Neurotrophic Factors

Neurotrophic Factors

First edition

Sandra E. Loughlin

James H. Fallon

Department of Anatomy and Neurobiology, College of Medicine, University of California, Irvine, Irvine, California

Academic Press, Inc.

Harcourt Brace Jovanovich, Publishers

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Table of Contents

Cover image

Title page

Copyright page

Contributors

Preface

1: Functional Implications of the Anatomical Localization of Neurotrophic Factors

I INTRODUCTION

II NEUROTROPHIC FACTORS INCLUDED IN THIS VOLUME

III REVIEW OF CELLULAR EVENTS OF NEUROTROPHIC FACTOR SYSTEMS

IV MODES OF SECRETION AND TRANSPORT OF NEUROTROPHIC FACTORS

V NEUROTROPHIC FACTOR DISTRIBUTION IN CENTRAL NEURAL SYSTEMS

VI SUMMARY

ACKNOWLEDGMENTS

2: Neurotrophic Factors: What Are They and What Are They Doing?

I HISTORY

II DEFINITION

III PHYSIOLOGICAL FUNCTIONS OF NEUROTROPHIC FACTORS

IV PHARMACOLOGICAL EXPLOITATION OF NEUROTROPHIC FACTORS

V CONCLUSIONS

3: Synergy, Retrograde Transport, and Cell Death

I INTRODUCTION

II CELL DEATH

III AXOPLASMIC TRANSPORT

IV INTERACTIONS BETWEEN NEURONALLY ACTIVE MOLECULES

V MECHANISMS OF INTERACTION

VI OTHER MECHANISMS TO ACHIEVE A RETROGRADE MESSAGE

VII CONCLUSIONS

4: Primary Response Gene Expression in the Nervous System

I INTRODUCTION

II fos AND jun GENES AS PROTOTYPICAL PRIMARY RESPONSE GENES

III OTHER PRIMARY RESPONSE GENES

IV IN VITRO EXPRESSION OF PRIMARY RESPONSE GENES IN NEURAL SYSTEMS

V IN VIVO EXPRESSION OF PRIMARY RESPONSE GENES

VI SECONDARY RESPONSE GENES AS TARGETS OF PRIMARY RESPONSE GENES

VII SPECIFICITY OF PRIMARY RESPONSE GENE EXPRESSION

5: Nerve Growth Factor and Related Substances: Structure and Mechanism of Action

I INTRODUCTION

II NGF STRUCTURE

III RECEPTORS

IV MECHANISM OF ACTION OF NGF

V SUMMARY AND CONCLUSIONS

ACKNOWLEDGMENTS

6: Regulation of Nerve Growth Factor Expression

I EXPRESSION IN THE TARGET

II EXPRESSION IN NEURAL SUPPORTING CELLS AFTER INJURY

III EXPRESSION IN EARLY DEVELOPMENT AND NONNEURAL SETTINGS

IV BIOLOGIC SIGNIFICANCE OF NGF REGULATION:STRATEGIES FOR MANIPULATION IN VIVO

7: Nerve Growth Factor: Actions in the Peripheral and Central Nervous Systems

I INTRODUCTION

II NGF ACTIONS IN THE PERIPHERAL NERVOUS SYSTEM

III NGF ROLE IN THE CENTRAL NERVOUS SYSTEM

8: Brain-Derived Neurotrophic Factor: An NGF-Related Neurotrophin

I INTRODUCTION

II NGF: PROTOTYPICAL NEUROTROPHIC FACTOR

III SEARCH FOR OTHER NEUROTROPHIC FACTORS

VI DISCOVERY OF NEUROTROPHIC ACTIVITIES DISTINCT FROM NGF

V PURIFICATION OF BDNF FROM PORCINE BRAIN

VI NEURONAL SPECIFICITY OF BDNF

VII BDNF PREVENTS NEURONAL DEATH IN VIVO

VIII MOLECULAR CLONING OF PORCINE, MOUSE, RAT, AND HUMAN BDNF

IX CONSEQUENCES OF THE MOLECULAR CLONING OF BDNF

X BINDING OF BDNF TO SPECIFIC RECEPTORS

XI NEUROTROPHIN FAMILY OF NGF-RELATED NEUROTROPHIC FACTORS—NGF, BDNF, NT-3, AND OTHERS

XII CLINICAL PERSPECTIVE

XIII CONCLUSIONS

ACKNOWLEDGMENTS

9: Biochemistry and Molecular Biology of Fibroblast Growth Factors

I INTRODUCTION

II STRUCTURE AND EXPRESSION OF FGF GENES

III PROTEIN STRUCTURES

IV RECEPTORS

V INHIBITORS

VI SIGNAL TRANSDUCTION

VII BIOLOGIC FUNCTIONS

10: Fibroblast Growth Factors: Their Roles in the Central and Peripheral Nervous System

I INTRODUCTION: THE FGF GENE FAMILY

II LOCALIZATION OF THE FGF GENE FAMILY IN NEURAL CELLS AND TISSUES

III FGF RECEPTORS ON NEURAL CELLS

IV FGFs AND THEIR EFFECTS ON NEURONS

V FGFs AND THEIR EFFECTS ON GLIAL CELLS

VI FGFs IN THE LESIONED NERVOUS SYSTEM: REGULATION AND EFFECTS

VII FGFs IN NEURAL TUMORS

VIII FGFs IN CELL LINEAGE DECISIONS

IX FGFs AND THEIR INTERACTIONS WITH OTHER GROWTH FACTORS

X CONCLUSIONS

ACKNOWLEDGMENTS

11: Epidermal Growth Factor: Structure, Expression, and Functions in the Central Nervous System

