty-five years later. EMBO J 6:1145-1154 Levi-Montalcini R, Angeletti PU (1968) Nerve growth factor. Physiol Rev 48:534-569 Lindholm D, Heumann R, Meyer M, Thoenen Η (1987) lnterleukin-1 regulates synthesis of nerve growth factor in non-neuronal cells of rat sciatic nerve. Nature (London) 330:658-659 Lindholm D, Heumann R, Hengerer B, Thoenen Η (1988) J Biol Chem 263:16348-16351 Milbrandt J (1987) A nerve growth factor-induced gene encodes a possible transcriptional regulatory factor. Nature (London) 238:797-799 Otten U (1984) Nerve growth factor and the peptidergic sensory neurons. Trends Pharmacol 7:307-310 Pearson RCA, Sofroniew MV, Cuello AC, Powell TPS, Eckenstein F, Esiri MM, Wilcock GK (1983) Persistence of cholinergic neurons in the basal, nuclei in a brain with senile dementia of the Alzheimer's type demonstrated by immunohistochemical staining for choline acetyltransferase. Brain Res 289:375-379 Radeke MJ, Misko TP, Hsu C, Herzenberg LA, Shooter EM (1987) Gene transfer and molecular cloning of the rat nerve growth factor receptor: a new class of receptors. Nature (London) 325:593-597 Raivich G, Kreutzberg GW (1987) The localization and distribution of high affinity ßNGF binding sites in the central nervous system of the adult rat. A light microscopic autoradiographic study using (1251) ßNGF. Neuroscience 20:23-36 Rennen PD, Heinrich G (1986) Nerve growth factor mRNA inbrain: localization by in situ hy• bridization. Biochem Biophys Res Commun 138:813-818 28 D. Lindholm et al.: Nerve Growth Factor

Riopelle RJ, Verge VMK, Richardson PM (1987) Properties of receptors for nerve growth factor in the mature rat nervous system. Mol Brain Res 3:45-53 Ross AH, Grob Ρ, Bothwell Μ, Elder DE, Ernst CS, Marano Ν, Ghrist BFD, Slemp CC, Herlyn M, Atkinson Β, Koprowski Η (1984) Characterization of nerve growth factor receptor in neural crest tumours using monoclonal antibodies. Proc Natl Acad Sei USA 81:6681-6685 Schwab M, Otten U, Agid Y, Thoenen Η (1979) Nerve growth factor (NGF) in the rat CNS: absence of specific retrograde axonal transport and tyrosine hydroxylase induction in locus coeruleus and substantia nigra. Brain Res 168:473-483 Schwab ME, Heumann R, Thoenen Η (1982) Communication between target organs and nerve cells: retrograde axonal transport and site of action of nerve growth factor. Cold Spring Harbor Symp Quant Biol 46:125-134 Seiler Μ, Schwab ME (1984) Specific retrograde transport of nerve growth factor (NGF) from neocortex to nucleus basalis in the rat. Brain Res 300:33-39 Shelton DL, Reichardt LF (1984) Expression of the nerve growth factor gene correlates with the density of sympathetic innervation in effector organs. Proc Natl Acad Sei USA 81:7951-7955 Shelton DL, Reichardt LF (1986) Studies on the expression of the β nerve growth factor (NGF) gene in the central nervous system: level and regional distribution of NGF mRNA suggest that NGF functions as a trophic factor for several distinct populations of neurons. Proc Natl Acad Sei USA 83:2714-2718 Stacey DW, Watson T, Kung H-F, Curran Τ (1987) Microinjection of transforming ras protein induces c-fos expression. Mol Cell Biol 7:523-527 Sutter A, Riopelle RJ, Harris-Warrick RM, Shooter EM (1979) Nerve growth factor receptors: characterization of two distinct classes of binding sites on chick embryo sensory cells. J Biol Chem 254:5972-5982 Thoenen H, Barde Y-A (1980) Physiology of nerve growth factor. Physiol Rev 60:1284-1335 Thoenen H, Bandtlow C. Heumann R (1987) The physiological function of nerve growth factor in the central nervous system: comparison with the periphery. Rev Physiol Biochem Pharmacol 109:145-178 Thoenen H, Bandtlow C, Heumann R, Lindholm D, Meyer M, Rohrer Η (1988) Nerve growth factor: cellular localization and regulation of synthesis. Cell Mol Neurobiol 8:35-40 Whittemore SR, Ebendal T, Lärkfors L, Olson L, Seiger Α, Strömberg I, Persson Η (1986) Develop­ mental and regional expression of β nerve growth factor messenger RNA and protein in the rat central nervous system. Proc Natl Acad Sei USA 83:817-821 Will B, Hefti F (1985) Behavioural and neurochemical effects of chronic intraventricular injections of nerve growth factor in adult rats with fimbria lesions. Behav Brain Res 17:17-24 Williams LR, Varon S, Peterson GM, Wictorin K, Fischer W, Björklund A, Gage FH (1986) Continuous infusion of nerve growth factor prevents basal forebrain neuronal death after fimbria fornix transection. Proc Natl Acad Sei USA 83:9231-9235 Subject Index

