Table Of ContentCorticotropin-Releasing
Factor:
Basic and Clinical Studies
of a
Neuropeptide
Editors
Err01 B. De Souza
Chief, Neurobiology Laboratory
National Institute on Drug Abuse
Addiction Research Center
Baltimore, Maryland
Charles B. Nemeroff
Professor
Department of Psychiatry
Division of Biological Psychiatry
Duke University Medical Center
Durham. North Carolina
Boca Raton London New York
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Library of Congress Cataloging-in-Publication Data
Corticotropin-releasing factor.
Includes bibliographies and index.
1. Corticotropin releasing hormone. I. De Souza,
Errol B. II. Nemeroff, Charles B. [DNLM: 1. Cortico-
tropin Releasing Hormone. WK 515 C8293]
QP572.C62C67 1990 599’ .019’2456 89-9933
ISBN 0-8493-4550-2
A Library of Congress record exists under LC control number: 89009933
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FOREWORD
The chemical and biological characterization of corticotropin-releasing factor (CRF) has
evolved from two fundamental concepts developed over 30 years ago. The investigations
of Scharrer and Scharrer, Harris, and others supported the theory that factors secreted by
hypothalamic neurons into the hypothalamo-hypophysial portal vessel system could then
reach and modulate the functions of the anterior pituitary gland. Evidence for the existence
of a hypothalamic factor controlling ACTH secretion was provided by the in vivo studies
of Harris, Hume, Fortier, and others and by the in vitro experiments of Guillemin and Saffron
and Schally. The other key concept, derived from the work of Bernard, Cannon, and Selye,
is that when exposed to real or perceived challenge to its internal homeostasis, an organism
reacts with an array of hormonal, autonomic, and behavioral responses. Among the principle
"stress responses" is the activation of the pituitary-adrenal cortical axis.
Although biologically recognized in the 1950s, the chemical identity of CRF was not
elucidated until 1981 when our group reported the primary sequence of ovine CRF, a 41
amino acid straight-chain polypeptide which is a highly potent stimulator of the biosynthesis
and secretion of ACTH and other proopiomelanocortin (POMC) derived peptides such as
P-endorphin. The development of improved cell culture-based bioassays, the recognition
and identification of some of the other substances including vasopressin and epinephrine in
hypothalamic extracts capable of releasing ACTH, and the improvement of purification and
sequencing techniques contributed to the success of this effort. The following year, the
structures of rat CRF isolated by us from rat hypothalamus and human CRF deduced by
Numa's group using human genomic clones were found to be identical to one another. More
recently, the sequences of bovine, carpine, and porcine CRF were determined.
High affinity CRF binding sites, and cellular actions of CRF have been described for
tissues in which the peptide has been localized. CRF activates adenylate cyclase and modifies
firing rates in several brain regions in which receptors have been identified by membrane
binding and autoradiographic techniques.
In addition to activation of the pituitary-adrenal axis, stressful stimuli evoke many other
endocrine, autonomic, and behavioral responses. The intracerebroventricular administration
of CRF provokes several stress-like responses including activation of the sympathetic nervous
system and inhibition of the parasympathetic nervous system with consequential increases
in heart rate, blood pressure, blood sugar, and inhibition of gastrointestinal functions. Fur-
thermore, elevating CRF levels in the brain inhibits the secretion of gonadotropins and
growth hormone by modulating the production of gonadotropin-releasing hormone and so-
matostatin. Finally, significant effects of CRF on behavior have been noted and many are
consistent with increased arousal and emotional reactivity to the environment. Recent ex-
periments using CRF antagonists support the hypothesis that endogenous CRF participates
in many of the neuroendocrine, autonomic, and behavioral responses to stress. These ob-
servations along with evidence for increased production of CRF in certain human affective
disorders as revealed by inappropriate pituitary-adrenal activation or elevated CRF levels in
the cerebrospinal fluid have raised questions concerning the role of the peptide in conditions
such as anorexia nervosa and depression. Future fundamental work on the structure of the
CRF receptor(s), on the development of improved CRF receptor antagonists for physiologic
and clinical studies, on the functions of brain CRF pathways in behavior and in coordinating
the stress response may yield improved pharmacologic tools and provide insight concerning
the nature of stress-related diseases.