I IDENTIFICATION

II MOLECULAR STRUCTURE AND HOMOLOGIES

III EGF LOCALIZATION IN BRAIN

IV EGF RECEPTOR

V EGF RECEPTOR EXPRESSION IN BRAIN

VI EGF AND NEUROGLIA

VII EGF AND NEURONAL FUNCTION

12: Transforming Growth Factors Alpha and Beta

I INTRODUCTION

II HISTORICAL BACKGROUND

III ASSAYS FOR TGFα

IV STRUCTURE AND PROPERTIES OF TGFα

V MOLECULAR BIOLOGY AND GENETICS OF TGFα

VI EFFECTS OF TGFα IN VIVO AND IN VITRO

VII CLINICAL IMPLICATIONS

VIII TGFα AND NEUROBIOLOGY

IX ASSAYS FOR TGFβ

X STRUCTURE AND PROPERTIES OF TGFβ

XI MOLECULAR BIOLOGY AND GENETICS OF TGFβ

XII EFFECTS OF TGFβ IN VITRO AND IN VIVO

XIII TGFβ AND OTHER GROWTH FACTORS

XIV CLINICAL IMPLICATIONS OF TGFβ

XV TGFβ AND NEUROBIOLOGY

XVI CONCLUSIONS AND FUTURE ASPECTS

13: Insulin-Like Growth Factors in the Brain

I INTRODUCTION

II INSULIN-LIKE GROWTH FACTORS IN THE BRAIN

III IGF-BINDING PROTEINS IN THE BRAIN

IV INSULIN AND IGF RECEPTORS

V POSTRECEPTOR MECHANISMS

VI BIOLOGIC ACTION OF INSULIN AND IGFS

VII HYPOTHESIS

ACKNOWLEDGMENTS

14: Neurobiology of Insulin and Insulin-Like Growth Factors

I INTRODUCTION

II NEURAL CIRCUITRY AND INSULIN-LIKE GROWTH FACTORS

III IGFs AND INSULIN VIEWED WITHIN THE CONTEXT OF NEUROTROPHIC THEORY

IV SHARED BIOCHEMICAL PATHWAY FOR NEURITE OUTGROWTH

V OTHER NEUROBIOLOGICAL ROLES

VI PATHOPHYSIOLOGY

ACKNOWLEDGMENTS

15: Ciliary Neuronotrophic Factor

I HISTORICAL BACKGROUND : 1940–1975

II FROM CILIARY NEURON FACTOR CONCEPT TO FIRST IDENTIFICATION: 1976–1980

III MOLECULAR STUDIES LEADING TO MAMMALIAN CNTF PURIFICATION: 1981–1990, AND GENE CLONING: 1989–1991

IV BIOLOGIC PROPERTIES

V SUMMARY AND PERSPECTIVES

ACKNOWLEDGMENTS

16: Skeletal Muscle-Derived Neurotrophic Factors and Motoneuron Development

I IN VITRO EVIDENCE FOR ACTIONS OF SKELETAL MUSCLE NEUROTROPHIC FACTORS

II IN VIVO EVIDENCE FOR ACTIONS OF SKELETAL MUSCLE NEUROTROPHIC FACTORS

III ISOLATION AND IDENTIFICATION OF SKELETAL MUSCLE-DERIVED TROPHIC FACTORS

IV EFFECTS OF PURIFIED FACTORS ON MOTONEURON SURVIVAL IN VIVO

ACKNOWLEDGMENTS

17: Growth Factors for Myelinating Glial Cells in the Central and Peripheral Nervous Systems

I INTRODUCTION

II CENTRAL NERVOUS SYSTEM GLIA

III PERIPHERAL NERVOUS SYSTEM GLIA

IV CONCLUSIONS AND OUTSTANDING QUESTIONS

ACKNOWLEDGMENTS

18: Adhesion Factors

I INTRODUCTION

II LAMININ AND FIBRONECTIN

III POTENTIATION OF NEUROTROPHIC ACTIVITY OF PEPTIDE GROWTH FACTORS BY LAMININ AND FIBRONECTIN

IV PROTEOGLYCANS

V ACTIVE COMPLEXES OF LAMININ AND FIBRONECTIN WITH PROTEOGLYCANS

VI CELL–CELL CONTACT-MEDIATED NEURONAL SURVIVAL

VII PURPURIN AND APOLIPOPROTEINS

VIII GANGLIOSIDES

IX CONCLUSIONS

ACKNOWLEDGMENTS

19: Instructive Neuronal Differentiation Factors

I PHENOTYPIC PLASTICITY

II CHOLINERGIC DIFFERENTIATION FACTOR/LEUKEMIA INHIBITORY FACTOR

III CILIARY NEUROTROPHIC FACTOR

IV MEMBRANE-ASSOCIATED NEUROTRANSMITTER- STIMULATING FACTOR

V SWEAT GLAND FACTORS

VI FACTORS ACTING ON MOTOR NEURONS

VII NORADRENERGIC FACTORS

VIII PEPTIDERGIC FACTORS

IX CORTICOSTEROID

X GONADAL STEROIDS

XI MORPHOLOGICAL FACTORS

XII MELANIZATION FACTORS

XIII NEURONAL ACTIVITY

XIV OTHER FACTORS

XV MUTANTS IN INVERTEBRATES

XVI HEMATOPOIETIC ANALOGY

20: Neurotransmitters as Neurotrophic Factors

I INTRODUCTION

II AMINO ACID TRANSMITTERS

III MONOAMINES

IV NEUROPEPTIDES

V CONCLUSIONS

ACKNOWLEDGMENTS

Index

Copyright

Copyright © 1993 by ACADEMIC PRESS, INC.

All Rights Reserved.

No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage and retrieval system, without permission in writing from the publisher.

Academic Press, Inc.

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United Kingdom Edition published by

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Library of Congress Cataloging-in-Publication Data

Neurotrophic factors / edited by Sandra E. Loughlin and James H. Fallon.

p. cm.

Includes bibliographical references and index.

ISBN 0-12-455830-5

1. Neurotrophic functions. 2. Growth factors. I. Loughlin, Sandra E. II. Fallon, James H.

QP363.N52 1992

591.1’88–dc20

92-11683

CIP

PRINTED IN THE UNITED STATES OF AMERICA

92 93 94 95 96 97 MV 9 8 7 6 5 4 3 2 1

Contributors

Numbers in parentheses indicate the pages on which the authors’ contributions begin.

Martin L. Adamo(391),     Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892

Joseph G. Altin(129),     Division of Cell Biology, John Curtin School of Medical Research, Australian National University, Canberra, ACT, 2601 Australia

Alaric T. Arenander(89),     Department of Anatomy and Cell Biology, The Mental Retardation Research Center, and The Laboratory of Biomedical and Environmental Sciences, UCLA School of Medicine, Los Angeles, California 90024

Carolyn Bondy(391),     Developmental Endocrinology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892

Ralph A. Bradshaw(129),     Department of Biological Chemistry, College of Medicine, University of California, Irvine, Irvine, California 92717

Ellen J. Collarini(489),     Department of Biology, University College of London, London WC1E 6BT, United Kingdom

Michael F. Crouch(51),     Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra ACT 2601, Australia

Timothy L. Denton(25),     Division of Neurogerontology, Andrus Gerontology Center, University of Southern California, Los Angeles, California 90089

Robert H. Edwards(181),     Department of Neurology, UCLA School of Medicine, Los Angeles, California 90024

James H. Fallon(1),     Department of Anatomy and Neurobiology, College of Medicine, University of California, Irvine, Irvine, California 92717

Mark L. Grimes(209),     Department of Neurology, University of California, San Francisco, San Francisco, California 94121

Glaudia Grothe(313),     Department of Anatomy and Cell Biology, University of Marburg, Robert-Koch-Straβe 6, D-3550 Marburg, Germany

Theo Hagg(443),     Department of Biology, University of California, San Diego, La Jolla, California 92093

Franz Hefti(25),     Andrus Gerontology Center, University of Southern California, Los Angeles, California 90089

Ian A. Hendry(51),     Division of Neuroscience, John Curtin School of Medical Research, Australian National University, Canberra ACT 2601, Australia

Harvey R. Herschman(89),     Department of Anatomy and Cell Biology, Department of Biological Chemistry, The Mental Retardation Research Center, and The Laboratory of Biomedical and Environmental Sciences, UCLA School of Medicine, Los Angeles, California 90024

David M. Holtzman(209),     Department of Neurology, University of California, San Francisco, San Francisco, California 94121

Douglas N. Ishii(415),     Departments of Physiology and Biochemistry, Colorado State University, Fort Collins, Colorado 80523

Beat Knusel(25),     Division of Neurogerontology, Andrus Gerontology Center, University of Southern California, Los Angeles, California 90089

Paul A. Lapchak(25),     Division of Neurogerontology, Andrus Gerontology Center, University of Southern California, Los Angeles, California 90089

Derek LeRoith(391),     Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892

Frances M. Leslie(565),     Department of Pharmacology, College of Medicine, University of California, Irvine, Irvine, California 92717

Ronald M. Lindsay(257),     Regeneron Pharmaceuticals Inc., Tarrytown, New York 10591

Frank M. Longo(209),     Department of Neurology, University of California, San Francisco, San Francisco, California 94121

Sandra E. Loughlin(1),     Department of Anatomy and Neurobiology, College of Medicine, University of California, Irvine, Irvine, California 92717

Jean-Claude Louis(443),     Department of Biology, University of California, San Diego, La Jolla, California 92093

Gerson Lüdecke(313),     Department of Anatomy and Cell Biology, University of Marburg, Robert-Koch-Straβe 6, D-3550 Marburg, Germany

Marston Manthorpe(443),     Department of Biology, University of California, San Diego, La Jolla, California 92093

James L. McManaman(475),     ¹ Department of Neurology, Division of Neuroscience and Program in Cellular and Molecular Biology, Baylor College of Medicine, Houston, Texas 77030¹

Present affiliation: Department of Neuroscience, Synergen, Inc., Boulder, Colorado 80301.

