Handbook of

Experimental Pharmacology
Volume 1041/
Editorial Board
G.V.R. Born, London
P. Cuatrecasas, Ann Arbor, MI
H. Herken. Berlin
Opioids I
Contributors
H. Akil, S. Archer, A. Beaumont, M. Blum, D. Bronstein, S.R. Childers
A.D. Corbett, B.M. Cox, V. Dauge, R. Day, S. Dermer, J. Donnerer
A.W. Duggan, R. Elde, S.M. Fleetwood-Walker, L.D. Fricker
M.-C. Fournie-Zaluski, T.L. Gioannini, A. Goldstein, T. H6kfelt
V. H6llt, J.W. Holaday, H. Khachaturian, N. Kley, C.M. Knapp
H.W. Kosteriitz, N.M. Lee, F.M. Leslie, N. Levin, M.E. Lewis
J.P. Loeffler, H.H. Loh, D. Lorang, J.R. Lundblad, A. Mansour
A.H. Mulder, R.A. North, S.J. Paterson, D.E. Pellegrini-Giampietro
J.E. Pintar, F. Porreca, P.S. Portoghese, J.L. Roberts, B.R. Roques
J. Rossier, R.B. Rothman, M.K.H. Schafer, P.W. Schiller
A.N.M. Schoffelmeer, R.E.M. Scott, E.J. Simon, A.P. Smith
J.A.M. Smith, S. Spector, A. Tempel, H. Teschemacher, K.A. Trujillo
E. Young, S.J. Watson, P.L. Wood, R.S. Zukin

Editor: Albert Herz

Section Editors: H. Akil and E.1. Simon

Springer-Verlag
Berlin Heidelberg New York London Paris
Tokyo Hong Kong Barcelona Budapest
Professor Dr.med. ALBERT HERZ
Max-Planck-Institut fur Psychiatrie
Abteilung Neuropharmakologie
Am Klopferspitz 18
W-8033 Martinsried, FRG

Section Editors
Dr. med. HUDA AKIL
Mental Health Research Institute
School of Medicine
University of Michigan
205 Zina Pitcher
Ann Arbor, MI 48109, USA
Dr. med. ERIC J. SIMON
New York University
Medical Center
School of Medicine
550 First A venue
New York, NY 10016, USA

With 74 Figures and 52 Tables

ISBN -13: 978-3-642-77462-1 e- ISBN -13: 978-3-642-77460-7

DOl: 10.1007/978-3-642-77460-7

Library of Congress Cataloging-in-Publication Data. Opioids I / contributors, H. Akil ... [et al.l;
editor, Albert Herz; (H. Akil and EJ. Simon, section editors). p. cm. - (Handbook of experimental
pharmacology; v. 104) Includes bibliographical references and index.
ISBN-13:978-3-{i42-77462-1 l. Opioids. 2. Opioids - Receptors. I. Akil, H.
(Huda) II. Herz, Albert, 1921- . III. Simon, Eric 1. IV. Title: Opioids l. V. Series. [DNLM:
1. Endorphins. 2. Receptors, Endorphin. WI HA51L v. 104 / QU 68 0603] QP905.H3 vol. 104
[RM328] 615' .1 s - dc20 [615' .78] DNLMIDLC for Library of Congress 92-2325 CIP
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List of Contributors

AKIL, H., Mental Health Research Institute, School of Medicine, University

of Michigan, 205 Zina Pitcher, Ann Arbor, MI 48109, USA
ARCHER, S., Department of Chemistry, Cogswell Laboratory, Rensselaer
Polytechnic Institute, Troy, NY 12180-3590, USA
BEAUMONT, A., Departement de Chimie Organique, U 266 INSERM,
UA 498 CNRS, UFR des Sciences Pharmaceutiques et Biologiques,
4, avenue de l'Observatoire, F-75006 Paris, France
BLUM, M., The Fishberg Research Center in Neurobiology, Mount Sinai
School of Medicine, One Gustave L. Levy Place, New York, NY 10029,
USA
BRONSTEIN, D., Laboratory of Molecular Integrative Neuroscience, National
Institute of Environmental Health Sciences, Research Triangle, Park,
NC 27709, USA
CHILDERS, S.R., Department of Physiology and Pharmacology, Bowman
Gray School of Medicine, Medical Center Boulevard, Winston-Salem,
NC 27157-1083, USA
CORBETT, A.D., Department of Biomedical Sciences, University of
Aberdeen, Marischal College, Aberdeen AB9 lAS, Great Britain
Cox, B.M., Department of Pharmacology, Uniformed Services University
of the Health Sciences, 4301 lones Bridge Road, Bethesda, MD 20814-
4799, USA
DAUGE, V., Departement de Chimie Organique, U 266 INSERM, UA 498
CNRS, UFR des Sciences Pharmaceutiques et Biologiques, 4, avenue
de l'Observatoire, F-75006 Paris, France
DAY, R., I.A. DeSeve Laboratory of Biochemical and Molecular Neuro-
endocrinology, Clinical Research Institute of Montreal, 110 Pine
Avenue West, Montreal, Quebec, Canada H2W lR7
DERMER, S., The Fishberg Research Center in Neurobiology, Mount Sinai
School of Medicine, One Gustave L. Levy Place, New York, NY 10029,
USA
VI List of Contributors

DONNERER, J., Department of Experimental and Clinical Pharmacology,

University of Graz, A-801O Graz, Austria
DUGGAN, A.W., Department of Preclinical Veterinary Sciences, Royal
(Dick) School of Veterinary Studies, University of Edinburgh,
Summerhall, Edinburgh EH9 1QH, Great Britain
ELDE, R., University of Minnesota, Department of Cell Biology and
Neuroanatomy, 321 Church Street SE, Minneapolis, MN 55455, USA
FLEETWOOD-WALKER, S.M., Department of Preclinical Veterinary Sciences,
Royal (Dick) School of Veterinary Studies, University of Edinburgh,
Summerhall, Edinburgh EH9 1QH, Great Britain
FRICKER, L.D., Department of Molecular Pharmacology, Albert Einstein
College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461,
USA
FOURNIE-ZALUSKI, M.-C., Departement de Chimie Organique, U 266
INSERM, UA 498 CNRS, UFR des Sciences Pharmaceutiques et
Biologiques, 4, avenue de l'Observatoire, F-75006 Paris, France
GIOANNINI, T.L., Department of Psychiatry, New York University Medical
Center, 550 First Avenue, New York, NY 10016, and Baruch College,
CUNY, New York, NY 10010, USA
GOLDSTEIN, AVRAM, Stanford University, 735 Dolores, Stanford, CA 94305,
USA
HOKFELT, T., Karolinska Institute, Department of Histology and Neuro-
biology, P.O. Box 60 400, S-104 01 Stockholm, Sweden
HOLLT, V., Physiologisches Institut, Universitat Munchen, PettenkoferstraBe
12, W-8000 Munchen, FRG
HOLADAY, J.W., Medicis Corporation, 100 East 42nd Street, 15th Floor,
New York, NY 10017, USA
KHACHATURIAN, H., Molecular and Cellular Neuroscience Research Branch,
National Institute of Mental Health, Room llC-05, Parklawn Building,
Rockville, MD 20857, USA
KLEY, N., Molecular Neurooncology Laboratory, Massachusetts General
Hospital, Harvard Medical School, 149 13th Street, Charlestown, MA
02129-9142, USA
KNAPP, C.M., Department of Pharmacy, Veterans Admin. Hospital, 130 W.
Kingsbridge Road, Bronx, NY 10468, USA
KOSTERLITZ, H.W., Unit for Research on Addictive Drugs, Marischal
College, University of Aberdeen, Aberdeen AB9 lAS, Great Britain
List of Contributors VII