Abelson murine leukemia virus (AMuLV) 38 α-tocopherylquinone 91 Acetylcholine 71,73 ft-1 peptide 113 Acute lymphoblastic leukemia (ALL) 39, 41 sequence homology to voltage-dependent Acyl-CoA oxidase deficiency 411 Na channels 114 Adducin 126 substrate for protein kinase C 118 Adiposity 257 Adrenoleukodystrophy (NALD) 420 Bcr-abl 38 Alkoxyl radicals 336 tyrosine kinase 40 Aminoacridines 90 Biliary diseases 299 3,4-aminopyridines 104 Bombesin 29,31,42-45 Amylase 73 growth factor 41 Ankyrin-band 3 interaction 65 in growth control 42 Anoxia in small-cell lung carcinomas changes in extracellular ion concentrations mitogenic 42 137 trigger for tyrosine kinase 42 suddenly occurring 137 Bradykinin 30,32 terminal depolarization 137 Brown adipose tissue 257-264, 265-284 Antibodies acute regulation 270 antimitochondrial (ΑΜΑ) 323-334 biogenesis 275 reactivity with bacteria 328 deiodinase activity 270 phosphotyrosine 37 hyperfunction 277 Antioxidants 335-343 in diabetic obese (db/db) mice 276 Antitumor drugs 359-367 in genetic obesity 276 Apamin receptor 102 in heat production 266 expression during brain ontogenesis 102 in myopathic hamsters 269 sensitive channels 102 in nude mouse 278 Arachidonic acid 286, 289 in pathological states 276 Arylsulfatase 379 in the febrile response 277 ATPase (Ca~+) activation by calmodulin 76 ligands of uncoupling protein 271 ATPase malfunction 277 (Ca2+, Mg2+) activated 71 mitochondria 259 activated by Ca2+ 75 morphological data ATPsynthase 347-358 perinatal development 266, 272 and neoplasia 350 regulation 266,270 during liver regeneration 351 thermogenetic function 257 Au-S-malate 90 ß-carotene 338 Auranofin 90 ß-cell 104 inhibition of neutrophil NADPH oxidase 90 ß-D-galactosidase 371 Autocriny 35,41,44 ß-hexosaminidase A 380 Autoerythrocyte sensitization syndrome 53 Autosensitization C-ablgene 39 against patient red blood cells 53 C-abl protein 39 antibodies against mitochondria (ΑΜΑ) 323 structural alteration 40 circulating antibodies against Ca channels 120 C-erb-B2 46,47 pathogenesis 53 C-fms 47 PS as a provocative factor 53 C-fos 24 α-tocopherol see Vitamin Ε C-kit 47 430 Subject Index

C-met 47 CPEO 200,206 C-onc proteins 47 Creatine kinase (CK) 186 C-trk 46.47 CSF-1 15 Ca2+ CSF-l-R 14 activated photoproteins 72 Cystinosis 403 activation of K+ channel 75, 99 Cytochrome c 298 free cytoplasmic concentration 77 Cytochrome c oxidase 218,228,240,260.297,360 GTP induced Ca2+ release 78 deficiencies 216-226, 228-236, 239-250 indicators 72 clinical heterogeneity 244 influx inhibited by antimycin A 74 encephalomyopathies 245 inhibition of cell entry 74 in CPEO 245 ionophores 72 in human 239 iontophoresis 72 infantile myopathies 244 mechanism of cell entry 74 lack of specific subunit 248 movements in pancreatic acinar cells 73 ophthalmoplegia 245 release type-A 246 induced by Ca2+ 78 type-B 247