William C. Mobley(209),     Departments of Neurology, Pediatrics, and the Neuro-science Program, University of California, San Francisco, San Francisco, California 94121

Richard Morrison(339),     R. S. Dow Neurological Sciences Institute and Comprehensive Cancer Center, Portland, Oregon 97209

Hans W. Müller(509),     Molecular Neurobiology Laboratory, Department of Neurology, University of Düsseldorf, Moorenstr. 5, D-4000 Düsseldorf, Germany

Ronald W. Oppenheim(475),     Department of Neurobiology and Anatomy and Neuroscience Program, The Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina

Dörte Otto(313),     Department of Anatomy and Cell Biology, University of Marburg, Robert-Koch-Straβe 6, D-3550 Marburg, Germany

Paul H. Patterson(527),     Division of Biology, California Institute of Technology, Pasadena, California 91125

Pauli Puolakkainen(359),     Bristol-Myers Squibb Pharmaceutical Research Institute—Seattle, Seattle, Washington 98121

Mohan Raizada(391),     Department of Physiology, University of Florida, Gainesville, Florida 32610

William D. Richardson(489),     Department of Biology, University College of London, London WC1E 6BT, United Kingdom

Charles T. Roberts, Jr(391),     Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892

Kenneth A. Thomas(285),     Department of Biochemistry, Merck Research Laboratories, Rahway, New Jersey 07065

Daniel R. Twardzik(359),     Bristol-Myers Squibb Pharmaceutical Research Institute—Seattle, Seattle, Washington 98121

Klaus Unsicker(313),     Department of Anatomy and Cell Biology, University of Marburg, Robert-Koch-Straβe 6, D-3550 Marburg, Germany

Silvio Varon(443),     Department of Biology, University of California, San Diego, La Jolla, California 92093

Haim Werner(391),     Diabetes Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892

Reiner Westermann(313),     Department of Anatomy and Cell Biology, University of Marburg, Robert-Koch-Straβe 6, D-3550 Marburg, Germany

Preface

Although it has been known since the early 1950s that growth factors affect neural tissues, it is only in the past decade that some characterized growth factors have been shown to be localized in the central nervous system. In the past five years, we have seen an explosion in research into growth factors, their receptors and their functional roles in development, reorganization, and pathology. This is thus an appropriate time to comprehensively review the information on neurotrophic factors in mammalian nervous systems.

This volume provides not only current reviews, but also attempts to provide a conceptual framework for understanding the rich spectrum of actions of neurotrophic factors. The members of the major growth-factor families, including the neurotrophins, the epidermal growth factor family, and the fibroblast growth factors, are comprehensively reviewed, along with a number of more recently discovered factors. Multifaceted approaches to the study of growth-factor effects on specific and defined neuronal groups are discussed. The actions of neurotrophic factors on first, second, and third messenger systems, as well as their convergence onto primary response genes are examined. Synergistic effects and interactions between neurotrophic factors are described, especially as they affect the complex glial–neuronal mixture characteristic of the nervous system. In addition to the traditionally defined neurotrophic factors, the growth-promoting effects of neurotransmitters, neuronal activity, and interactions with the extracellular matrix are considered. Thus, this volume brings together specific reviews of known growth factors and offers a conceptual discussion of their anatomical distributions, modes of actions, and interactions.

The editors gratefully acknowledge the contributions of Nanette Canepa in preparation of the index. Kathleen Cooper of the American Parkinson Disease Association SCC continues to serve as an inspiration to us all.

1

Functional Implications of the Anatomical Localization of Neurotrophic Factors

James H. Fallon; Sandra E. Loughlin

I INTRODUCTION

This volume is devoted to the analysis and review of a class of growth factors that acts on neural tissue, namely, neurotrophic factors. The history of growth factor and neurotrophic factor research can be presented differently, depending on the definitions used and the scientific discipline of the writer. As neuroanatomists, we trace the first conceptualization of neuronal trophic factors to the visionary neuroanatomist, Ramón y Cajal (1882, 1891; De Felipe and Jones, 1991). This concept, together with studies by Hamburger (1934) on neuronal development, had its foundation in a developmental Zeitgeist of 19th century European embryologists (see Chapter 2). Growth factor research per se is believed to have had its experimental genesis in 1916 when Robertson coined the term tethelin for the growth-stimulating activity of pituitary extracts in vitro (reviewed by Burgess, 1989). A decade later, insulin was recognized for its broad growth-promoting effects (Banting and Best, 1922). Growth factors were shown to be present in the brain over 50 years ago, when it was demonstrated that brain homogenates contained factors that stimulated fibroblasts to divide in vitro (Trowell et al., 1939; Hoffman, 1940). This fibroblast growth factor activity later was purified partially and was shown to have effects on many other tissues (Gospodarowicz, 1974). During the 1950s, the landmark studies by Levi-Montalcini and Hamburger (1953) established a conceptual and experimental blueprint for subsequent studies in the field of growth factors in the nervous system.

Despite a wealth of in vitro and in vivo molecular, biochemical, physiological, and pharmacological data generated on growth factors over the past 50 years, relatively little anatomical information on the localization of growth factors has been available until the past decade. This has been especially true for neurotrophic factors in the central nervous system. For example, epidermal growth factor was isolated 30 years ago by Cohen (1962) but the anatomical localization of epidermal growth factor-like immunoreactivity in the brain was not known until the 1980s (Fallon et al., 1984). This lag in morphological information is, in part, due to the low levels of neurotrophic factors present in the brain. However, new sensitive immunocytochemical, in situ hybridization, and receptor autoradiographic techniques have afforded the neuroanatomist powerful tools for locating neurotrophic factor systems in the brain. With these techniques, an abundance of neuroanatomical information has been gathered at the light microscopic level in the past few years that has confirmed predictions by Ramón y Cajal on the presence and importance of trophic factors in the brain.

In this introductory chapter, we will present some neuroanatomical perspectives on the roles of neurotrophic factors in the brain. Because of the sheer volume of the newly available anatomical information on the distribution of neurotrophic factors in the nervous system, it is impractical to present atlas mappings of all known neurotrophic factors. This chapter will stress some key morphological issues concerning neurotrophic factors in the central nervous system and how knowledge of the distribution of these growth factor systems is critical to our understanding of how these substances may be involved in developing, adult, aging, and damaged nervous systems.