LEE, N.M., Department of Pharmacology, University of Minnesota Twin

Cities, Medical School, 3-249 Millard Hall, 435 Delaware Street, S.E.,
Minneapolis, MN 55455, USA
LESLIE, F.M., Department of Pharmacology, California College of
Medicine, University of California, Irvine, CA 92717, USA
LEVIN, N., The Fishberg Research Center in Neurobiology, Mount Sinai
School of Medicine, One Gustave L. Levy Place, New York, NY 10029,
USA
LEWIS, M.E., Cephalon, Inc., 145 Brandywine Parkway, West Chester, PA
19380, USA
LOEFFLER, J.P., Laboratoire de Physiologie Generale, IPCB, 21, rue Rene
Descartes, F-67100 Strasbourg Cedex, France
LOH, H.H., Department of Pharmacology, University of Minnesota Twin
Cities, Medical School, 3-249 Millard Hall, 435 Delaware Street, S.E.,
Minneapolis, MN 55455, USA
LORANG, D., The Fishberg Research Center in Neurobiology, Mount Sinai
School of Medicine, One Gustave L. Levy Place, New York, NY 10029,
USA
LUNDBLAD, J.R., The Fishberg Research Center in Neurobiology, Mount
Sinai School of Medicine, One Gustave L. Levy Place, New York, NY
10029, USA
MANSOUR, A., Mental Health Research Institute, University of Michigan,
205 Zina Pitcher Place, Ann Arbor, MI 48109-0720, USA
MULDER, A.H., Department of Pharmacology, Free University Medical
Faculty, Van der Boechorststraat 7, NL-1081 BT Amsterdam, The
Netherlands
NORTH, R.A., Vollum Institute, Oregon Health Sciences University,
3181 SW Sam Jackson Park Road, Portland, OR 97201, USA
PATERSON, S.J., U.M.D.S., Guy's and St. Thomas's Medical and Dental
School, Department of Pharmacology, Lambeth Palace Road, London
SE1 7EH, Great Britain
PELLEGRINI-GIAMPIETRO, D.E., Department of Neuroscience, Albert Einstein
College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461-1988,
USA
PINTAR, J.E., Department of Anatomy and Cell Biology, Columbia College
of Physicians and Surgeons, 630 W. 168th Street, New York, NY 10032,
USA
VIII List of Contributors

PORRECA, F., Department of Pharmacology, College of Medicine, University

of Arizona, Tucson, AZ 85724, USA
PORTOGHESE, P.S., Department of Medicinal Chemistry, College of Phar-
macy, University of Minnesota, 308 Harvard Street, S.E., Minneapolis,
MN 55455, USA
ROBERTS, J.L., The Fishberg Research Center in Neurobiology, Mount Sinai
School of Medicine, One Gustave L. Levy Place, New York, NY 10029,
USA
ROQuEs, B.R., Departement de Chimie Organique, U 266 INSERM,
UA 498 CNRS, UFR des Sciences Pharmaceutiques et Biologiques,
4, avenue de l'Observatoire, F-75006 Paris, France
ROSSlER, J., Institut Alfred Fessard, Centre National de la Recherche
Scientifique, F-91198 Gif-sur-Yvette Cedex, France
ROTHMAN, R.B., Laboratory of Clinical Psychopharmacology, NIDA
Addiction Research Center, P.O. Box 5180, Baltimore, MD 21224,
USA
SCHAFER, M.K.H., Anatomisches Institut, Johannes-Gutenberg-Universitat,
SaarstraBe 21, W-6500 Mainz, FRG
SCHILLER, P. W., Laboratory of Chemical Biology and Peptide Research,
Clinical Research Institute of Montreal, 110, avenue des Pins Ouest,
Montreal, Quebec, Canada H2W 1R7
SCHOI'l'ELMEER, A.N .M., Department of Pharmacology, Free University
Medical Faculty, Van der Boechorststraat 7, NL-1081 BT, Amsterdam,
The Netherlands
SCOTT, R.E.M., Department of Anatomy and Cell Biology, Columbia
College of Physicians and Surgeons, 630 W. 168th Street, New York,
NY 10032, USA
SIMON, E.J., Departments of Psychiatry and Pharmacology, New York
University Medical Center, 550 First Avenue, New York, NY 10016,
USA
SMITH, A.P., Department of Pharmacology, University of Minnesota Twin
Cities, Medical School, 3-249 Millard Hall, 435 Delaware Street, S.E.,
Minneapolis, MN 55455, USA
SMITH, J .A.M., Department of Pharmacology, California College of
Medicine, University of California, Irvine, CA 927l7, USA

SPECTOR, S., Vanderbilt University Medical School, Department of Psy-

chiatry and Pharmacology, Nashville, TN 37232, USA
List of Contributors IX

TEMPEL, A., Department of Psychiatry, Hillside Hospital, Division of

L.I.J.M.C., 266th Street and 16th Street Avenue, Glen Oaks, NY
11004, USA
TESCHEMACHER, H., Rudolf-Buchheim-Institut fUr Pharmakologie der
Justus-Liebig-Universitat, Frankfurter StraBe 107, W-6300 GieBen, FRG
TRUJILLO, K.A., Mental Health Research Institute, University of Michigan,
205 Zina Pitcher Place, Ann Arbor, MI 48109-0720, USA
YOUNG, E., Mental Health Research Institute, University of Michigan, 205
Zina Pitcher Place, Ann Arbor, MI 48109-0720, USA
WATSON, SJ., Mental Health Research Institute, University of Michigan,
205 Washtenaw Place, Ann Arbor, MI 48109-0720, USA
WOOD, P.L., Mayo Clinic Jacksonville, 4500 San Pablo Road, Research
Building 3, Jacksonville, FL 32224, USA
ZUKIN, R.S., Department of Neuroscience, Albert Einstein College of
Medicine, 1300 Morris Park Avenue, Bronx, NY 10461-1988, USA
HANS KOSTERLITZ
Dedication

It is a great pleasure to dedicate this volume to Professor HANS KOSTERLlTZ,

of the University of Aberdeen, Scotland. Professor KOSTERLITZ is un-
doubtedly one of the leaders in the opioid field, whose scientific vision has
exerted a significant impact on the development of this field of research. His
career represents a lifetime of successful and important research, which
culminated after his official retirement, when he and his colleague, Dr.
JOHN HUGHES, along with a dynamic scientific team, discovered the first
endogenous opioid peptides, the enkephalins.
Dr. KOSTERLITZ received his M.D. degree in 1928 from the University of
Berlin and worked in that medical school until 1933. He then decided to
leave Germany and took a position in the Physiology Department at the
University of Aberdeen, where Nobel laureate J.J.R. MACLEOD held the
chair. His research on carbohydrate metabolism bore fruit in 1937 when he
isolated galactose-I-phosphate, an important intermediate in the conversion
of galactose to glucose in the liver. In 1968, Dr. KOSTERLITZ was appointed
to the newly created chair of pharmacology. In 1973, he retired at the age of
70 and became professor emeritus. The same year he was named director of
a new laboratory established at the University of Aberdeen, called the Unit
for Research on Addictive Drugs.
Starting in the 1940s his research increasingly began to reflect his inter-
est in neuroscience and particularly in the mode of action of morphine and
related narcotic analgesics. In the early 1950s, he demonstrated that the
isolated guinea pig ileum is an excellent bioassay system for these drugs. In
spite of ridicule by scientists who felt that any work not directly concerned
with the central nervous system was irrelevant, he persisted. In extensive
and thorough research, he clearly demonstrated the usefulness of in vitro
bioassay systems for the study of opiates. His laboratory has developed
several bioassay systems in addition to the guinea pig ileum. These include
the mouse vas deferens and, more recently, the hamster and rat vas deferens
systems, all of which are now used throughout the world for the assay and
differentiation of the various types of opioid receptors.
It was also the use of the in vitro systems that permitted HANS
KOSTERLITZ and his young collaborators, most notably JOHN HUGHES, to
isolate from pig brain the first endogenous peptides with opiate-like activity
in 1975, which they named methionine- and leucine enkephalin. This was a
XII Dedication