induced by Ins(l,4,5)P3 78 differences in the small subunits 243 occurs in discrete organelles 78 different gene families 221 stimulated by GTP 78 from rat brown fat tissue 220 store 77 immunocytochemical studies 207 Ca2+ channels immunopurified from muscle biopsies 246 antagonists isoenzymes 243 competitive (AP7) 143 microphotometric assay 207 non-competitive (MK801) 143 nuclear and mitochondrial genome of the dihydropyridine type 143 products 228 antibodies and autoimmune disease 120 subunits 240 effectors 113 from adult and fetal tissues 219 in excitable and nonexcitable cells 112 specific antisera 224 L-type tissue-specific isoenzymes 241 in disease 119 increased in cardiomyopathic hamsters 119 D-2-amino-phosphonoheptanoate regulation by phosphorylation 117 D-2-amino-phosphonovalerate (AP5) voltage-dependent 112, 141 DAG (see also diacylglycerol) 85 purification and characterization 113 activation of protein kinase C 87 Calcineurin 117 Dantrolene 317 Calciosomes 78 DCCD-binding protein 347 Calsequestrin 78 Deiodinase 268 Carcinoma cells 30 catecholamine-induced activation 269 Cerebral metabolic rate 134 type I and II 268 Cerebro-hepato-renal (Zellweger) Dendrotoxins 108 syndrome 420 Diacylglycerol (DAG) 30, 58, 61, 73 Cholecystokinin 71 1,4-dihydropyridine (DHP) 113 Chronic myelogenous leukemia (CML) 38 DPH-as fluorescent probe 306 Chronic progressive external ophthalmoplegia Duchenne dystrophy 194 (CPEO) ^224 Cinnarizine 74 EGF (epidermal growth factor) 16. 29. 38. CI7HCO-3 antiporter 145 44-46 CP-channels chemical crosslinking 16 control by membrane potential 72 intermolecular activation 16 in the apical membrane 72 microaggregation 16 Class 1 oncogenes 36, 37 monomenc and dimeric forms 16 intracellular targets 37 signal transduction 17 Class 2 oncogenes 37 Electron transfer 177 CML 39 Elliptocytosis (herediatary) 66 Cold adaptation 265 Encephalomyelopathy 161 Complementation analysis 420 Energy metabolism 135 Complex IV 216 progressive reduction 136 Subject Index 431

Epidermal growth factor (see EGF) 29 response 23 Epilepsy with ragged red fibers (see also retrograde axonal transport 23 MERRF) 159 trophic role 23 Erb-B 14 PDGF 14,29,37,38,44,46 oncogene 29 Exocytosis 72 H-9 88 Heinz bodies 67 Fabry disease 388 Hemichromes 67 Fatal infantile mitochondrial myopathy 161, Hemolytic anemia 229 effect of hemoglobin denaturation 67 Fluorescence polarization 307 microangiopathy 66 F-met-leu-phe 89 spectrin mutants 66 Free oxygen radicals 300 transfusion-dependent 66 Hemopoietic malignancies 38 Galactosialidosis 372,373 Hereditary spherocytosis 57 Galanin 105 Histamine 30,32 Ganglioside GM3 390 human HeLa cells growth 31 Gastric carcinoma 44 Human fibroblasts 44 Gaucher disease 387 Hydrogen peroxide 336 Genetically dysgenic mouse mdg/mdg 119 Hydroxyl radicals 337 lack of receptor for Ca2+ channel effectors Hypermetabolism of non-thyroid origin 167 119 Hyperphagia 258 Glibenclamide 104 Hypertension 124-128, 129-132 Glutathione 340 defects in Na+ transport 130 Glutathione peroxidase 341 essential Glycolipid catabolism 382 animal model 124 Glycosphingolipids 379 Na+, K+ ATPase activity 131 Gold-type antirheumatic drugs 90 pregnancy-induced (PIH) 130

GM2 activator 380, 382 Hypoglycemia 146 G-proteins 32 coma 134 Growth factors 14 ionic gates involved 140 bombesin 41 preischemic 139 EGF 16 chemical crosslinking 16 Inflammation and cancer 337 intermolecular activation 16 Inositol phospholipids 29, 30