II NEUROTROPHIC FACTORS INCLUDED IN THIS VOLUME

Growth factors include substances that stimulate cells to divide (hyperplasia) or increase in size (hypertrophy). Many growth factors are now known to exist (Table 1). Trophic factors include those substances that have effects on cell differentiation, survival, phenotypic expression, and plasticity, as well as on cell hypertrophy, for example, neurite extension (also considered a trophic action). Neurotrophic factors, a subset of growth factors acting on neural tissue, have been defined in a myriad of ways ranging from very restrictive to very general. Restrictive definitions are based on the nerve growth factor model, which recognizes only the specific aspects of developmental events, such as cell survival and neurite extension, that are supported by proteins and peptides. This restrictive definition of a neurotrophic substance parallels the restrictive definition of and requirements for a neurotransmitter, which were based on experimental paradigms developed for acetylcholine. Coincidentally, nerve growth factor and acetylcholine not only have forged our fundamental definitions of a neural growth factor and a neurotransmitter, respectively, but also are associated closely in brain function. As more putative neurotransmitters and neurotrophic factors have been discovered and analyzed, it has become clear that broader definitions of each group are necessary to capture the range of subtle and profound actions of these neuromessengers.

Table 1

Examples of growth factors

A focused definition that integrates both traditional and modern components of neurotrophic activity is presented by Hefti and co-workers in this volume (Chapter 2). This definition states that neurotrophic factors are endogenous, soluble proteins regulating survival, growth, morphological plasticity, or synthesis of proteins for differential functions of neurons. This volume includes chapters on soluble proteins that fulfill these formal criteria, as well as on other protein and nonprotein factors that could be considered neurotrophic factors based on their trophic and trophic-related functions.

The neurotrophic factors (as just defined) discussed in this volume include nerve growth factor (NGF; Chapters 5, 6, and 7), brain-derived neurotrophic factor (BDN; Chapter 8), ciliary neurotrophic factor (CNF; Chapter 15), fibroblast growth factors (FGF; Chapters 9 and 10), insulin and insulin-like growth factors (IGF; Chapters 13 and 14), epidermal growth factor (EGF; Chapter 11), transforming growth factors α and β (TGF; Chapter 12), and skeletal muscle-derived neurotrophic factors (Chapter 16). The effects of a number of growth factors, especially platelet-derived growth factor, on glial cells are also described (Chapter 17). In addition to the roles of these factors, the roles of adhesion factors (Chapter 18), neurotransmitters, and neuropeptides (Chapter 20) as neurotrophic factors are included by virtue of their important neurotrophic activity. These chapters focus on our present knowledge of the neurotrophic factors (first messengers), their receptor systems, and associated transducing mechanisms (second messengers). Many neurotrophic factors effect phenotypic responses in cells through rapid ligand-induced primary response genes (third messengers). These cellular mechanisms are discussed in Chapter 4. Synergistic interactions between neurotrophic factors and intrinsic cellular systems, especially with respect to cell death and survival, are examined in Chapter 3. The role of neurotrophic factors and other cellular mediators as phenotype-specifying factors is developed in Chapter 19. These factors include neurotransmitters, neuropeptides, steroids, and membrane-bound substances, as well as neuronal activity. Thus, links are formed between neurotrophic activity and related cellular mechanisms.

III REVIEW OF CELLULAR EVENTS OF NEUROTROPHIC FACTOR SYSTEMS

Each of the chapters in this volume details specific molecular and cellular aspects of neurotrophic factors. In this section, a brief overview of these events is summarized (see Fig. 1). Although the major cellular events of trophic factor functions are similar for all tissue types, the highly specialized structure and function of central neural tissue presents unique problems for neurotrophic factor transport, availability, synergy, and regulation. The two major cell types in central neural tissue are neurons and glia. In Fig. 1, neuron A is shown receiving an axonal terminal from neuron B. Neuron B, in turn, projects to neuron C. A glial cell is shown sending out processes that communicate with neuron A and with a blood vessel. The nucleus of neuron A contains DNA and the cytoplasm of neuron A is shown to contain rough endoplasmic reticulum (8), Golgi apparatus (10), and polyribosomes (9). Retrograde and anterograde transport are highlighted by arrows outside the axon on neuron A.

Figure 1 ). Vesicles are indicated by circles (O). Abbreviations: BV, blood vessel; ret, retrograde transport; ant, anterograde transport; ecm, extracellular matrix; poly, polyribosomes. See text for discussion.

In Fig. 1, neuron A is depicted containing membrane receptors for neurotrophic factors (1). After binding of the factor to its receptor, signal transduction proceeds through second messenger systems that may alter a number of events through control of gene expression (see, for example, Chapter 5). These events include alterations in synthesis of neuromodulators and growth factors, regulation of differentiation and survival, inhibition or facilitation of inherent death programs (Chapter 3), modulation of morphological plasticity, and regulation of retrograde transport through stabilization of tubulin mRNA (Chapter 14). Neurotrophic factor signal transduction may also regulate membrane (ion fluxes), cytosolic, and cytoskeletal (2) function. The effects of neurotrophic factors on cytoskeletal elements actually may be detrimental to the recovery of damaged neural systems. For example, Butcher and Woolf (1989) have proposed that, in some cases (e.g., Alzheimer’s disease), neurotrophic factors may precipitate a pathologic cytoskeletal cascade that accelerates neuronal degeneration. Regulation and buffering of ion concentration may be an important function of neurotrophic factors. The control of Ca²+ levels through modulation of calcium-binding proteins (calmodulin, 28k calbindin, parvalbumin), enzymes (protein kinase C, mitochondrial systems, Ca²+ ATPase), and membrane Ca²+ channels may be how neurotrophic factors afford protection against excitotoxins.

Interactions between neurotrophic factors and other neurotrophic agents (3,4) are known to occur (Chapter 3). Some neurotransmitters and neuropeptides that mediate neurotransmission at synapses possess neurotrophic activity (4) (Chapter 20). Synergy between neurotrophic factors, and between neurotrophic factors and neurotransmitters with neurotrophic activity, may be mediated through convergence on second and third messenger systems, where interactions can occur at the level of the second messengers or the phosphorylation of substrates (Chapter 4). Convergence of neurotrophic factor cascades also occurs at the level of the third messenger systems (primary response gene products), as indicated in Fig. 1 (5,6,7). Thus, a number of neurotrophic factors are able to stimulate primary response genes such as c-fos and c-jun (5). Primary response genes are transcribed to mRNAs that are translated to proteins, for example, FOS and JUN (6,8), which then activate target genes (7), leading to further cell-specific synthesis of new proteins on polyribosomes (9) or rough endoplasmic reticulum (8).

Neurotrophic factors that contain consensus secretory signal sequences for insertion through membranes (e.g., EGF, TGFα, TGFβ, NGF) are synthesized in the rough endoplasmic reticulum (8); further posttranslational modifications are carried out in the vesicular system of the Golgi apparatus (10), from which the factors may be transported and released from vesicles (11,12,13). Neurotrophic factors lacking a consensus secretory signal sequence (e.g., aFGF, bFGF, CNTF, IL-ls) are thought to be synthesized in polyribosomes and released into the cytosol (Janet et al., 1987; Chapters 9 and 10). The absence of secretory leader sequences and glycosylation in these neurotrophic factors poses an obvious problem. How are they released from cells? Holocrine secretion after cell death and lysis (see Fig. 2) is one probable mode of secretion. Other physiological modes of release are possible, but as yet undefined. There are, interestingly, many other examples of proteins of this type. For example, annexin 1, which also contains no signal secretory sequence, is nevertheless secreted by the merocrine portion of the prostate gland (Christmas et al., 1991).