major discovery which catalyzed major advances in the field of neuropeptide

research, an area which has since become one of the most active in neuro-
science. It is of interest that this discovery was made two years after Dr.
KOSTERLITZ' official retirement.
Professor KOSTERLITZ continued his work on opioids, focusing on the
mechanism of release of enkephalins and on the multiplicity of opioid
receptors. His wide ranging interests have encompassed almost every level
of study of endogenous opioids, from physical structure, pharmacology,
regulation, to overall physiological functions. It is, therefore, fitting that this
volume, which attempts to cover the multiple levels of discourse in the study
of endogenous opioids, be dedicated to him.
Looking back at Professor KOSTERLITZ' career, one is struck by two
remarkable characteristics: First, he has changed his areas of scientific interest
more than once, and the changes have not been minor (e.g. from carbo-
hydrate metabolism to endogenous opioids), and second, he made signi-
ficant and lasting contributions in each of these fields.
To have altered the course of research in one area is a feat and an
honor, yet to have done so repeatedly is a sign of true scientific genius.
Interestingly, Professor KOSTERLITZ attributes 1 his success to the very fact
that he changed his research interests during the course of his career, a
process which he says suited him "temperamentally." Dr. KOSTERLITZ
asserts that "such a change creates a challenge to become familiar with new
concepts and then try to compete as a newcomer." While this may crea~
"worrisome complications," he believes that the difficulties "are helpful in
postponing the inevitable losses in flexibility and adaptability."
We can think of no one who better exemplifies these characteristics than
Professor KOSTERLITZ. We hope that this volume will inspire similarly fresh
and novel ideas among its readers.
The Editors

I From H.W. KOSTERLITZ (1979) The best laid schemes 0' mice an' men gang aft
agley. Ann Rev Pharmacol Toxicol 19:1-12.
Preface

In 1957 Otto SCHAUMANN, one of the pionieers in pharmacological research

on morphine and the first to prepare synthetic opiates, presented a mono-
graph entitled "Morphin und morphiniihnliche Verbindungen" as Volume
12 of the Handbook of Experimental Pharmacology. Now, 35 years later, we
are publishing in the same series a new comprehensive volume covering the
present status of opioid research. Since that time the topic has expanded
enormously. The identification of opioid receptors and the detection of their
endogenous ligands were landmarks which opened a new era in opioid
research and fertilized the entire field of neurobiology. The rapid develop-
ment of this field is illustrated in the figure, which represents the number of
papers published on opioid research since 1970 (searches performed on the
MEDLINE data base).

3000

Vl
<-
QJ
a.
rt!
a. 2000
....
0
<-
QJ
..0
E
:::J
C
1000

1970 75 80 85 1990
year
Fig. 1. The number of papers published on opioids between 1970-1991 (Medline
data base). Key, 0 - 0 endorphins, 0 - 0 opiate/opioid receptors, f',-f', narcotic
dependence, . - . opioids (total, includes morphine, morphine derivatives,
endorphins, opiate/opioid receptors, and narcotic dependence)
XIV Preface

Attempts to bring out a similar state-of-the-art volume some 10-12

years ago had to be abandoned due to the logarithmic escalation in the
amount of opioid research being performed at that time. Highly stimulating
and often important papers on opioid research continue to appear in large
numbers, but the growth phase seems to have reached a plateau. Although
our knowledge on the molecular structure of opioid receptors is still very
preliminary, this seems to be an appropriate time to summarize the current
status of this most stimulating research domain.
Given the immense amount of literature which has accumulated on
opioids during recent years and the limited space available in this publica-
tion, we were faced with the difficult task of selecting what we considered to
be the most important topics of current and earlier research. We hope that
our judgement was fair.
Part I deals with the multiplicity of opioid receptors (characterization,
distribution, regulation etc.), the chemistry of opiates and the biochemistry
and molecular biology of opioid peptides (gene expression, biosynthesis,
inactivation, receptor selectivity of ligands, etc.), and the neurophysiology
of opioids and their moleclular actions. Part II reviews a broad spectrum
of physiological and behavioral functions and pharmacological actions of
opioids. In addition, the neuroendocrinology of opioids as well as opioid
tolerance and dependence are discussed in a series of chapters. The final
part deals with the pathophysiology and clinical uses of opioids. Some
unavoidable overlap occurs between several chapters, but the detailed
subject index should help orient readers.
We would like to express our sincere thanks to the authors, who wrote
their chapters without too much delay, enabling us to present up-to-date
accounts. We are indepted to Prof. H. HERKEN, who initially suggested the
writing of this treatise. Finally, we would like to express our gratitude to
Mrs. DORIS WALKER and the other staff at Springer-Verlag as well as to our
secretaries for their efforts in bringing this work to fruition.
Contents

Section A: Opioid Receptors/Multiplicity

CHAPTER 1
Opioid Receptor Multiplicity: Isolation, Purification,
and Chemical Characterization of Binding Sites
E.J. SIMON and T.L. GIOANNINI. With 3 Figures. . . . . . . . . . . . . . . . . . . . . 3
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
B. Opioid Receptors Exist in Multiple Types. . . . . . . . . . . . . . . . . . . . . . . 3
C. Selective Ligands for the Major Types of Opioid Receptors ....... 5
D. Characterization of Membrane-Bound Opioid Receptor Types. . . . . 6
E. Putative Endogenous Ligands . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
F. Separation and Purification of Opioid Binding Sites .............. 9
I. Solubilization . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
II. Physical Separation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 10
III. Affinity Cross-Linking ................................... 11
IV. Partial Purification. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
V. Purification to Homogeneity .............................. 15
G. Recent Studies on Purified ~-Opioid Binding Protein . . . . . . . . . . . .. 18
I. Antibodies Generated Against Peptide Sequences. . . . . . . . . . .. 18
II. Rhodopsin Antibodies React with Purified OBP . . . . . . . . . . . .. 19
III. Attempts to Clone the cDNA of Purified OBP . . . . . . . . . . . . . .. 20
H. Concluding Comments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 20
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 21

CHAPTER 2
Expression Cloning of cDNA Encoding a Putative Opioid Receptor
A. GOLDSTEIN. With 1 Figure. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 27
A. Project History. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 27
B. Expression Cloning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 28
I. Methodology ........................................... 28
II. Attempt by Stable Transfection. . . . . . . . . . . . . . . . . . . . . . . . . . .. 29
III. Transient Transfection, Panning. . . . . . . . . . . . . . . . . . . . . . . . . .. 30
XVI Contents

C. Ligand Binding by the Expressed Receptor ..................... 32

D. Sequence Analysis, Structure of the Receptor. . . . . . . . . . . . . . . . . .. 32
E. Conclusions ................................................ 35
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 35

CHAPTER 3
Characterization of Opioid-Binding Proteins
and Other Molecules Related to Opioid Function
A.P. SMITH, H.H. LOH, and N.M. LEE. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 37
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 37
B. cDNA Cloning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 37
I. Molecular Cloning of OBCAM . . . . . . . . . . . . . . . . . . . . . . . . . . .. 41
II. Molecular Cloning and Characterization of Gene Products
Downregulated by Chronic Opioid Treatment
ofNG108-15 Cells....................................... 43
III. Use of Consensus Sequences in cDNA Cloning
of Opioid Receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 45
C. Use of Antibodies to Characterize Opioid Receptors . . . . . . . . . . . .. 45
D. Antisense cDNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 48
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 49

