Methods in Neuroendocrinology

253 Pages·1998·56.954 MB·\253

by  Louis D. Van de Kar (Author)

METHOD
in NEUROENDOCRINOLOGY
CRC Press
METHODS IN THE LIFE SCIENCES
Gerald D. Fasman - Advisory Editor
Brandeis University
Series Overview
Methods in Biochemistry
John Hershey
Department of Biological Chemistry
University of California
Cellular and Molecular Neuropharmacology
Joan M. Lakoski
Department of Pharmacology
Penn State University
Research Methods for Inbred Laboratory Mice
John P. Sundberg
The Jackson Laboratory
Bar Harbor, Maine
Methods in Neuroscience
Sidney A. Simon Joseph Af. Corless
Department of Neurobiology Department of Cell Biology,
Duke University Neurobiology and Ophthalmology
Duke University
Methods in Pharmacology
John H. McNeill
Professor and Dean
Faculty of Pharmaceutical Science
The University of British Columbia
Methods in Signal Transduction
Joseph Eichberg, Jr.
Department of Biochemical and Biophysical Sciences
University of Houston
Methods in Toxicology
Edward J. Massaro
Senior Research Scientist
National Health and Environmental Effects Research Laboratory
Research Triangle Park, North Carolina
CELLULAR AND MOLECULAR
NEUROPHARMACOLOGY
Joan M Lakoski, Advisory Editor
The CRC Press Cellular and Molecular Neuropharmacology Series provides
the reader with state-of-the-art research methods that address the cellular
and molecular mechanisms of the neuropharmacology of brain function
in a clear and concise format. Topics covering all aspects of
neuropharmacology are being reviewed for publication.
Published Titles
Molecular Regulation of Arousal States, Ralph Lydic
Neuropharmacology Methods in Epilepsy Research, Steven L. Peterson
and Timothy E. Albertson
Methods in Neuroendocrinology, Louis D. Van de Kar