Figure 2 Modes of secretion and intercellular communication mechanisms of neurotrophic activity. Abbreviations: g, glial; n, neuronal; bv, blood vessel; ecm, extracellular matrix.

Neurotrophic factors synthesized in neurons may be secreted at a number of sites, for example, cell somata (12), dendrites (not illustrated), axons, and axon terminals (13). The problem of long distance availability of trophic factor action is obviated in neural tissue by the elaboration of the axon and its numerous terminal branches. For example, dopamine neurons of the substantia nigra of the rat project axons 10 mm or more in length to the caudate putamen (nigrostriatal pathway), where each axon may give rise to 500,000 terminals (Anden et al., 1966). In addition, a single dopamine neuron may give rise to axons that collateralize to widely divergent forebrain targets such as caudate putamen, septum, and cortex (Fallon and Loughlin, 1982). Thus, a single neuron could provide neurotrophic factors to many brain regions, as well as retrogradely transport (Fig. 1) neurotrophic factor-receptor complexes (17) or related messengers (18) (Chapters 3 and 14) from many brain regions. Therefore, the elaborate axonal (and dendritic) processes in neural tissue provide a unique morphological substrate for neurotrophic factor access and availability.

Highly branched axonal systems also provide one clue for apparent anatomical mismatches between neurotrophic factors and their receptors. For example, neuron A may project to neuron C. Neuron C releases a growth factor (16) that binds to a receptor on the axon terminal of neuron A (17,18). This case shows no anatomical mismatch between the neurotrophic factor and its receptor. However, if neuron A also projects an axon collateral to another brain site that lacks the neurotrophic factor synthesized by neuron C, the axons and axon terminals of neuron A in this other brain site may still express receptors for the neurotrophic factor secreted by neuron C. This would result in an apparent mismatch between the neurotrophic factor and its receptor.

Glia also synthesize and respond to neurotrophic factors (20,21,22). Like neurons, glia display regional heterogeneity (Chapter 19); a subset of astrocytes contain TGFα-precursor immunoreactivity (Fallon et al., 1990). Glia not only synthesize and release neurotrophic factors that may interact with neurons (25), the vasculature (22), and other glia, they also may be key intermediaries of neurotrophic activity between neurons. This sequential signaling may proceed from neuron A to neuron C, through an intermediary glial step (20). Vascular-neuronal trophic interactions may also be mediated by intervening glia. Glucose uptake and extracellular ion concentrations (e.g., Ca²+) are also regulated by glia (21) and may be under neurotrophic factor control.

Neurotrophic factor availability may be restricted or enhanced by adhesion factors, for example, in the extracellular matrix of neural cells and blood vessels (Fig. 1). The extracellular matrix may provide binding or storage sites for active neurotrophic factors (23) or may mask neurotrophic factors in a way that renders them inactive unless there is injury at this site (24). FGFs are sequestered by extracellular matrix heparan proteoglycans that may act as temporary storage sites of FGF (Chapter 4) and participate in the binding of FGF to its receptor (Rapraeger et al., 1991). FGF could be released after anatomical disruption of these sites, leading to neurotrophic and angiogenic effects.

Another potential intercellular communication system for neurotrophic factors may be mediated by juxtacrine interactions (Ankelesaria et al., 1990; Chapter 12) (25 in Fig. 1; see also Fig. 2). In this model of interaction, the neurotrophic factor remains inserted in the membrane of the cell that synthesizes the factor (in Fig. 1, the glial cell is the cell synthesizing the neurotrophic factor) but is still able to bind the receptor located in the responsive cell (neuron A, Fig. 1). This hypothesis has been put forward by Wong et al. (1989) and Ankelesaria et al. (1990) for the binding of TGFα to the EGF receptor in adjacent cells in vitro (Chapter 12). Thus, it is possible that neurotrophic factor interactions in the central nervous system are mediated by similar intercellular mechanisms.

Many of the molecular and cellular events relating to trophic factor function have been studied in peripheral tissues and in vitro. Few of these events, however, have been examined for neurotrophic factors in the central nervous system and much of our specific knowledge remains fragmentary. Several questions regarding the functional morphology of neurotrophic factors have been presented in this section, but we are hampered by a lack of ultrastructural data on the localization of neurotrophic factor and their related molecular systems. At present, much of our morphological information is emerging from light microscopic, immunocyto- chemical, in situ hybridization, and receptor autoradiographic studies. These studies offer insight into some basic questions. Are the neurotrophic factors, their receptors, and the synthetic cellular machinery present in neurons or glia? Even this question is often difficult to answer directly with the techniques used in in situ hybridization and receptor autoradiographic techniques. Electron microscopic studies will be necessary to answer this most basic question. Further, the highly differentiated and compartmentalized nature of central neural tissue poses more difficult morphological questions. The dendritic and axonal arborizations of neurons are elaborate and heterogeneous. Neurotrophic factor molecular systems may be active only in restricted segments of the dendrites of a neuron. In order to determine the precise site of action and interaction of a particular neurotrophic factor, it is important to determine the subcellular localization of the various neurotrophic factors, their binding proteins and relevant adhesion factors, and their receptor, second messenger, and third messenger systems. A single neuron, responsive to several neurotrophic factors, may have multiple domains of functional activity depending on the local neural, glial, vascular, and extracellular matrix relationships of portions of its dendritic and axonal processes. Further, the neurotrophic factor effects on morphological plasticity must be studied in vivo by quantitative electron microscopic techniques.

IV MODES OF SECRETION AND TRANSPORT OF NEUROTROPHIC FACTORS

The specialized nature of neural tissue affords a variety of modes of secretion, transport, and availability of neurotrophic factors. As discussed in the previous section, features of neural cells such as extensive axonal arborization of neurons and neuronal–glial–vascular arrangements provide numerous means of long- and short-distance interactions between neurotrophic factor-releasing and -responsive cells. These modes of secretion and transport are summarized in this section (Fig. 2).

The most widely cited mode of neurotrophic factor transport is retrograde transport of the ligand–receptor complex or its second messenger (Retrograde, Fig. 2) (Chapter 3). The classic example of a neurotrophic factor-receptor complex undergoing this mode of transport is NGF. The second mode of transport of neurotrophic factors is anterograde axonal transport (Anterograde, Fig. 2). This mode has been observed for bFGF in the visual pathway from retinal ganglion cells to the lateral geniculate nucleus and superior colliculus (Ferguson et al., 1990).

Neurotrophic factors also are thought to act locally in the central nervous system through several mechanisms, including local actions of soluble factors on adjacent neurons and glia (paracrine, Fig. 2) and the binding of membrane-bound neurotrophic factors and their receptors on adjacent cells (juxtacrine). Local actions also might include the self-regulation of production by the binding of a neurotrophic factor to its receptors in the same cell or by nuclear targeting of a cytosolic neurotrophic factor (autocrine). One hypothetical subset of autocrine regulation is termed intracrine (Chapter 8) and refers to purely intracellular regulatory mechanisms. Recently, Clevenger et al. (1991) have provided evidence that peptide hormones may function in the nucleus without first binding to a cell-surface receptor, lending credence to the concept that some growth factors may function in an intracrine manner. After the death or anatomical disruption of a cell, a neurotrophic factor such as aFGF, which lacks a secretory signal sequence, can be released from the dying cell (holocrine). Factors secreted by a cell may gain access to the extracellular space, through which they diffuse to act at great distances from the secretory cell (telocrine). This mode of action has not been demonstrated convincingly in the central nervous system.