microaggregation 16 1,4,5-inositol triphosphate (seealso IP:,) 58 monomeric and dimeric forms 16 Insulin 3-13,44,45 signal transduction 17 antibodies 3 hemopoietic growth factor CSF-1 35 binding and kinase activation 4 NGF 22-28 dose-response 4 and cholinergic neurons 23 metabolic and growth-promoting effects 10 antibodies 24 phosphorylating cellular substrates 5 as a neurotrophic factor 22 receptor 17 in Alzheimer's disease 25 antiphosphotyrosine antibodies 6 insufficient supply 25 autophosphorylation 6 intracellular microinjection 24 chimeric constructions 17 levels of choline acetyltransferase domains 17 (ChAT) 23 heterodimers 17 macrophages role in regulation 25 kinase hyperactivity 6 mechanism(s) of action 24 kinase impairment 6 nature of the second messenger(s) 22 ligand-induced kinase activation 17 pharmacological modification 22 mutation 7 physiological function 22 phosphorylation 3 preventing cholinergic cell phosphorylation reversibility 6 degeneration 25 seryl (and threonly) phosphorylation 8 regional distribution 22 specificity of effects 6 regulation of synthesis 23 structure 3 required for differentiation 23 tyrosine kinase 4 432 Subject Index

release 104 phospholipase A 309

tolbutamide and sulfonylurea 104 phospholipase A2 activity 314 sensitive protein kinase 3 uncoupling of oxidative phosphorylation transmembrane signalling 3 310 Interleukin-1 25 Malignant transformation 14 Ionophore (A 23187) 58 Μ antigens 324

and phophatidylinositol 4-phosphate 59 M2 antigen 328

IP3 (Inositol-l,4,5-trisphosphate) 30, 61, 75 identity with 70 kD antigen 329 77, 85 non-mitochondrial location 329

IP4 (Inositol-l,3,4,5-tetraphosphate) 75 Mast cell degranulating peptide 107 McArdle's syndrome 192 K+ channels 57, 99-110, 105 Μ EL AS (mitonchondrial encephalomyophaty ATP-regulated 104 with lactic acidosis) 159, 223 opened by galanin 105 in complex I deficiency 159 presence ain cardiac cells 105 Membrane fluidity 285-302, 303-308 presence in neurons 105 MERFF syndrome 161, 202, 223 sulfonylureas block 104 transmission by maternal inheritance 159 Ca2+-ativated 99 Merocyanine-540 54 TEA (tetraethylammonium) block 99 binding in homologous RBC 54 voltage-dependent 107 Metachromatic leukodystrophy (MLD) 388 Kearns-Sayre syndrome (KSS) 159, 200, 206 Microcytoma 41 223 Milan hypertensive rat strain (MHS) 124 age of onset of clincal symptoms 201 Mitochondria with lacticacidemia (Robinson) 229 abnormal 189 Ketamine 149 assays of preparations 170 Kidney cross-transplantation 125 content of respiratory chain carriers 170 KSS see Kearns-Sayre syndrome diseases 157 enzyme defects Lambert-Eaton myasthenic syndrome 120 benign infantile myopathy 161 Leigh's disease (subacute necrotizing complex I 160 encephalomyelopathy) 157, 161,200 complex II 160 Liver regeneration 350 complex IV 161 Long-term potentiation 108 CPT deficiency 159 Loose coupling 157 cytochrome c oxidase (COX) 161 Luft disease 158 cytochrome c oxidase deficiency 167 Lung cancer different genetic errors 161 incidence 338 fatal infantile myopathy 161 Lysosomes 381,395-406 Krebs cycle 160 carriers for neutral amino acids 399 Leigh syndrome 161 entry of dipeptides 399 MERRF syndrome 161 hydrolases 381 oxidation-phosphorylation coupling 160 inborn errors 402 respiratory chain 160 membrane permeability 395 spontaneous mutation of mtDNA 161 metabolite-porters 399 substrate utilization 159 passive diffusion 398 fragment preparation 168 storage disease 381 in cancer chemotherapy 365 Lysosphingolipids 90 lipid composition 286 membranes amino acid composition 290 Magnetic resonance spectroscopy (see also NMR fluidity probed by spin labeling 291 and 3,P-NMR) 183 protein-lipid interactions 291 Malignant hyperthermia (MH) 309-322 morphological abnormalities 201 Malignant hyperthermia myopathies 167-180, 183-189, 200-214, 223 ATPase activity 313 abnormal morphological features 228 Ca2+ accumulation 310 cytochrome measurement 234 Ca2+ release 316 defect Ca2+ transport 310 characterization 232 halothane 310 cytochrome c oxidase 231 in healthy pigs 309 fibroblasts as diagnostic tools 230 Subject Index 433