CHAPTER 4
Use of Organ Systems for Opioid Bioassay
J.A.M. SMITH and F.M. LESLIE. With 7 Figures 53
A. Introduction................................................ 53
I. Rationale for the Use of Isolated Organ Systems. . . . . . . . . . . .. 53
II. Tissue Preparations. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 53
III. Applications of Peripheral Tissue Bioassay. . . . . . . . . . . . . . . . .. 54
B. Measurement of Pharmacological Constants. . . . . . . . . . . . . . . . . . . .. 55
I. Theoretical Considerations ... . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 55
1. Determination of Agonist Affinity ....................... 55
2. Determination of Antagonist Affinity. . . . . . . . . . . . . . . . . . . .. 57
II. Methodological Considerations. . . . . . . . . . . . . . . . . . . . . . . . . . . .. 59
1. Choise of Tissue Preparation. . . . . . . . . . . . . . . . . . . . . . . . . . .. 59
2. Tissue Preparation and Setup. . . . . . . . . . . . . . . . . . . . . . . . . . .. 59
3. Optimization of Equilibrium Conditions .... . . . . . . . . . . . . .. 60
C. Assay Preparations . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 63
I. Guinea Pig Ileum. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 63
1. Il-Receptors.......................................... 63
2. K-Receptors.......................................... 66
3. i)-Receptors.......................................... 66
Contents XVII

II. Mouse Vas Deferens. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 67

1. ~-Receptors.......................................... 67
2. K-Receptors.......................................... 68
3. (i-Receptors.......................................... 69
III. Other Vasa Deferentia. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 69
1. Rat Vas Deferens. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 69
2. Hamster Vas Deferens ................................ 72
3. Rabbit Vas Deferens. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 72
D. Conclusions ................................................ 73
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 74

CHAPTER 5
Anatomical Distribution of Opioid Receptors in Mammalians:
An Overview
A. MANSOUR and S.l. WATSON. With 6 Figures. . .. . . . .. . ... . . . . .. . .. 79
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 79
B. Anatomical Distributions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 81
I. ~-Receptors ............................................ 81
II. o-Receptors ............................................ 90
III. K-Receptors ............................................ 92
IV. Anatomical Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 93
C. Multiple K-Receptor Subty'pes. . .. . . .. . . .. . . .. . . . .. . ... .. . . . . .. 94
D. Nigrostriatal and Mesolimbic Dopamine Systems as Models
for Opioid Peptide and Receptor Interactions ................... 97
I. Conclusions............................................. 100
E. Future Directions ........................................... 101
References ..................................................... 102

CHAPTER 6
Opioid Receptor Regulation
R.S. ZUKIN, D.E. PELLEGRINI-GIAMPIETRO, C.M. KNAPP,
and A. TEMPEL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 107
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 107
B. Regulation of Opioid Receptors in the Adult Brain
by Chronically Administered Opioid Agonists and Antagonists .... 110
I. Chronic Administration of Opioid Agonists In Vivo . . . . . . . . .. 110
II. Chronic Administration of Agonists to Cells Grown in Culture 111
III. Chronic Administration of Opioid Antagonists .............. 112
C. Regulation of Opioid Receptors by Other Drugs
or Specific Brain Lesions ..................................... 114
D. Regulation of Opioid Receptor and Peptide Gene Expression
in Embryonic and Neonatal Brain. . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 115
XVIII Contents

I. Effects of Chronic Opioid Administration

on Opioid Receptor Expression ............................ 116
1. Perinatal Treatment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 116
2. Postnatal Treatment ................................... 117
II. Effects of Chronic Opioid Administration
on Opioid Peptide Expression. . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 118
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 119

CHAPTER 7
Multiple Opioid Receptors and Presynaptic Modulation
of Neurotransmitter Release in the Brain
A.H. MULDER and A.N.M. SCHOFFELMEER. With 4 Figures 125
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 125
B. Modulation of Noradrenaline Release. . . . . . . . . . . . . . . . . . . . . . . . .. 127
C. Modulation of Acetylcholine Release. . . . . . . . . . . . . . . . . . . . . . . . . .. 130
D. Modulation of Dopamine Release ............................. 134
E. Modulation of the Release of Other Neurotransmitters ..... . . . . .. 137
F. Conclusions ................................................ 138
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 140

CHAPTER 8
Opioid Receptor-G Protein Interactions:
Acute and Chronic Effects of Opioids
B.M. Cox. With 2 Figures ....................................... 145
A. Introduction................................................ 145
B. Effects of Guanine Nucleotides on Ligand Binding
to Opioid Receptors ......................................... 147
I. Opioid 11- and b-Receptors Are Funtionally Linked
to Guanine Nucleotide Binding Proteins. . . . . . . . . . . . . . . . . . .. 147
1. Guanine Nucleotides Lower Agnonist Affinity
at 11- and b-Receptors .................................. 147
2. Guanine Nucleotides Increase Agonist Dissociation Rates.. 148
3. Guanine Nucleotide Effects on Equilibrium Binding
of Opioids ........................................... 149
4. Sodium Regulates Agonist Affinity at 11- and b-Receptors. .. 156
5. Stimulation of GTPase Activity by Activation
of 11- and b-Receptors ................................. 157
II. Evidence for K-Receptor Interactions with G Proteins ........ 158
1. Effects of Guanine Nucleotides on Agonist Binding
at K,-Sites. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 159
2. Effects of Guanine Nucleotides on Binding at Kz-Sites. . . . .. 159
III. Stimulatory Effects of Opioids:
Possible Interactions of Opioid Receptors with G s . . . . . . . . . . .. 160
Contents XIX

C. Cellular Consequences of Sustained Exposure to Opiate Drugs 162

I. Characteristics of Opioid Tolerance and Dependence. . . . . . . .. 162
II. Changes in the Number of Opioid Receptors Following
Sustained Exposure to High Concentrations of Opiate Drugs .. 164
1. In Vitro Studies Employing Tissue Culture . . . . . . . . . . . . . .. 164
2. Effects of Chronic Opioid Treatment in Brain. . . . . . . . . . . .. 165
3. Effects of Chronic Treatment with K-Agonists. . . . . . . . . . . .. 167
4. Mechanisms Implicated in Changes
in Receptor Site Density. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 168
III. Chronic Opioid Treatment Uncouples Opioid Receptors
from Their Associated G Proteins. . . . . . . . . . . . . . . . . . . . . . . . .. 169
1. Receptor Desensitization; 11- and b-Receptors. . . . . . . . . . . .. 169
2. Mechanisms Implicated in Receptor Desensitization . . . . . .. 170
IV. Sustained Opioid Exposure Induces Changes
in the Cellular Concentrations of Some G Proteins . . . . . . . . . .. 172
1. Neuroblastoma X Glioma (NG 108-15) Hybrid Cells. . . . . .. 172
2. Guinea Pig Ileum Myenteric Plexus ..................... 172
3. Central Nervous System ............................... 173
4. Agonist Regulation of G Protein Levels. . . . . . . . . . . . . . . . .. 173
V. Effector System Function May Be Enhanced
After Sustained Opiate Drug Treatment .................... 174
1. Guinea Pig Ileum Myenteric Plexus ..................... 175
2. Neuroblastoma X Glioma (NG 108-15) Hybrid Cells ....... 175
3. Dorsal Root Ganglion-Spinal Cord Cultures. . . . . . . . . . . . .. 176
4. Locus Ceruleus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 177
5. Summary ............................................ 178
VI. Summary: G Proteins and Opioid Tolerance and Dependence.. 180
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 180

CHAPTER 9
Opioid Receptor-Coupled Second Messenger Systems
S.R. CHILDERS. With 3 Figures ................................... 189
A. Introduction ................................................ 189
B. G Protein Coupling to Receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 190
I. General G Protein Structure and Function. . . . . . . . . . . . . . . . .. 190
II. Opioid Receptors Are Coupled to G Proteins ............... 193
C. Opioid-Inhibited Adenylyl Cyclase ............................. 194
I. Acute Effects of Opioid Agonists on Adenylyl Cyclase
in Transformed Cell Lines . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 195
II. Acute Effects of Opioid Agonists on Adenylyl Cyclase
in Brain. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 196
III. Chronic Effects of Opioid Agonists ........................ 198
IV. Biological Roles for Opioid-Inhibited Adenylyl Cyclase ....... 201
D. Other Second Messenger Systems .............................. 203
xx Contents