Some modes of transport unique to the central nervous system include movement of neurotrophic factors between the cerebrospinal fluid and the parenchyma of the brain and spinal cord. These one-way and two-way movements of factors occur through ependymal cells and modified ependymal cells called tanacytes (ependymal). Communication between the blood and the central nervous system occurs selectively across blood vessels participating in the blood–brain barrier as well as across circumventricular organs lacking this barrier. Release of neurotrophic factors into the blood in these regions would constitute a neuroendocrine mode of secretion.

Selective transport between the blood and the central nervous system is regulated by endothelial cells of blood vessels and transport mechanisms of astrocytes (vascular). Both glial and neuronal interactions with blood vessels are restricted, in part, by the basal laminae and associated extracellular matrix elements. The movement of insulin and insulin-like growth factors across the blood–brain barrier and through circumventricular organs lacking a blood–brain barrier is discussed by Le Roith and colleagues (Chapter 13).

Finally, an important mode of neurotrophic factor transport is through a combination of neuronal–glial interactions (sequential). For example, neurotrophic factors derived from one neuronal population first may act locally on glia that, in turn, release a second neurotrophic factor that regulates functions in a separate population of neurons. This possibility is raised by a number of experimental findings. For example, EGF and bFGF increase dopamine uptake in substantia nigra dopamine neurons in vitro only when glia are present (Knusel et al., 1990). This issue is discussed further in a subsequent section of this chapter.

V NEUROTROPHIC FACTOR DISTRIBUTION IN CENTRAL NEURAL SYSTEMS

A Approaches in Neuroanatomy

The molecular and cellular distribution of neurotrophic factor systems was discussed in Section III. In this section, issues relating to the broader distribution of neurotrophic factors in the central nervous system will be discussed. One approach used in neuroanatomical research involves a regional analysis of the central nervous system, which considers areas of the brain and spinal cord with respect to common location, structure, development, and patterns of connectivity (e.g., thalamus, cortex). A second approach is the study of neural systems, which examines the neuraxis with respect to a common functional system (e.g., visual system, extrapyramidal motor system, limbic system). A third parallel approach, chemical neuroanatomy, examines the central nervous system with respect to a particular neurochemical (e.g., acetylcholine, dopamine, enkephalin, neurotrophic factors). All three approaches have provided important perspectives on the functions of neurotrophic factors in the central nervous system.

B Regional Analysis

The hippocampus is a brain region that has received a great amount of attention in neuroanatomical studies of neurotrophic factors. The hippocampus is a phylogen- etically ancient form of cortex (archicortex) with a relatively simple laminated structure. Its cell types and their connections have been studied in great detail for over 50 years (Lorente de Nó, 1934). The hippocampus has received particular attention in neurotrophic factor research because it has been known to be a major source of nerve growth factor for cholinergic neurons of the nucleus basalis of Meynert, which degenerate in patients with Alzheimer’s disease (Wilcock et al., 1983; Whitehouse et al., 1987).

The most popular working hypothesis for the past decade has been that nerve growth factor is synthesized and released by cells in the hippocampus, binds to a nerve growth factor receptor on terminals of cholinergic neurons, and is retrogradely transported as a ligand–receptor complex to cell bodies in the nucleus basalis, where a second messenger signal mediates further trophic activity. Several findings from various disciplines have challenged, or at least broadened, this model of the central neural growth factor systems. These new findings are discussed in the chapters in this volume, but some of the recent neuroanatomical information is discussed here.

First, nerve growth factor is present in other cholinoceptive regions of the brain (Shelton and Reichardt, 1986) and some cholinergic brain areas do not respond to NGF (Knusel and Hefti, 1988). Second, cholinergic neurons are positioned to be under the influence of nerve growth factor that is synthesized in the local environs of their cell bodies (Lauterbom et al., 1991) as well as at distant sites such as the hippocampus. Third, some cholinoceptive brain regions do not contain nerve growth factor but may contain a separate neurotrophic factor instead. Fourth, a cholinoceptive region such as the hippocampus contains not only nerve growth factor, but also other members of the neurotrophin family (BDNF, NT-3) as well as members of other families of neurotrophic factors. This point is illustrated in Fig. 3. These illustrations are derived from in situ hybridization and/or immunocyto-chemical studies on the localization of several neurotrophic factor systems, including NGF (Ernfors et al., 1990; Maisonpierre et al., 1990; Senut et al., 1990); BDNF (Ernfors et al., 1990; Hofer et al., 1990; Maisonpierre et al., 1990; Phillips et al., 1990; Wetmore et al., 1990), neurotrophin-3 (NT-3) (Ernfors et al., 1990; Maisonpierre et al., 1990; Phillips et al., 1990), acidic FGF (aFGF) (Fallon et al., 1991), basic FGF (bFGF) (Pettman et al., 1986; Wilcox and Unnerstall, 1991), IGF-I (Noguchi et al., 1987), and TGFα precursor (Fallon et al., 1990; J. H. Fallon and S. E. Loughlin, unpublished observations).

Figure 3 Coronal sections of hippocampus of the right side of the rat brain illustrating the distribution of neurotrophic factors in this structure. All distributions are in neurons, with the exception of TGFα-precursor, which is localized in glia. The top left panel indicates regions of the hippocampus, including the dentate gyrus (DG) with its molecular (M), granule cell (G), and hilar (H or CA4) layers. Ammon’s horn includes the CA1, CA2, and CA3 regions, each with an alveus (A), stratum oriens (O), pyramidal layer (P), stratum radiatum (R), and stratum lacunosum (L). The fasciola cinerium (FC) is also illustrated.

As illustrated in Fig. 1, NGF (mRNA and peptide) is localized in scattered pyramidal and nonpyramidal neurons throughout the dentate gyrus, hilus, and Ammon’s horn (fields CA1, CA2, CA3). The second member of the neurotrophic family, BDNF, is localized in the same areas as NGF, but a higher proportion of dentate granule cells and Ammon’s horn pyramidal neurons exhibits BDNF mRNA expression. The third member of the neurotrophic family, NT-3, has a more restricted distribution, primarily in dentate granule cells and pyramidal neurons of CA2 and medial CA1. Other levels of complexity of these distributions are present. For example, NT-3 mRNA is highly expressed early in development, BDNF mRNA is expressed later in development after neurogenesis is complete, and NGF mRNA is expressed more consistently throughout development (Maisonpierre et al., 1990). The heterogeneity of receptor subsystems for the neurotrophins adds to the complexities of this neurotrophic system (Chapters 5 and 8).

The hippocampus also contains other neurotrophic activity. For example, bFGF is localized in pyramidal neurons of CA2 and FC (fasciola cinerium). aFGF is sparse, scattered, and found mostly in nonpyramidal neurons. IGF-I is localized in a moderate number of dentate granule cells and in Ammon’s horn pyramidal neurons in CA1, CA2, and CA3. In contrast to these neurotrophic factors, TGFα-precursor is present in a subpopulation of astrocytes throughout the hippocampus.