METHOD
in NEUROENDOCRINOLOGY
Edited by
Louis D. Van de Kar
Department of Pharmacology and Experimental Therapeutics
Loyola University of Chicago
Stritch School of Medicine
Maywood, Illinois
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
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First issued in hardback 2019
© 1998 by Taylor & Francis Group, LLC
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No claim to original U.S. Government works
ISBN-13: 978-0-8493-3363-7 (Pbk)
ISBN-13: 978-1-138-45391-3 (hbk)
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Library of Congress Card Number 98-3241
Library of Congress Cataloging-ln·Publlattlon Data
Methods in neuroendocrinology I [edited by) Louis D. Van de Kar.
p. em.·· (Cellular and molecular neuropharmacology)
Includes bibliographical references and index.
ISBN 0-8493-3363-6 (alk. paper)
I. Neuroendocrinology--Methodology. I. Van de Kar, Louis D.
II. Series: CRC Press methods in the life sciences. Cellular and
molecular neuropharmacology.
[DNLM: 1. Neurosecretory Systems--physiology. 2. Neurosecretion
-physiology. 3. Endocrine GIands--physiology. 4. Hormones-
-physiology. 5. Neuroendocrinology--methods. WL 102 M5934 1998)
QP356.4.M48 1998
612.8--dc21
DNLM/DLC
for Library of Congress 98-3241
CIP
Preface
Neuroendocrinology is a scientific discipline concerned with the inter-relationship
between neuronal and endocrine systems. Thus, neuroendocrine research addresses
the regulation of hormone secretion by the brain as well as the influence of hormones
on brain function. Because neuroendocrinology is transitional between endocrinol­
ogy (the study of hormones) and neuroscience (the study of neuronal systems), it
often is lost between these two disciplines. Many classical endocrinologists view
neuroendocrinology as neuroscience while many neuroscientists view neuroendo­
crinology as endocrinology.
This book is intended for students, pre- and post-doctoral fellows, medical
residents and others who are interested in neuroendocrine research. The presentation
of the chapters is designed to allow the readers to obtain insight into the many
scientific disciplines that comprise neuroendocrine research. Thus, the chapters in
the book will describe experimental approaches that bridge endocrine and neurosci­
entific research.
To help in reading this book, a few basic definitions may help guide the reader.
A hormone is a substance that is released from one organ into the blood stream, it
travels in the blood and then induces a physiological reaction in another organ. An
endocrine gland is the organ that secretes the hormone. Feedback refers to a sequence
of events that begins with the secretion of the hormone and results in the inhibition
(or stimulation) of further secretion of the same hormone.
The hypothalamus is the part of the brain most intimately involved in the
regulation of hormone secretion. It is located at the base of the brain on the midline,
ventral to the thalamus and dorsal to the pituitary gland. The hypothalamus regulates
the pituitary gland. The pituitary gland, in turn, controls the function of several
endocrine glands, such as the adrenal gland, thyroid gland, testes and ovaries. The
pituitary gland is divided into three lobes: the anterior, posterior and intermediate
lobes. The hypothalamus controls the secretion of hormones from the distinct lobes
of the pituitary gland through different mechanisms as detailed below. The anterior
lobe of the pituitary gland secretes the following hormones: luteinizing hormone
(LH), follicle stimulating hormone (FSH), adrenal corticotropic hormone (ACTH),
thyroid stimulating hormone (TSH), growth hormone (GH) and prolactin. Hypotha­
lamic releasing factors that are released from the hypothalamus into the pituitary
portal blood vessels reach the anterior lobe of the pituitary gland and stimulate the
secretion of ACTH, FSH, GH, LH, TSH and prolactin. In addition, the secretion of
some hormones, notably prolactin and GH, is kept under tonic inhibition by hypo­
thalamic inhibiting factors that reach the pituitary via the pituitary portal vessels.
Hormones secreted by the anterior lobe of the pituitary gland can also act as trophic
hormones that regulate the growth and overall function of the target glands.
The posterior lobe of the pituitary gland is also known as the neural lobe because
it is an extension of neurons originating in the hypothalamus. Hypothalamic cells
containing vasopressin and oxytocin send their axons into the posterior pituitary
where they release these hormones from their nerve terminals into the circulation.
The intermediate lobe of the pituitary gland contains high concentrations of various
peptides, including endorphin, melanocyte stimulating hormone (MSH) and ACTH
but their function in humans (if any) is not yet known.
Many brain regions that are involved in regulating emotions (i.e., the limbic
system) and homeostasis have afferent and efferent connections with the hypothal­
amus. These regions include the amygdala, hippocampus, bed nucleus of the stria
terminalis, dorsal raphe nucleus, locus coeruleus, parabrachial nucleus and nucleus
of the solitary tract. Neurons located in these brain regions can convey information
relevant to neuroendocrine control to hypothalamic neurons and receive information
from hypothalamic neurons. The processing and integration of neuronal information
flow relevant to the secretion of hormones is a primary focus of neuroendocrine research.
A major function of the neuroendocrine system is to ensure the survival of the
individual and the species. Situations that endanger individual survival can be exter­
nal in origin, for example, pursuit by a predator or the sudden appearance of the
boss in the office, or conditions due to endogenous changes, for example the chest
pain associated with sudden coronary vascular constriction. Other occurrences, such
as famine, might not present an immediate danger to the individual but may endanger
the survival of the species. Survival of the individual is regulated by different
neuroendocrine response mechanisms than the survival of the species. Hormones
responsible for the survival of the individual are often referred to as “stress hor­
mones”, such as the hypothalamic-pituitary-adrenal axis hormones (ACTH, corti-
costerone/cortisol), the renin-angiotensin system and epinephrine. The survival of
the species is more dependent on hormones that regulate reproduction, such as LH
and FSH. A complex network of afferent inputs from sensory organs, from the
viscera and from other regions in the body convey messages to the brain regarding
changes in the external and/or internal environment. The brain integrates these inputs
and organizes the appropriate endocrine, behavioral and autonomic responses to
these environmental or internal challenges.
This book will delineate some of the many different disciplines that are involved
in investigations of neuroendocrine control mechanisms and will indicate what can
be learned from different approaches to neuroendocrine research. The chapters in
the book also will outline the limits of interpretation for several approaches to
neuroendocrine research. Beginning with the most elementary approaches to study­
ing neuroendocrine regulation, chapters in this book will gradually increase in levels
of complexity. Chapter 1 will deal with cellular and molecular biological methods
in immortalized cell cultures of neurons. Chapters 2 to 4 will present molecular
biological and immunocytochemical methodologies in neuroendocrine research. The
following chapters (5 to 7) will describe more complex neural circuits using in vitro
slice preparations. Chapters 8 to 10 will explain how to study neuroendocrine control
mechanisms and the influence of stress in conscious experimental animals. Analyses
of the pulsatile nature of hormone release, both in experimental animals and in
humans, will be described in Chapter 11. The final chapter will describe the inves­
tigation of neuroendocrine function in humans and how hormone responses to
specific stimuli can be used to obtain information about the functional status of the
brain, for example in patients who suffer from depression and schizophrenia.
It is my hope that the chapters in this book will encourage readers who are not
involved in neuroendocrine research to pursue this exciting area. For those among
the readers who are performing research in neuroendocrinology, the following chap­
ters might provide insight into the value of methodological approaches that are not
currently employed in their laboratories.
Louis D. Van de Kar