immunodetection 232 NGF (nerve growth factor) 25 lactate to pyruvate ratio (L/P) 231 and cholinergic neurons 23 lactic acidosis 229 antibodies 24 maternal inheritance 239 in Alzheimer's disease 25 maternal transmission 212 insufficient supply 25 northern analysis 235 intracellular microinjection 24 subunit specific antisera 230 level of choline acetyltransferase (ChAT) 23 with encephalopathy 159 macrophages role in regulation 25 partial reactions of the respiratory chain 170 mechanism(s) of action 24 preparations 168 nature of the second messenger(s) 22 from Kearn-Sayre syndrome tissues 171 pharmacological modification 22 semimicro-techniques 178 physiological function 22 respiratory functions 310 preventing cholinergic cell degeneration 25 tightly coupled, from skeletal muscle 171 regional distribution 22 transformation into thermogenic 275 response 23 Mitochondrial encephalomyopathy with lactic Nifedipine 74 acidosis (see also MELAS) 159 NMR 183 Morris hepatoma 3924A 351 ATP concentration 184 Muscular dystrophy 194 determination of intracellular pH 183 Myopathies isolated heart mitochondria in vitro 185 megaconial 157 phosphocreatine (PCr) and inorganic mitochondria 187.200 phosphates (Pj) detection 183 carnitine palmitoyltransferase (CPT) spectroscopy 158 deficiency 157 enzyme defects Obesity 257 complex I 200 genetic forms complex III 200 db/db mouse 258 complex IV 200 ob/ob mouse 258 major gene deletions 158 synthesis and turnover of cholesterol 303 microphotometric assays 200 Omega-phosphonic acid 143 morphological classification criteria 157 Oncogenes (see also specific oncogenes) NADH-coenzyme Q reductase 188 class -2 35 MP nuclear magnetic resonance (NMR) coded tyrosine kinases 36 spectroscopy 158 control of cell proliferation 36 ragged red fibers. RRF 157 tyrosine phosphorylation 36 reduction in post-excrcise recovery of Organic hydroperoxides 336 PCr 187 Oxidative phosphorylation 134, 216, 359 steroid experimental therapy 195 in rapidly growing tumors

vitamins K:> (menadione) therapy 194 respiratory chain-linked 176 pleoconial 157 uncoupling 266 Myotonic muscle dystrophy 102 Oxytetracycline 362

N-acetyl-a-neuraminidase (sialidase) 371 P|45 44 Na+ channel 99 biological functions 45 sensitivity to apamin 99 dephosphorylation 46 Na+ retention in MHS 125 in liver cells 46 Na+/Ca2+exchange 71,76 tyrosine phosphorylation 45 Na+/H+ antiporter 145 Pancreatic acinar cell 71 Na+/H+ exchange PCr/Pj 190 activation of 32 age-related reduction 194 amiloride-sensitive 32 at rest 193 deactivation 33 carnitine deficiency 190 differentiation process 33 in acute disorders 195 NADH-dehydrogenase (complex I) 25 in chronic polymyositis 194 Neoplastic cell markers 37 in resting muscle PCr/Pj 195 Neu , 15 malignant hyperthermia 190 Neuroblastoma 89 non specificity for primary mitochondrial Neurohormone 42 diseases 196 434 Subject Index

recovery by unsaturated fatty acids 86 after exercise 193 and EGF receptor phosphorylation 86 in myophosphorylase deficiency 192 apparent molecular weight 84 PDGF 37,38,44,46 auto-phosphorylation of α and γ forms 86 Peroxisomes 407 differential distribution in brain 85 /3-oxidation 407-419 in human neutrophils 85 energetic aspects 410 in normal and ras-transformed 3T3 inborn errors 411 fibroblasts 85 induction 410 inhibitors 88,90 in detoxification reactions, serving 415 affecting Ca2+ binding 88 inherited disorders 420-428 affecting the site of interaction with the in mammalian metabolism 407 protein substrate 88 physiological importance 407 platelet aggregation 89 thiolase deficiency 411 redistribution 85 visualization by indirect immuno­ regulatory domain 86

fluorescence 423 preparation by tryptic digestion 86 v Phenylalkylamines (PAA) 113 substances affecting 89 Philadelphia chromosome 38 respiratory burst of neutrophils 89 Phorbol esters 33,73 Protein transport 364 Phosphatidylethanolamine (PE) 53 Proton conductivity 353 Phosphatidylinositol 4,5-biphosphate Pseudogenes 223