The plots in Fig. 3 illustrate the rich and heterogeneous distribution of neurotrophic factors in neurons and glia of the hippocampus. Such a regional analysis of neurotrophic factor distribution in the central nervous system highlights the rich diversity of potential growth factor interactions in one brain region and helps isolate specific subregions that may be particularly susceptible to, or resistant to, certain pathological processes. For example, the CA2 region is endowed with a variety of neurotrophic factors (Fig. 3). The CA2 region is particularly resistant to pathological changes in temporal lobe epilepsy, perhaps because of the protective effects of 28K calbindin, which is enriched in this region of the hippocampus (Sloviter et al., 1991), and because of the extensive presence of neurotrophic factors in these neurons. Other protective mechanisms are undoubtedly involved. For example, the presence of glutamate dehydrogenase in astrocytes in these hippocampal regions (Aoki et al., 1987) may offer pyramidal neurons a measure of resistance to gluta- minergic neurotoxicity. Knowledge of the neuroanatomical distribution of neurotrophic factors and regulatory enzyme systems is essential to our understanding of pathological processes that tend to occur (or not occur) in specific brain sites.

C Systems Analysis

1 Extrapyramidal motor system

One central neural system that has received considerable attention in the area of neurotrophic factor research is the extrapyramidal motor system. This system includes a broad spectrum of forebrain, brainstem, and spinal cord structures that participate in the temporal and spatial regulation of the three great motor systems of the body: the somatic (striated voluntary muscles), the limbic/autonomic (involuntary muscles of the gut, cardiovascular system, glands), and the endocrine (pituitary regulated) (Fallon and Loughlin, 1987). These monumental tasks are undertaken by a significant mass of the central nervous system, especially the cortex, basal ganglia, and limbic system. Therefore, we will focus on select subsystems. The subsystems, however, comprise key elements in our study of the etiology of and treatments for Parkinson’s disease and Huntington’s disease.

Fig. 4 outlines some fundamental connections between a few regions involved in the extrapyramidal motor system. Projections from pyramidal neurons in the deep layers of cortex reach the caudate putamen (one sector of the striatum), which then projects to the globus pallidus (one sector of the pallidum) and substantia nigra (another sector of the pallidum also containing dendrites of dopaminergic neurons). The globus pallidus projects to the entopeduncular nucleus (or internal segment of the globus pallidus). The entopeduncular nucleus and substantia nigra pars reticulata project, in turn, to thalamic nuclei, which send a return projection to cortex, completing a major cortico-subcortical loop. Descending projections from the substantia nigra pars reticulata to the deep superior colliculus and pe- dunculopontine nucleus ultimately lead to descending projections to motoneurons of the brainstem and spinal cord. These so-called cortico-striato-pallidal output projections are composed of numerous parallel and convergent channels that contain topographically as well as nontopographically organized systems that program motor behavior (for review, see Fallon and Loughlin, 1987). One pathway that appears to facilitate focused throughput in this system is the ascending dopaminergic projection from the substantia nigra to the caudate putamen, that is, the nigrostriatal pathway. The cardinal pathologic finding in Parkinson’s disease is degeneration of this pathway. In Huntington’s disease, on the other hand, the cardinal pathologic finding is the degeneration of striatal neurons that have descending projections to the globus pallidus and substantia nigra. One major thrust in neurotrophic factor research is determining therapeutic strategies to reverse these progressive pathologic changes and to develop appropriate transplantation techniques to replace the degenerated neural systems. Understanding the distribution of neurotrophic factors in these brain regions is one of the critical first steps in developing therapeutic strategies for these extrapyramidal disorders.

Figure 4 Some basic connections of the extrapyramidal motor systems. Top , Parasagittal section of the rat brain showing cortico-subcortical loops, with projections from cortex (c) to caudate putamen (cp, or dorsal striatum), then to globus pallidus (gp), substantia nigra (snr), and entopeduncular nucleus (ep). ep and snr project to thalamus (t), which returns a projection to cortex; snr also projects to the superior colliculus (sc) and pedunculopontine nucleus (ppn), which project to motoneuronal regions (m) of the brainstem and spinal cord. Bottom , Expanded view of some of these connections in the substantia nigra. The dopamine neurons (DA) of the substantia nigra pars compacta (snc) are shown projecting to and receiving projections from the cp. GABAergic neurons of the substantia nigra pars reticulata (snr) are depicted containing aFGF (FGF). cp terminals of striato-nigral axons contain EGF and local glia contain TGFα precursor (TGF). Bottom right , Magnified view of these neurotrophic factors in the snr. See text for discussion.

2 Striatum

Examination of the distributions of neurotrophic factors in the striatal and pallidal/nigral regions reveals some interesting patterns (Fig. 5). First, the caudate putamen, which contains nigral dopaminergic terminals that degenerate in Parkinson’s disease and striatal neurons that degenerate in Huntington’s disease, does not contain an impressive array of known neurotrophic factors (Fig. 5). NGF, NGF mRNA, and NGF low molecular weight receptor mRNA are found in the striatum (Shelton and Reichardt, 1986; Mobley et al., 1989; Senut et al., 1990), which may serve to act neurotrophically for local cholinergic interneurons, but these levels are also quite low. There is a report of the presence of a striatal-derived neuronotrophic factor (Dal Toso et al., 1988) and IGF-I (Noguchi et al., 1987) in the striatum, but the combined levels of known neurotrophic factors appear to be rather low in the striatum, especially in comparison with other extrapyramidal brain areas. Further, it is not clear how noncholinergic striatal neurons, which constitute over 95% of striatal neurons, are influenced by neurotrophic factors. For example, anterograde transport to, or retrograde axonal transport of neurotrophic messengers from, other extrapyramidal structures may provide access to neurotrophic factors. In contrast, a dramatic increase of TGFα-precursor immunoreactivity in astrocytes in the transplant site is seen following fetal midbrain suspension transplants into an adult dopamine-denervated striatum (Loughlin et al., 1989). Intrastriatal infusions of TGFα, which decrease lesion-induced behavioral deficits, also increase TGFα,(S.E. Loughlin and J. H. Fallon, unpublished observations). These findings suggest that TGFα, which binds to the EGF receptor, may be an important neurotrophic factor in Parkinson’s disease therapy.

Figure 5 Parasagittal sections of brain illustrating localization of EGF in axons in pallidal brain regions ( top ), TGFα precursor in a subpopulation of astrocytes throughout the brain, but especially in pallidal regions ( middle ), and aFGF in neuronal cell bodies throughout the brain, again concentrated in pallidal regions ( bottom ).

A potentially important manifestation of neurotrophic factor heterogeneity in the striatum relates to the progression of neurodegeneration in Huntington’s disease. This disease presents gradients of degeneration of striatal neurons, that is, there is an early loss of striatal enkephalin projections to the external globus pallidus and a later loss of striatal substance P projections to the internal globus pallidus, with relative sparing of enkephalin projections to the substantia nigra pars compacta (Reiner et al., 1988). These findings suggest differences in the susceptibility of matrix and patch neurons of the striatum, perhaps because of differences in neurotrophic factor availability as well as intrinsic lethal cellular programs.