I. Stimulation of Adenylyl Cyclase . . . . . . . . . . . . . . . . . . . . . . . . . .. 203

II. Cyclic GMP ............................................ 204
III. Phosphatidylinositol Turnover and Effects
on Membrane Lipids. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 204
IV. Opioid-Dependent Protein Phosphorylation ................. 206
E. Conclusions ................................................ 207
References ................................................... " 208

CHAPTER 10
Allosteric Coupling Among Opioid Receptors:
Evidence for an Opioid Receptor Complex
R.B. ROTHMAN, l.W. HOLADAY, and F. PORRECA. With 4 Figures ...... 217
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 217
B. Evidence for a ll-o-0pioid Receptor Complex ................... 217
I. Ligand-Binding Data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 217
1. Evidence that Il-Ligands Noncompetitively Inhibit
o-Receptor Binding ................................... 217
2. Evidence that o-Ligands Noncompetitively Inhibit
Il-Receptor Binding ................................... 220
II. o-Agonist - Il-Agonist Interactions ......................... 223
1. Early Studies: Analgesia Model. . . . . . . . . . . . . . . . . . . . . . . .. 223
2. More Recent Studies: Analgesia Model ................ " 224
III. Il-Antagonist - o-Antagonist Interactions ................... 228
IV. Linkage Studies . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 229
C. Evidence for a K-Binding Site Associated
with the ll-o-0pioid Receptor Complex. . . . . . . . . . . . . . . . . . . . . . . .. 230
I. In Vitro, Electrophysiological, Anatomical,
and Biochemical Evidence 231
for a ll-o-0pioid Receptor Complex ....................... .
D. Conclusions ................................................ 232
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 233

Section B: Chemistry of Opioids with Alkaloid Structure

CHAPTER 11
Chemistry of Nonpeptide Opioids
S. ARCHER .......... " ......................................... 241
A. Introduction ................................................ 241
B. Biosynthesis of Morphine, Codeine, and Thebaine ............. " 241
C. Morphine and Its Companions ................................ 244
D. Transformation Products of Thebaine ........................ " 247
Contents XXI

E. Morphinans ................................................ 251

F. Diene Adducts Derived from Thebaine. . . . . . . . . . . . . . . . . . . . . . . .. 255
G. 6,7-Benzomorphans .......................................... 257
H. Piperidine-Based Opioids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 262
I . Ethylene Diamines ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 265
J . Acyclic Opioids .... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 269
K. Concluding Remarks ......................................... 270
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 271

CHAPTER 12
Selective Nonpeptide Opioid Antagonists
P.S. PORTOGHESE. With 8 Figures .................................. 279
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 279
B. Receptor Selectivity ......................................... 280
C. Il-Seiective Opioid Antagonists. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 281
D. 8-Selective Opioid Antagonists . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 282
E. K-Selective Opioid Antagonists. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 288
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 291

CHAPTER 13
Presence of Endogenous Opiate Alkaloids in Mammalian Tissues
S. SPECTOR and J. DONNERER. With 3 Figures ....................... 295
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 295
B. Technical Principles Used in the Isolation of Alkaloid Compounds
from Animal Tissue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 295
C. Identification of Endogenous Opiate Alkaloids
in Mammalian Tissue ........................................ 297
D. Biosynthesis of Mammalian Morphine .......................... 298
E. Regulation of Endogenous Morphine and Search
for a Physiological Role ., . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 301
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 302

Section c: Opioid Peptides

CHAPTER 14
Regulation of Opioid Peptide Gene Expression
V. HOLLT. With 2 Figures ........................................ 307
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 307
B. Structure and Regulatory Elements of the Opioid Peptide Genes. .. 308
I. Proopiomelanocortin..................................... 308
XXII Contents

II. Proenkephalin .......................................... 310

III. Prodynorphin ........................................... 312
C. Gene Regulation ............................................ 313
I. Proopiomeianocortin..................................... 313
1. Adenohypophysis.................................... 313
2. Intermediate Pituitary ................................ 316
3. Hypothalamus ....................................... 318
4. Peripheral Tissues ................................... 318
5. Tumors ............................................. 319
II. Proenkephalin .......................................... 320
1. Striatum............................................ 320
2. Hypothalamus....................................... 321
3. Hippocampus and Cortex ............................. 322
4. Spinal Cord and Lower Brainstem. . . . . . . . . . . . . . . . . . . . .. 323
5. Pituitary ............................................ 323
6. Adrenal Medulla .................................... 324
7. Heart .............................................. 326
8. Gonads............................................. 327
9. Immune System ..................................... 327
10. Cell Lines ........................................... 328
Ill. Prodynorphin ........................................... 328
1. Hypothalamus ....................................... 328
2. Striatum............................................ 329
3. Hippocampus ....................................... 330
4. Spinal Cord ......................................... 331
5. Pituitary ............................................ 331
6. Peripheral Tissues ................................... 332
D. Summary................................................... 332
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 333

CHAPTER 15
Regulation of Pituitary Proopiomelanocortin Gene Expression
l.L. ROBERTS, N. LEVIN, D. LORANG, l.R. LUNDBLAD, S. DERMER,
and M. BLUM. With 2 Figures .................................... 347
A. Introduction................................................ 347
I. The POMC Gene ........................................ 347
II. Intracellular Processes Regulating POMC Secretion .......... 349
B. Proopiomeianocortin mRNA Levels in Pituitary . . . . . . . . . . . . . . . .. 351
I. Whole Animal Studies ................................... 352
1. Adrenalectomy....................................... 352
2. Hypothalamic Factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 353
3. Intermediate Lobe POMC mRNA Levels ................ 354
II. In Vitro Systems ........................................ 355
Contents XXIII

1. Glucocorticoids....................................... 356
2. cAMP- and Calcium-Dependent Processes ............... 356
III. Summary............................................... 358
C. Proopiomelanocortin Gene Transcription . . . . . . . . . . . . . . . . . . . . . .. 358
I. Modulation of POMC hnRNA Levels . . . . . . . . . . . . . . . . . . . . .. 359
II. Whole Animal Studies ................................... 360
III. Primary and AtT20 Cell Culture. . . . . . . . . . . . . . . . . . . . . . . . . .. 361
IV. Summary............................................... 363
D. Regulatory Elements in the POMC Gene ....................... 364
I. Basal and Tissue-Specific Promoter Elements. . . . . . . . . . . . . . .. 364
II. Glucocorticoid Regulatory Elements . . . . . . . . . . . . . . . . . . . . . .. 367
III. Promoter Elements and Second Messenger Pathways. . . . . . . .. 368
IV. Summary ............................................... 369
E. Conclusions ................................................ 370
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 341

CHAPTER 16
Molecular Mechanisms in Proenkephalin Gene Regulation
N. KLEY and J.P. LOEFFLER ....................................... 379
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 379
B. Cellular Signaling Pathways Mediating PENK Gene Induction ..... 380
I. Membrane Associated Events and Second Messengers. . . . . . .. 380
1. Regulation of PENK Gene Expression by Electrical Activity
and Ca 2 + Metabolism in Excitable Cells. . . . . . . . . . . . . . . . .. 380
2. Cyclic AMP as a Regulator of PENK Gene Expression. . . .. 381
3. Phosphoinositide Hydrolysis and PENK Gene Regulation .. 381
II. Regulation of PENK Gene Expression by Third Messengers ... 382
C. Mechanisms of PENK Gene Transcriptional Regulation .......... 383
I. Transcriptional Regulation of the Endogenous PENK Gene . .. 384
II. Gene Transfer Approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 385
III. DNA-Responsive Elements ............................... 387
D. Summary ................................................... 389
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 390

CHAPTER 17
Proopiomelanocortin Biosynthesis, Processing and Secretion:
Functional Implications
E. YOUNG, D. BRONSTEIN, and H. AKIL. With 1 Figure ............... 393
A. Introduction................................................ 393
B. Tissue-Specific Processing .................................... 394
I. Anterior Lobe .......................................... 394
II. Intermediate Lobe ....................................... 396
XXIV Contents

III. Brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 397

C. Proopiomelanocortin Processing and Modifying Enzymes . . . . . . . .. 400
D. Possible Functional Significance of Posttranslational Modifications
to POMC-Derived Peptides ................................... 401
I. Anterior Lobe .......................................... 403
II. Intermediate Lobe. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 404
III. Brain . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 405
1. Central Analgesia, Tolerance and Dependence. . . . . . . . . . .. 405
2. Reinforcement ....................................... 408
3. Autonomic Functions ................................. 409
IV. Immune System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 410
E. Conclusion ................................................. 412
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 413

CHAPTER 18
Biosynthesis of Enkephalins and Proenkephalin-Derived Peptides
J. ROSSlER. With 5 Figures ....................................... 423
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 423
B. History .................................................... 424
C. Enkephalin Biosynthesis in the Adrenal Medulla. . . . . . . . . . . . . . . .. 425
D. Molecular Biology ........................................... 431
E. Enkephalin Biosynthesis in the CNS ........................... 433
F. Synenkephalin .............................................. 436
G. Molecular Evolution of Proenkephalin ......................... 437
H. Extraneuronal Proenkephalin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 438
I. Reproductive Tissue . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 438
II. Glial Cells. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 438
III. Immune System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 439
I. Processing of Proenkephalin .................................. 439
J. Regulation................................................. 440
K. Conclusion ................................................. 441
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 441

CHAPTER 19
Prodynorphin Biosynthesis and Posttranslational Processing
R. DAY, K.A. TRUJILLO, and H. AKIL. With 2 Figures ............... 449
A. History of Dynorphin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 449
B. Posttranslational Processing Signals ............................ 450
C. Prodynorphin Biosynthesis and Processing in Peripheral Tissues ... 451
D. Processing Pathway of Prodynorphin ........................... 453
Contents XXV

E. Functional Significance of Prodynorphin Peptide Processing . . . . . .. 458

I. Striatonigral System ..................................... 461
II. Other Systems .......................................... 463
F. Conclusions ................................................ 463
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 463

CHAPTER 20
Anatomy and Function of the Endogenous Opioid Systems
H. KHACHATURIAN, M.K.H. SCHAFER, and M.E. LEWIS.
With 1 Figure ................................................ " 471
A. Introduction ................................................ 471
B. Immunocytochemical Anatomy of Opioid Systems ............. " 472
I. Proopiomelanocortin..................................... 472
II. Pro en kephalin .......................................... 473
III. Prodynorphin ......................................... " 474
C. In Situ Hybridization Histochemical Studies ................... " 475
I. Proopiomelanocortin mRNA . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 476
II. Proenkephalin and Prodynorphin mRNA ................. " 477
III. Expression of Opioids in Nonneuronal Cells ................. 481
D. Opioid Receptors and Functional Systems ...................... 482
I. Problems in the Functional Analysis
of Endogenous Opioid Systems. . . . . . . . . . . . . . . . . . . . . . . . . . .. 482
II. Opioid Peptide-Receptor Relationships ..................... 483
E. Functional Roles of Opioid Systems .......................... " 484
I. Endogenous Pain Control Systems. . . . . . . . . . . . . . . . . . . . . . . .. 484
II. Extrapyramidal Motor Systems . . . . . . . . . . . . . . . . . . . . . . . . . . .. 486
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 488

CHAPTER 21
Atypical Opioid Peptides
H. TESCHEMACHER .............................................. 499
A. Introduction .............................................. " 499
I. Atypical Representatives of Natural Opioid Peptides
(Atypical Natural Opioid Peptides) ........................ 499
II. Peptides with Indirect Opioid or Opioid Antagonist Activity. " 499
B. Atypical Opioid Peptides ..................................... 501
I. Structure and Activity .................................... 501
1. a-Casein Exorphins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 512
2. ~-Casomorphins . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 512
3. ~-Casorphin, a- and ~-Lactorphins ...................... 513
XXVI Contents

4. Hemorphins and Cytochrophins. . . . . . . . . . . . . . . . . . . . . . . .. 513

5. Dermorphins and Deltorphins .......................... 513
II. Origin and Destination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 515
1. Milk Protein-Derived Opioid Peptides ................... 515
2. Hemoglobin- or Cytochrome b-Derived Opioid Peptides ... 516
3. Amphibian Skin Protein-Derived Opioid Peptides ......... 517
C. Opioid Antagonists Sharing Characteristics
with Atypical Opioid Peptides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 517
I. Structure and Activity ................................... , 518
1. Casoxins............................................. 518
2. Lactoferroxins........................................ 518
II. Origin and Destination .................................. , 518
D. Atypical Opioid Peptide Analogues with Agonist
or Antagonist Activity ....................................... 518
I. Agonists ............................................... 521
1. Il-Selective Opioid Receptor Ligands .................... 521
2. o-Selective Opioid Receptor Ligands .................... 521
II. Antagonists............................................. 521
E. Concluding Remarks. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 521
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 522

CHAPTER 22
Opioid Peptide Processing Enzymes
L.D. FRICKER . . . . . . . . . , .... , . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 529
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 529
B. Enzymes in the Endoplasmic Reticulum and Golgi Apparatus ..... 531
I. Signal Peptidase ......................................... 531
II. Glycosylation, Sulfation, and Phosphorylation. . . . . . . . . . . . . .. 531
C. Enzymes in the Secretory Granules ............................ 533
I. Endopeptidases Selective for Paired Basic Residues . . . . . . . . .. 534
II. Opioid Peptide Processing Endopeptidases Selective
for Single Basic Residues . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 536
III. Carboxypeptidase E . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 537
IV. Aminopeptidase B-Like Enzyme .......................... 539
V. Amidation.............................................. 539
VI. Acetylation............................................. 540
D. Extracellular Opioid Peptide Processing Enzymes ................ 541
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 541
Contents XXVII

CHAPTER 23
Peptidase Inactivation of Enkephalins:
Design of Inhibitors and Biochemical, Pharmacological
and Clinical Applications
B.P. ROQUES, A. BEAUMONT, V. DAUGE, and M.-C. FOURNIE-ZALUSKI.
With 2 Figures ................................................. 547
A. Introduction................................................ 547
B. Enkephalin Degrading Enzymes ............................... 548
I. Metabolism of Opioid Peptides . . . . . . . . . . . . . . . . . . . . . . . . . .. 548
II. Substrate Specificity of NEP and APN . . . . . . . . . . . . . . . . . . . .. 550
III. Assays of NEP and APN Activities. . . . . . . . . . . . . . . . . . . . . . .. 551
C. Structure and Molecular Biology of NEP ....................... 551
I. Structure of NEP ....................................... 551
II. Human NEP (CALLA) Gene ............................ 553
D. Localization of Neutral Endopeptidase 24.11 .................... 553
I. Central Nervous System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 553
II. Localization of NEP in Peripheral Tissues. . . . . . . . . . . . . . . . .. 555
III. In Vitro and In Vivo Studies of Enkephalin Degradation
by NEP and APN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 555
E. Inhibitor Design and Synthesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 556
I. Design of Selective and Mixed Inhibitors
of Neutral Endopeptidase 24.11 and Aminopeptidase N . . . . .. 556
II. Thiol Inhibitors. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 557
III. Carboxyl Inhibitors ..................................... 559
IV. Hydroxamic Acids and Derivatives ........................ 561
V. Phosphorus-Containing Inhibitors ......................... 563
VI. Aminopeptidase-N and Dipeptidyl Peptidase Inhibitors . . . . .. 563
VII. Development of Mixed Inhibitors
of Enkephalin-Degrading Enzymes ........................ 563
F. Pharmacological Studies of Enkephalin-Degrading-Enzyme
Inhibitors .................................................. 564
I. Inhibitor-Induced Analgesia .............................. 565
II. Inhibitor-Induced Spinal Antinociception .................. 566
III. Peptidase Inhibitors in Chronic Pain. . . . . . . . . . . . . . . . . . . . . .. 567
IV. Tolerance, Dependence, and Side Effects of Selective
and Mixed Inhibitors of NEP and APN . . . . . . . . . . . . . . . . . . .. 567
V. Gastrointestinal Effects .................................. 568
VI. Role of Neutral Endopeptidase-24.11 in Airways. . . . . . . . . . .. 569
VII. Behavioral Effects of Inhibitors ........................... 569
XXVIII Contents