(PIP2) 58 Pyropoikilocytosis 66 Phosphatidylserine (PS) 53 Phosphatidylserine reaction with TNBS 54 Quin 2 72 Phosphoinositide signalling system 58 Phospholipase C 29, 73 Ragged-red fibers 201,223 Phospholipids Ras 24 asymmetry 53,54 Ras-like proteins 24 plasma membrane 53 RBC (red blood cells) reaction of RBC with TNBS 55 cytoskeleton 55 Phosphorylation potential (PP) 186 visualization in human cells 55 Phosphotyrosine antibodies 37 merocyanine 540 binding 55 Plasma phospholipids reaction with TNBS 55 abnormalities in hypertension 124-128 presence of tubulin and actin 55 lipids 304 tubulin in human cells 55 thermolabile factor 56 Receptors 31P-NMR 183-198 aggregation 18 indicators of mitochondrial disease 189 and channel regulation 112 monitoring of therapy 194 autoregulation 37 spectrum of muscle at rest 183 bombesin complex in small-cell lung Primary biliary cirrhosis (PBC) 323-334 carcinomas 43 antimitochondrial antibodies (ΑΜΑ) 323 cellular activation 18 cross-reacting epitopes 332 collision-coupling 18 oxygen-free radicals (oxidants) 335 dimerization 18 pathogenesis 332 EGF 16 Progressive external ophthalmoplegia for apamin 102 (see also CPEO) 200 G-protein coupled 15 Protective protein 371-378 gl uta mate 142 cDNA 373 activation Protein kinase C 30, 38, 58, 73, 84-94,127 kainate (K) 142 α,β and γ forms 85 N-methyl-D-aspartate (N or activation of 31, 84, 85 NMDA) 142 by aplysiatoxin 87 quisqualate (Q) 142 by DAG growth factors 29-34 by lead 87 lacking tyrosine kinase activity 24 by phorbol diesters 87 monoclonal and polyclonal antibodies 18 by teleocidin 87 PDGF 45 by tumor promoters 87 photoaffinity labelling 113 Subject Index 435

signal transduction by EGF 16 Thermogenesis 265 tyrosine kinase 14-21, 32, 35-52 mechanism 267 tyrosine kinases (RTK's) 14 Thermoregulation 265 Red blood cell (see also RBC) 53 Thrombin 31 membrane skeleton 64 Thyroid hormone 268 stability and oxidant damage 67 TLCK (N-tosyl-L-lysine chloromethvl ketone) Renal transplant rejection 67 88 Respiratory burst of neutrophils 89 Trifluoperazine 88 Respiratory chain 239 Trolox 91 abnormalities 210 Tumor promotion 89 tissue-specific isoforms of enzymes 212 activation of protein kinase C 84 Respiratory control ratio (RCR) 353 Tyrosine kinase 32 Retroposons 222 activated in human tumors 44 Retropseudogenes 223 Ros 15 Ubiquinone-cytochrome c oxidoreductase RTK (complex III) 216 overexpression 14 Uncoupling protein 259, 266 signal transduction 15 as dimer 267 GDP binding 274 Salla disease 400 latent form 272 Sandhoff disease 383 long-term regulation 272 Secretion of digestive enzymes masking-unmasking 272 2+ intracellular Ca as a regulator 72 mRNA-synthesis 275 Sickle cells 57 Unsaturated fatty acids 289 Spectrin 61,64 abnormalities of molecule 65 V-fes 46 hemolytic anemia 66 V-src 46 mutants 66 Viral oncogenes 14 Sphingosine 90 Vitamin C Staurosporine 88,89 antioxidant, anticancer, anti-aging effects N-propan-2-ol- and N-phenyl-derivatives 88 338 Steinen disease 102 Vitamin Ε 90, 338 Storage diseases 379-394 anticancer, anti-aging effects 338 Subacute necrotizing encephalomyelopathy antioxidant role 90, 338 (Leigh's disease) 229 differentiation in human neuroblastoma 90 Substance Ρ 29 H202 production inhibition in human Sulfatide activator 386 neutrophils 90 in glycolipid catabolism 387 in plasma 91 Superoxide dismutase 341 platelet anti-aggregation agent 90 Tay-Sachs disease 383 TEMPO (2,2,6,6, tetramethylpiperidine-l-oxyl) 292 X-linked adrenodystrophy 411 fluidity of human membranes 293 Zellweger syndrome 411,420