3 Pallidum/Substantia Nigra

The pallidum consists of the globus pallidus, ventral pallidum, entopeduncular nucleus (internal segment of globus pallidus in primates), and substantia nigra pars reticulata. These structures are separated in the brain by incursions of the internal capsule, cerebral peduncle, and medial forebrain bundle, but they have similar cell types and some common histochemical features. One interesting feature of these pallidal structures is the unique distribution of EGF-like immunoreactivity (Figs. 5 and 6) (Fallon et al., 1984; Alheid and Heimer, 1988) that is present in striatal neuronal processes and terminals (J. H. Fallon and S. E. Loughlin, unpublished observations) in these regions (Fig. 6). Note the highly restrictive distribution of EGF and the complex geometry and physical separation of the pallidal structures in which EGF is localized. Such a distribution makes it difficult for nonneuroanatomical techniques (such as whole brain assays or incomplete dissections) to detect EGF in the brain. In addition to EGF in axons and terminals, neuronal cell bodies in these pallidal structures contain aFGF immunoreactivity (Figs. 5 and 7a) and numerous astrocytes contain TFGα-precursor immunoreactivity (Figs. 5 and 7b) (Fallon et al., 1990, 1988). IGF-I has also been reported to be localized in the globus pallidus (Noguchi et al., 1987). Thus, the pallidal regions appear, at present, to be particularly enriched in a multitude of neurotrophic factors.

Figure 6 EGF-like immunoreactivity (DAB-immunoperoxidase technique) in pallidal regions of the brain. (A) Islands of Calleja sector of ventral pallidum. (B) Globus pallidus and downward extension into the ventral pallidum. (C) Entopeduncular nucleus. (D) Substantia nigra pars reticulata. (Photo micrographs courtesy of J. Fallon and K. Seroogy.)

Figure 7 Neurotrophic factor immunoreactivities (DAB-immunoperoxidase technique) in the substantia nigra pars reticulata. (A) aFGF in neuronal cell bodies in pars reticulata and in a portion of the pars compacta (arrow). (B) TGFα precursor in astrocytes.

One pallidal structure of particular interest in disorders of the extrapyramidal motor systems in the substantia nigra, which is composed of the pallidal-like pars reticulata and the overlying pars compacta, which contains dopaminergic neurons projecting to the forebrain and dendrites that invade the pars reticulata (Fig. 4). What is the neurotrophic milieu of these cells? As discussed earlier, and as illustrated in Fig. 4, EGF-like immunoreactivity is present in striatal fiber and terminals known to project to dopaminergic dendrites and GABAergic neurons in the pars reticulata. Many of the large GABAergic neurons in this area contain aFGF. TGFα-precursor is present in a subpopulation of astrocytes in the pars reticulata. In vitro studies have shown that EGF and bFGF stimulate cell proliferation and dopamine uptake in nigral cultures, but this effect depends on the presence of glia (Knusel et al., 1990). The morphological differentiation of embryonic dopaminergic neurons also depends on the presence of astrocytes (Lieth et al., 1989). The compartment- alization of neurotrophic factors in different neuronal and glial elements in the substantia nigra (Fig. 5) and the receptor binding characteristics of EGF and TGFα to the EGF receptor (Chapters 11 and 12), and aFGF and bFGF to FGF receptors (Chapters 9 and 10) suggest a number of plausible interactions in the substantia nigra. One working hypothesis is that a sequential neuronal–glial interaction (Fig. 2) mediates neurotrophic activity after damage. For example, FGFs (without a consensus secretory signal) are released from nigral neurons on injury. Increases in FGF activity have been reported following lesions in other regions of the central nervous system (Nieto-Sampedro et al., 1988b); bFGF has been shown to be retrogradely transported in the nigrostriatal pathway (Ferguson et al., 1990). In response to injury (and in aged animals), EGF-receptor immunoreactivity appears in reactive astrocytes (Nieto-Sampedro et al., 1988a). FGF receptors are present on neurons (Type 1) and glia (Type 2) (Chapter 10) and FGF binding is known to inhibit EGF binding in other tissues (Chapter 9). Thus, the neurotrophic effects of FGF may be mediated through regulation of astrocytic TGFα synthesis, release, and binding to local EGF (and, perhaps, independent TGFα) receptors on glia and neurons. By inhibiting further EGF binding, FGF may regulate continued neurotrophic effects using autocrine, paracrine, and juxtacrine mechanisms.

Dopaminergic neurons also may be regulated tonically by long-distance axonal transport mechanisms. These mechanisms include anterograde and retrograde transport of neurotrophic factors in any of a number of structures interconnecting with the substantia nigra, for example, anterograde transport of EGF from the striatum. Other factors could be transported from the striatum by anterograde or retrograde mechanisms. A partially characterized (15-kDa) striatal-derived neuronotrophic factor has been reported to be neurotrophic for dopaminergic neurons (Dal Toso et al., 1988). BDNF also has neurotrophic effects on these neurons (Zigmond and Strieker, 1989; Knusel et al., 1990; Hyman et al., 1991) but its source in vivo is still in question, since BDNF mRNA is not detectable in striatum (Maisonpierre et al., 1990). Opioid peptides also are known to possess neurotrophic activity (Chapter 20) and the substantia nigra receives dense dynorphin and enkephalin inputs from the striatum (see Fallon and Ciofi, 1990, for review).

In conclusion, fiber pathways, local neurons, and glia of the substantia nigra contain a mosaic of resident neurotrophic factors. The anatomical localization of these factors provides a substrate for complex neurotrophic activity in the intact and damaged substantia nigra.

VI SUMMARY

The central nervous system possesses the molecular and cellular machinery to synthesize, release, and respond to a wide variety of heterogeneously distributed neurotrophic factors. The unique morphology of neural tissue provides a substrate for long-distance transport of neurotrophic factors, as well as for local interactions between neurons, glia, the vasculature, and cerebrospinal fluid. A regional analysis of the central nervous system reveals that one brain area, for example, the hippocampus, may contain members of several families of neurotrophic factors. The distribution of the neurotrophins (NGF, BDNF, NT-3), the fibroblast growth factors (aFGF, bFGF, and others), IGF-1, and TGFα reveals that both distinct and overlapping neurotrophic activity may occur for local neurons and glia, as well as for input and output systems. A systems analysis of the central nervous system reveals that the extrapyramidal motor system contains system-specific neurotrophic factors such as pallidal EGF, as well as a rich assortment of other neurotrophic factors that are present also in other systems. Different levels of the extrapyramidal motor system have been found to contain a spectrum of known neurotrophic factors. For example, the pallidum is enriched in the neurotrophic factors studied to date, whereas the striatum is relatively lacking in such factors. An analysis of neurotrophic factors in the substantia nigra reveals a differential distribution of EGF, aFGF, and TGFα in axons, cell bodies, and astrocytes. These factors may subserve a sequential type of neurotrophic activity linking neurons and glia in the intact and damaged extrapyramidal motor system.

A major function of the nervous system is to process and store information. Many of the changes associated with such activity are similar to those that occur in ontogeny. Phenomena such as long-term potentiation, learning, and adaptation are associated with measurable changes in metabolic activity, ion fluxes, receptor/transmitter/enzyme expression, synapse formation, spine growth, and glial proliferation. Thus, a continuum of processes can be mediated by many of the same messengers, including neurotransmitters and neurotrophic factors.

ACKNOWLEDGMENTS

This work was supported by NIH grant NS 15321 to J. H. Fallon and N1H grant NS 26761 to S. E. Loughlin, as well as by a Bud Corbin Neuroscience Award and grants from the APDA SCC. We wish to thank Pippa Jones for typing the manuscript.

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