G. Inhibition of NEP Inactivation of Atrial Natriuretic Peptide:

Pharmacological and Clinical Implications ...................... 570
H. Clinical Applications of Selective and Mixed Zn Metallopeptidase
Inhibitors .................................................. 571
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 572

CHAPTER 24
Coexistence of Opioid Peptides with Other Neurotransmitters
R. ELDE and T. HOKFELT. With 6 Figures .......................... 585
A. Principles .................................................. 585
I. Introduction........................................... 585
II. Subcellular Features. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 588
1. Classical Neurotransmitters and Small Synaptic Vesicles .. 589
2. Neuropeptides and Large Granular Vesicles ............. 590
III. Methods for Establishing Coexistence ..................... 590
B. Coexistence Within Areas of the Nervous System . . . . . . . . . . . . . . .. 592
I. Retina................................................ 593
II. Telencephalon......................................... 601
III. Diencephalon ......................................... 602
IV. Mesencephalon........................................ 603
V. Pons and Medulla. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 603
VI. Cerebellum ........................................... 606
VII. Spinal Cord ........................................... 606
VIII. Peripheral Nervous System. . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 607
1. Primary Afferent Neurons. . . . . . . . . . . . . . . . . . . . . . . . . . .. 607
2. Autonomic Ganglion Cells and Their Fibers. . . . . . . . . . . .. 608
3. Adrenal Medulla .................................... 609
4. Enteric Nervous System .............................. 609
C. Implications ................................................ 6U
I. Patterns of Expression. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 611
II. Pharmacology and Physiology ........................... 612
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 612

CHAPTER 25
Interrelationships of Opioid, Dopaminergic, Cholinergic
and GABAergic Pathways in the Central Nervous System
P.L. WOOD. With 2 Figures ...................................... 625
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 625
B. Cholinergic Systems ......................................... 625
I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 625
II. Septohippocampal Cholinergic Pathway .................... 625
Contents XXIX

III. Nucleus Basalis-Cortical Cholinergic Pathway .. . . . . . . . . . . . .. 626

C. Dopaminergic Pathways. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 628
I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 628
II. Nigrostriatal Pathway .................................... 629
III. Mesolimbic Pathways .................................... 632
IV. Mesocortical Pathways ................................... 633
D. GABAergic Pathways ........................................ 634
E. Striatal Opioid Peptide Gene Expression ....................... 635
I. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 635
II. Met- Enkephalin ........................................ 635
III. Dynorphin ............................................. 637
F. Conclusions ................................................ 637
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 638

CHAPTER 26
Selectivity of Ligands for Opioid Receptors
A.D. CORBETI, S.l. PATERSON, and H.W. KOSTERLITZ ................ 645
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 645
B. Methods Used to Determine the Selectivity of Opioid Compounds .. 645
I. Radioreceptor Binding Assays ............................ 646
II. Bioassays ............................................... 647
C. Selectivity of Endogenous Opioid Peptides . . . . . . . . . . . . . . . . . . . . .. 648
I. Proenkephalin-Derived Peptides ........................... 650
1. Activity in Binding Assays ... . . . . . . . . . . . . . . . . . . . . . . . . .. 650
2. Activity in Bioassays ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 650
II. Prodynorphin-Derived Peptides ........................... 651
1. Activity in Binding Assays ............................. 651
2. Activity in Bioassays .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 653
III. Proopiomelanocortin-Derived Peptides ..................... 653
1. Activity in Binding Assays ... . . . . . . . . . . . . . . . . . . . . . . . . .. 653
2. Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 655
IV. Dermorphin and Deltorphins ............................. 655
1. Activity in Binding Assays .. . . . . . . . . . . . . . . . . . . . . . . . . . .. 655
2. Activity in Bioassays ... . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 657
D. Selectivity of Nonendogenous Opioid Compounds ............... 657
I. Compounds with a Preference for the Il-Binding Site ......... 660
1. Activity in Binding Assays . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 660
2. Agonist Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . . . . .. 662
3. Antagonist Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . .. 662
II. Compounds with a Preference for the o-Binding Site ......... 663
1. Activity in Binding Assays .... . . . . . . . . . . . . . . . . . . . . . . . .. 663
2. Agonist Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . . . . .. 668
3. Antagonist Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . .. 668
xxx Contents

Ill. Compounds with a Preference for the K-Binding Site ......... 669
1. Activity in Binding Assays .. . . . . . . . . . . . . . . . . . . . . . . . . . .. 669
2. Agonist Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . . . . .. 669
3. Antagonist Activity in Bioassays . . . . . . . . . . . . . . . . . . . . . . .. 669
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 673

CHAPTER 27
Development of Receptor-Selective Opioid Peptide
Analogs as Pharmacologic Tools and as Potential Drugs
P.W. SCHILLER. With 2 Figures ................................... 681
A. Introduction................................................ 681
B. Determination of Receptor Selectivity. . . . . . . . . . . . . . . . . . . . . . . . .. 683
C. Development of 11-, 0-, and K-Receptor-Selective Opioid Peptide
Analogs with Agonist Properties. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 684
I. I1-Selective Agonists ..................................... 684
1. Linear Opioid Peptide Analogs . . . . . . . . . . . . . . . . . . . . . . . .. 684
2. Opioid Peptide Dimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 688
3. Cyclic Opioid Peptide Analogs .......................... 689
II. o-Selective Agonists ..................................... 692
1. Linear Opioid Peptide Analogs . . . . . . . . . . . . . . . . . . . . . . . .. 692
2. Opioid Peptide Dimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 696
3. Cyclic Opioid Peptide Analogs. . . . . . . . . . . . . . . . . . . . . . . . .. 696
III. K-Selective Agonists ..................................... 697
D. Selective Opioid Peptide Analogs with Antagonist Properties. . . . .. 697
E. Irreversible Opioid Receptor Peptide Ligands . . . . . . . . . . . . . . . . . .. 699
1. Chemical Affinity Labels .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 699
II. Photoaffinity Labels ..................................... 699
F. Selective Opioid Peptide Analogs as Drug Candidates ............ 701
G. Conclusions ................................................ 703
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 704

CHAPTER 28
Ontogeny of Mammalian Opioid Systems
J.E. PINTAR and R.E.M. SCOTT ................................... 711
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 711
B. Embryological Considerations ................................. 711
C. Opioid Gene Activation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 712
I. Proopiomelanocortin..................................... 712
1. Brain ............................................... 712
2. Pituitary............................................. 713
3. Testis ............................................... 714
4. Placenta ............................................. 714
Contents XXXI

II. Enkephalin ............................................. 714

1. Brain (Striatal) .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 715
2. Glia ................................................. 715
3. Fetal Mesoderm ...................................... 716
III. Dynorphin ............................................. 716
D. Ontogeny of Opioid Precursor Processing. . . . . . . . . . . . . . . . . . . . . .. 717
I. Proopiomelanocortin..................................... 717
1. Immunocytochemical Analyses ......................... 717
2. Biochemical Analyses ................................. 718
II. Dynorphin ............................................. 718
E. Ontogeny of Regulated Release . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 719
I. Secretory Granules, Regulators of POMC Secretion
and the Portal System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 719
II. Functional Receptors for Secretagogues .................... 720
F. Function ................................................... 721
I. Ontogeny of Opioid Receptors . . . . . . . . . . . . . . . . . . . . . . . . . . .. 721
II. Putative Role(s) of Opioid Peptides
in Developmental Processes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 723
G. Prospectus.................................................. 723
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 724