Wylie Vale
Clayton Foundation Laboratories for Peptide Biology
The Salk Institute for Biological Studies
La Jolla, California
THE EDITORS
Err01 B. De Souza, Ph.D., is Chief, Neurobiology Laboratory, Addiction Research
Center, National Institute on Drug Abuse, Alcohol, Drug Abuse and Mental Health Admin-
istration, Baltimore, Maryland. He also holds a joint appointment as Assistant Professor of
Pathology, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Dr. De Souza graduated in 1976 from the University of Toronto with a B.A. degree in
Physiology and obtained his Ph.D. degree in 1982 from the Department of Physiology,
University of Toronto, Toronto, Canada. Subsequently, in 1983, he spent 2 years in the
Department of Neuroscience at Johns Hopkins University School of Medicine where he
carried out extensive work imaging receptors using autoradiographic techniques.
Dr. De Souza is a member of the American Association for the Advancement of Science,
the Endocrine Society, the International Brain Research Organization, and the Society for
Neuroscience.
Dr. De Souza has presented over 40 invited lectures and seminars at national and
international meetings and at universities and institutes. He has published approximately 90
research papers and 100 abstracts. His current major research interests have focused on the
identification, characterization, and localization of various neurotransmitter receptors in brain
and in the periphery. In addition, he has carried out extensive studies examining changes
in receptors and receptor-mediated second messengers in neurodegenerative diseases and
neuropsychiatric disorders.
Charles B. Nerneroff, M.D., Ph.D., was born in New York City almost 40 years ago
and was educated in the New York City Public School System. After graduating from the
City College of New York in 1970, he briefly enrolled as a graduate student in evolutionary
biology at the American Museum of Natural History. An interest in neuroscience, however,
led him to a research assistant's position at the Neurochemistry Laboratory at McLean
Hospital in Belmont, Massachusetts. He simultaneously enrolled in graduate school at North-
eastern University and received a Master's degree in Biology in 1973. He then relocated to
North Carolina and enrolled in the Ph.D. program in Neurobiology at the University of
North Carolina at Chapel Hill. His Ph.D. work on thyrotropin-releasing hormone was per-
formed under the direction of a psychiatrist, Arthur J. Prange, Jr. After a year of post-
doctoral training in neurochemistry, he enrolled in medical school at the University of North
Carolina in Chapel Hill. After residency training in psychiatry both at the University of
North Carolina and at Duke University, he joined the faculty of Duke University and is
currently Professor of Psychiatry, Pharmacology and Neurobiology and Chief of the Division
of Biological Psychiatry at Duke University. He has been certified in psychiatry from the
American Board of Psychiatry and Neurology. He has published approximately 260 research
reports and reviews, has nine edited books published or in press, and has been an invited
speaker on more than 200 occasions. He has received a number of awards for his research
including a MERIT Award from the National Institute of Mental Health, the Anna Monika
Prize for Research in Endogenous Depression (1987), the Daniel H. Efron Award of the
American College of Neuropsychopharmacology (1987), the Curt P. Richter Award from
the International Society of Psychoneuroendocrinology (1985), the Jordi Folch-Pi Award
from the American Society for Neurochemistry (1987) and the Judith Silver Memorial Young
Scientist Award from the National Alliance for the Mentally I11 (1988). He is on the editorial
board of ten journals. Finally, he would, on most days, prefer to be at the seashore with
his wife, Melissa, and two children, Matthew and Amanda.
CONTRIBUTORS
Greti Aguilera Err01 B. De Souza
Chief, Section on Endocrine Physiology Chief
National Institute of Child Health and Neurobiology Laboratory
Human Development Neuroscience Branch
National Institutes of Health National Institute on Drug Abuse
Bethesda, Maryland Addiction Research Center
Baltimore, Maryland
George Battaglia
James 0. Douglass
Assistant Professor
Assistant Staff Scientist
Department of Pharmacology
Vollum Institute for Advanced
Loyola University of Chicago
Stritch School of Medicine Biomedical Research
Assistant Professor
Maywood, Illinois
Departments of Anatomy and Neurology
Oregon Health Sciences University
Garth Bissette
Portland, Oregon
Assistant Professor and CO-Director
Laboratory of Psychoneuroendocrinology
Cindy L. Ehlers
Duke University Medical Center
Assistant Member
Durham, North Carolina
Department of Neurophmacology
Research Institute of Scripps Clinic
Karen T. Britton
La Jolla, California
Department of Psychiatry
San Diego Veterans Administration
Laurel A. Fisher
Medical Center
Assistant Professor
University of California School of
Department of Pharmacology
Medicine
University of Arizona Health Sciences
La Jolla, California
Center
Tucson, Arizona
Marvin R. Brown
Professor Mirza Flores
Departments of Medicine and Surgery Visiting Fellow
University of California Medical Center National Institute of Child and Human
San Diego, California Development
National Institutes of Health
Pilar Carvallo Bethesda, Maryland
Visiting Fellow
Department of Endocrinology Thackery S. Gray
National Institute of Child Health and Associate Professor
Human Development Department of Anatomy
National Institutes of Health Loyola Stritch School of Medicine
Bethesda, Maryland Maywood, Illinois
Kevin J. Catt Dimitri E. Grigoriadis
Chief Visiting Staff Fellow
Endocrinology and Reproduction Neuroscience Branch
Research Branch National Institute on Drug Abuse
National Institutes of Health Addiction Research Center
Bethesda, Maryland Baltimore, Maryland
Mark M. Gunion Samuel M. McCann
Assistant Research Psychologist Professor and Director, Neuropeptide
Department of Medicine and Brain Division
Research Institute Department of Physiology
Los Angeles, California University of Texas Health Science
Sepulveda VA Medical Center Center
Sepulveda, California Dallas, Texas
James P. Harwood
Monica Millan
Executive Secretary
Staff Fellow
National Institute of Aging
National Institute of Child and Human
National Institutes of Health
Development
Bethesda, Maryland
National Institutes of Health
Bethesda, Maryland
Edward Herbert (Deceased)
Vollum Institute for Advanced
Biomedical Research John E. Morely
Oregon Health Sciences University Professor
Portland, Oregon Department of Medicine
St. Louis University Medical School
Florian Holsboer St. Louis, Missouri
Professor and Chairman
Department of Psychiatry
Charles B. Nemeroff
University of Freiburg
Professor
Freiburg, West Germany
Department of Psychiatry
Division of Biological Psychiatry
Thomas R. Insel
Duke University Medical Center
Senior Staff Scientist
Durham, North Carolina
Laboratory of Clinical Science
National Institute of Mental Health
National Institutes of Health Michael J. Owens
Poolesville, Maryland Postdoctoral Fellow
Department of Psychiatry
Ned H. Kalin Duke University Medical Center
Director of Research Durham, North Carolina
Department of Psychiatry
University of Wisconsin Medical School
Richard E. Powers
Madison, Wisconsin
Assistant Professor
Departments of Psychiatry and Pathology
George F. Koob
University of Alabama School of
Associate Member
Medicine
Department of Neuropharmacology
Birmingham, Alabama
Research Institute of Scripps Clinic
La Jolla, California
Donald L. Price
Allen S. Levine Departments of Pathology, Psychiatry,
Deputy ACOS for Research Neurology, and Neuroscience
Department of Research Johns Hopkins University School of
Veterans Administration Medical Center Medicine
Minneapolis, Minnesota Baltimore, Maryland
Catherine Rivier Yvette Tache
Associate Research Professor Professor in Residence
Clayton Foundation Laboratories for Department of Medicine
Peptide Biology VA Medical Center, West Los Angeles
The Salk Institute University of California at Los Angeles
La Jolla, California Los Angeles, California
Paul E. Sawchenko Robert C. Thompson
Associate Professor Postdoctoral Fellow
Laboratory of Neuronal Structure and Mental Health Research Institute
Function University of Michigan
The Salk Institute Ann Arbor, Michigan
San Diego, California
Wylie Vale
Audrey F. Seasholtz Professor and Head
Assistant Research Scientist Clayton Foundation Laboratories for
Mental Health Research Institute Peptide Biology
University of Michigan The Salk Institute
Ann Arbor, Michigan La Jolla, California
George Robert Siggins
Rita J. Valentino
Department of Neuropharmacology
Assistant Professor
Research Institute of Scripps Clinic