Section D: Neurophysiology

CHAPTER 29
Opioids and Sensory Processing in the Central Nervous System
A.W. DUGGAN and S.M. FLEETWOOD-WALKER. With 4 Figures 731
A. Introduction ................................................ 731
B. Opioids and the Spinal Cord .................................. 731
I. Spinal Processing of Nociceptive Information ................ 731
II. Systemic Administration of Opiates and the Responses
of Spinal Neurones ...................................... 732
1. Neuronal Types ...................................... 733
2. Responses to Peripheral Stimuli. . . . . . . . . . . . . . . . . . . . . . . .. 733
III. Localized Administration of Opioids . . . . . . . . . . . . . . . . . . . . . .. 736
1. I!-Receptor-Preferring Ligands ........................ " 737
2. o-Receptor-Preferring Ligands ........................ " 740
3. K-Receptor-Preferring Ligands .......................... 741
IV. Functional Consequences of Opioid Receptor Activation
to Spinal Sensory Processing . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 743
1. Opioid Receptors and the Central Terminals of Nociceptors 743
2. Receptors on the Somata and Processes of Spinal Neurones 745
3. Receptors and Supraspinal Fibres ....................... 747
XXXII Contents

V. Opiates and Descending Inhibition ......................... 748

VI. Physiological Roles of Opioid Peptides in Sensory Processing .. 749
1. Spinal Release of Opioid Peptides . . . . . . . . . . . . . . . . . . . . . .. 750
2. Tonic Opioidergic Inhibition ........................... 750
3. Phasic Opioidergic Inhibition ........................... 752
C. Thalamus and Cerebral Cortex . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 753
I. Thalamus .............................................. 753
1. Ventrobasal Nuclei .................................... 753
2. Medial and Dorsal Thalamic Nuclei ..................... 755
II. Cerebral Cortex ......................................... 757
D. Deficits in Knowledge and Prospects for Future Research ......... 758
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 760

CHAPTER 30
Opioid Actions on Membrane Ion Channels
R.A. NORTH. With 4 Figures ..................................... 773
A. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 773
B. Calcium Channels ........................................... 774
I. Types of Calcium Channels .............................. 774
II. Il-Receptors ........................................... 775
III. (i-Receptors ........................................... 775
IV. K-Receptors ........................................... 777
V. Unclassified Receptors .................................. 777
VI. Experiments on Action Potential Duration ................ 777
VII. Type of Calcium Current Inhibited ....................... 778
VIII. Mechanism of Opioid Action . . . . . . . . . . . . . . . . . . . . . . . . . . .. 778
1. Role of G Proteins .................................. 778
2. Time Course of Agonist Action ....................... 779
3. Single Channel Studies ............................... 780
4. Voltage Dependence of Agonist Action? ............... 780
IX. Other Receptors That Reduce Calcium Currents .. . . . . . . . .. 780
X. Calcium Current Inhibition and Presynaptic Inhibition ...... 781
C. Potassium Channels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 781
I. Types of Potassium Channels. . . . . . . . . . . . . . . . . . . . . . . . . . .. 781
II. Il-Receptors ........................................... 782
III. (i-Receptors ........................................... 782
IV. Other Receptors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 784
V. Experiments on Action Potential Duration ................ 784
VI. Hyperpolarization and Inhibition of Firing ................. 784
VII. Type of Potassium Current Increased ..................... 784
VIII. Mechanism of Opioid Action ............................ 787
Contents XXXIII

1. Role of G Proteins .................................. 787

2. Time Course of Action. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 788
3. Single Channel Studies ............................... 788
IX. Other Receptors That Increase Potassium Conductance . . . .. 790
X. Potassium Conductance Increase and Presynaptic Inhibition 790
D. Other Ion Channels .......................................... 791
E. Changes in Tolerance and Dependence. . . . . . . . . . . . . . . . . . . . . . . .. 791
F. Concluding Remarks ......................................... 792
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .. 793

Subject Index .................................................. 799

Contents of Companion Volume 104, Part II

Section E: Pharmacology and Behavior

CHAPTER 31
Brainstem Mechanisms of Pain Modulation: Anatomy and Physiology
H.L. FIELDS
CHAPTER 32
Supraspinal Opioid Receptors in Antinociception. F. PORRECA and T .F. BURKS
CHAPTER 33
The Spinal Actions of Opioids. T. L. Y AKSH
CHAPTER 34
Peripheral Mechanisms of Opioid Analgesia. C. STEIN
CHAPTER 35
Acupuncture and Stimulation-Produced Analgesia. lI-SHENG HAN
CHAPTER 36
Multiple Opioid Systems and Chronic Pain. M.l. MILLAN
CHAPTER 37
Gastrointestinal Effects of Opioids. W. KROMER
CHAPTER 38
Role of Endogenous Opioids in Central Cardiovascular Regulation
and Dysregulation. A.1. FADEN
CHAPTER 39
Physiological Functions of Opioids: Temperature Regulation.
M.W. ADLER and E.B. GELLER
CHAPTER 40
Opioid Mechanisms in the Control of Food Consumption
and Taste Preferences. S.l. COOPER and T.e. KIRKHAM
CHAPTER 41
Opioids in Respiration and Vomiting. 1. FLOREZ and M.A. HURLE
CHAPTER 42
Opioid Systems and Stress. R. PRZEWLOCKI
XXXVI Contents of Companion Volume 104, Part II

CHAPTER 43
Role of Endogenous Opioids and Opioid Receptors in the Central Nervous
System Injury. A.I. FADEN
CHAPTER 44
Opioids: Epilepsy and Neuroprotection. F.C. TORTELLA
CHAPTER 45
Opioids in Immunologic Processes. H.U. BRYANT and J.W. HOLADAY
CHAPTER 46
Effects of Opioids on the Spontaneous Behavior of Animals. A. Cow AN
CHAPTER 47
Involvement of Opioid Peptides in Learning and Memory. J. L. MCGAUGH,
I.B. INTROINI-COLLISON, and C. CASTELLANO
CHAPTER 48
Correlations Between the Pharmacodynamic Characteristics
of Opioid Agonists and Their Behavioral Effects.
A.J. BERTALMIO, c.P. FRANCE, and J.H. WOODS

Section F: Neuroendocrinology

CHAPTER 49
Opioid Peptides in the Regulation of Anterior Pituitary Hormones.
S.G. CELLA, V. LOCATELLI, and E.E. MULLER
CHAPTER 50
Opioids and the Neuroendocrine Control of Reproduction. O.F.X. ALMEIDA
CHAPTER 51
Opioids in the Neurohypophysial System. R.J. BICKNELL
CHAPTER 52
Opioid Peptide Expression in Peripheral Tissues
and Its Functional Implications. D.L. KILPATRICK

Section G: Reinforcement - Tolerance/Dependence

CHAPTER 53
Opioid Tolerance and Physical Dependence and Their Relationship.
E.L. WAY
Contents of Companion Volume 104, Part II XXXVII

CHAPTER 54
Opioid Tolerance/Dependence in Isolated Organs. R. SCHULZ
CHAPTER 55
Opioid Abuse Liability Assessment in Rhesus Monkeys. J.H. WOODS,
c.P. FRANCE, G. WINGER, A.J. BERTALMIO, K. SCHWARZ-STEVENS
CHAPTER 56
Motivational Effects of Opioids. T.S. SHIPPEN BERG

Section H: Clinical Aspects

CHAPTER 57
CSF Opioids in Pathophysiology. F. NYBERG
CHAPTER 58
Circulating Opioids in Man. L. McLoUGHLIN, S. MEDBACK,
and A.B. GROSSMAN
CHAPTER 59
Opioid Analgesics in Clinical Pain Management. K.M. FOLEY
CHAPTER 60
Opioids in Operative Anesthesia. P.L. BAILEY
CHAPTER 61
p-Endorphin in Human Reproduction. F. PETRAGLIA, G. COMITINI,
and A.R. GENAZZANI
CHAPTER 62
Opioids in the Etiology and Treatment of Psychiatric Discorders.
D. NABER

CHAPTER 63
Opioid Addiction. c.P. O'BRIEN

Subject Index