Department of Mental Health Sciences
La Jolla, California
Hahnemann University
Philadelphia, Pennsylvania
Mark Smith
Medical Staff Fellow
Ulrich von Bardeleben
Clinical Neuroendocrinology Branch
Department of Psychiatry
National Institute of Mental Health
University of Freiburg
National Institutes of Health
Freiburg, West Germany
Bethesda, Maryland
Elizabeth Webster
Robert Stephens
Graduate Student
Assistant Professor
Department of Neuroscience
The Ohio State University
University of Florida School of Medicine
Department of Physiology
Gainesville, Florida
College of Medicine
Columbus, Ohio
Peter J. Whitehouse
Director
Larry W. Swanson
Alzheimer Center
Senior Member
Department of Neurology
Neural System Laboratory
Case Western Reserve University
The Salk Institute
Cleveland, Ohio
La Jolla, California
W. Scott Young, 111
Senior Staff Fellow
Laboratory of Cell Biology
National Institute of Mental Health
Bethesda, Maryland
TABLE OF CONTENTS
Chapter 1
Cloning and Distribution of Expression of the Rat Corticotropin-Releasing
Factor (CRF) Gene.. .....................................................................l
Robert C. Thompson, Audrey F. Seasholtz, James 0. Douglass,
and Edward Herbert
Chapter 2
Distribution and Regulation of Corticotropin-Releasing Factor mRNA in
Brain Using In Situ Hybridization Histochemistry .......................................1 3
W. Scott Young, I11
Chapter 3
Central Nervous System CRF in Stress: Radioimmunoassay Studies ....................2 1
Garth Bissette
Chapter 4
Organization of CRF Imrnunoreactive Cells and Fibers in the Rat Brain:
Irnmunohistochemical Studies ...........................................................2 9
P. E. Sawchenko and L. W. Swanson
Chapter 5
The Organization and Possible Function of Amygdaloid Corticotropin-Releasing
Factor Pathways.. .....................................................................-.5 3
Thackery S. Gray
Chapter 6
Corticotropin-Releasing Factor (CRF) Receptors in the Rat Central Nervous
System: Autoradiographic Localization Studies.. ........................................6 9
Errol B. De Souza and Thomas R. Insel
Chapter 7
Brain Corticotropin-Releasing Factor and Development ................................. 91
Thomas R. Insel
Chapter 8
Neurotransmitter Regulation of the CRF Secretion In Vitro ............................ 107
Michael J. Owens and Charles B. Nemeroff
Chapter 9
Corticotropin-Releasing Factor (CRF) Receptors in Brain: Characterization
and Regulation Studies ................................................................. 115
Errol B. De Souza and Dimitri E. Grigoriadis
Chapter 10
Characterization of Second Messengers Coupled to Corticotropin-Releasing
Factor (CRF) Receptors in Brain.. .................................................... .l37
George Battaglia, Elizabeth L. Webster, and Errol B. De Souza
Chapter 11
Receptors for Corticotropin-Releasing Factor.. ........................................1. 53
Greti Aguilera, Mirza Flores, Pilar Carvallo, James P. Harwood,
Monica Millan, and Kevin J. Catt
Chapter 12
Regulation of Adrenocorticotropic Hormone (ACTH) Secretion by
Corticotropin-Releasing Factor (CRF) .................................................. 175
Catherine Rivier, Mark Smith, and Wylie Vale
Chapter 13
Corticotropin-Releasing Factor (CRF) Actions on Adenohypophyseal
Hormone Secretion by Direct Effects on the Pituitary and by Indirect
Effects on the Hypothalamus ...........................................................l 91
S. M. McCann
Chapter 14
Electrophysiology of Corticotropin-Releasing Factor in Nervous Tissue.. ............., 205
George Robert Siggins
Chapter 15
Effects of CRF on Spontaneous and Sensory-Evoked Activity of Locus
Coeruleus Neurons ....................................................................2. 1 7
Rita J. Valentino
Chapter 16
CRF Effects on EEG Activity: Implications for the Modulation of Normal
and Abnormal Brain States.. ...........................................................2 33
Cindy L. Ehlers
Chapter 17
Behavioral Effects of Corticotropin-Releasing Factor. ..................................2 53
George F. Koob and Karen T. Britton
Chapter 18
Corticotropin-Releasing Factor and Ingestive Behaviors ................................2 67
John E. Morley and Allen S. Levine
Chapter 19
Behavioral and Endocrine Studies of Corticotropin-Releasing Hormone
in Primates ............................................................................2.7 5
Ned H. Kalin
Chapter 20
Regulation of the Autonomic Nervous System by Corticotropin-Releasing Factor.. .... 291
Marvin R. Brown and Laurel A. Fisher
Chapter 21
CRF: Central Nervous System Action to Influence Gastrointestinal Function and
Role in the Gastrointestinal Response to Stress.. ....................................... 299
Yvette Tache, Mark M. Gunion, and Robert Stephens
