Front cover

Neurobiology of Steroids

copyright Page

Table of Contents

Contributors to Volume 22

Preface

Methods in Neurosciences

Section I: Steroid Kinetics and Metabolism

Chapter [1]. Steroid Hormone Transport through Blood-Brain Barrier: Methods and Concepts

Introduction

Blood-Brain Barrier Transport: Methodological Approaches

Major Determinants of Blood-Brain Barrier Transport of Steroid Hormones

Steady State Model of Steroid Hormone Distribution in Brain

Concluding Remarks

Acknowledgment

References

Chapter [2]. Enhanced Delivery of Steroids to the Brain Using a Redox-Based Chemical Delivery System

Introduction

Redox Methods for Brain-Enhanced Delivery of Steroids

Synthesis of Brain-Enhanced Estradiol-Chemical Delivery System

Distribution of Estradiol-Chemical Delivery System

Pharmacology

Concluding Remarks

Acknowledgments

References

Chapter [3]. Neurosteroids: Biosynthesis and Function

Neurosteroids: A Brief History

Neurosteroid Metabolism in Brain

Mechanisms of Neurosteroid Action: Genomic Effects

Nongenomic, Membrane Receptor-Mediated Activities of Neurosteroids

Neurosteroid Physiology

Concluding Remarks

Acknowledgments

References

Chapter [4]. Cytochrome P450 Enzymes in Brain

Introduction

Quantitation and Subcellular Distribution of Brain P450

Detection of Specific Forms of P450 in Brain by Catalytic Activities

Arachidonic Acid Metabolism

Immunological Detection of Brain P450

Brain P450 Purification and Characterization

Detection of P450 mRNAS in Brain

Induction of Brain P450

Human Brain P450

General Conclusions

Acknowledgments

References

Chapter [5]. Methods for Estimating 11/β-Hydroxysteroid Dehydrogenase Activity

Introduction

Methods of Detecting 11/β -Hydroxysteroid Dehydrogenase Activity

Concluding Remarks

References

Section II: Steroid Receptors

Chapter [6]. Steroid Hormone Binding to Intracellular Receptors: In Vitro and in Vivo Studies

Introduction

Cell Nuclear Retention of Corticosteroids

Binding Assay for Cytosolic Corticosteroid Receptors

References

Chapter [7]. Steroid Hormone Binding to Membrane Receptors

Introduction

Criteria for Physiologically Relevant Receptors

Methods

Characterization of [3H]Corticosterone Binding to Amphibian Brain Membranes

Concluding Remarks

Acknowledgments

References

Chapter [8]. In Vitro Autoradiography for Steroid Receptors

Introduction

Estrogen Receptors

Corticosteroid, Androgen, and Progestin Receptors

Conclusions and Prospectus

Acknowledgments

References

Chapter [9]. Immunocytochemical Studies on Glucocorticoid Receptor

Monoclonal Antibodies Demonstrate Glucocorticoid Receptor in Rat Central Nervous System

Subcellular Distribution of Glucocorticoid Receptors

Glucocorticoid Receptors and Pain Mechanisms

Conclusions

Acknowledgments

References

Chapter [10]. Confocal Scanning Laser Microscopy of Steroid Receptors in Brain

Introduction

Confocal Scanning Laser Microscopy

Immunolabeling for Confocal Scanning Laser Microscopy: Special Requirements

Confocal Scanning Laser Microscopy of Steroid Receptors in Cultured Cells

Confocal Scanning Laser Microscopy of Steroid Receptors in Brain Tissue

Concluding Remarks

References

[11] Ultrastructural Aspects of Steroid Receptor Localization: Immunocytochemical Perspective

Introduction

Preexperimental Considerations

Ultrastructural Detection of Glucocorticoid Receptor Immunoreactivity in Neural Tissue

Ultrastructure of Estrogen Receptor-Immunoreactive Neural Elements

Concluding Remarks

Acknowledgments

References

Chapter [12]. Hybridization Studies of Adrenocorticosteroid Receptors in the Central Nervous System

Introduction

Northern Blot Hybridization

In Situ Hybridization

Solution Hybridization: Nuclease Protection Assay

Concluding Remarks

Acknowledgments

References

Chapter [13]. Detection and Characterization of Epalon Receptors: Novel Recognition Sites for Neuroactive Steroids That Modulate the GABAA Receptor Complex

GABAA Receptor Complex Active Neurosteroids

Methods of Detection of GABAA Receptor Complex-Active Epalons

Concluding Remarks

Acknowledgments

References

Chapter [14]. Mutation Analysis of Steroid Hormone Receptors

Introduction

Mutational Analysis of Steroid Hormone Receptors

Concluding Remarks

References

Section III: Molecular Effects of Steroids

Chapter [15]. Protein-DNA-Binding Assay for Analysis of Steroid-Sensitive Neurons in Mammalian Brain

Introduction

Methodology for Brain Tissue

Methodological Observations

Concluding Remarks

Acknowledgments

References

Chapter [16]. Gene Transfection Studies Using Recombinant Steroid Receptors

Introduction

Transfection Strategies

Protocols for Transfections and Enzymatic Assays

Practical Approaches to Assay Gene Regulation by Nuclear Hormone Receptors

Concluding Remarks

Acknowledgments

References

Chapter [17]. Regulation of Neuropeptide Genes:Determination of Responsiveness to Steroids and Identification of Receptors in Brain Nuclei

Introduction

Determination of Steroid Responsiveness of Genes by Transient Transfection in Cell Lines

Simultaneous Determination of Nuclear Hormone Receptor mRNA in Microdissected Brain Tissue

Concluding Remarks

References

Chapter [18]. Cloning of Steroid-Responsive mRNAs by Differential Hybridization

Introduction

In Vivo Glucocorticoid Treatment

Poly(A)-Containing RNA Isolation

Selection of XgtlO c\ Insertion Vector

Construction of Hippocampal cDNA Library

Screening cDNA Library by Differential Hybridization

Analysis of Cloned Corticosterone Responses

Concluding Remarks

Acknowledgments

References

Chapter [19]. Molecular Correlates of Corticosterone Action in Hippocampal Subregions

Introduction

Methods

Results

Discussion

Acknowledgments

References

Chapter [20]. Glucocorticoid-Determined Protein Synthesis

Introduction

Methodology

Concluding Remarks

Acknowledgments

References

Chapter [21]. Use of Antisense Oligodeoxynucleotides to Block Gene Expression in Central Nervous System

Introduction

Designing Antisense Oligonucleotide

Delivery of Antisense Oligonucleotides

Controls and Nonspecific Effects

Concluding Remarks

References

Section IV: Cellular Efifects of Steroids

Chapter [22]. Steroid Regulation of Neuronotrophic Activity: Primary Microcultures of Midbrain Raphe and Hippocampus

Introduction

Direct Effect of Steroids on Serotonergic Neurons

Steroid Action on Hippocampal Conditioned Medium

Steroid Action on Hippocampal Neurons

Concluding Remarks

References

Chapter [23]. Central Glucocorticoid Receptors and Neuronal Plasticity

Introduction

Glucocorticoid Receptors and Growth Factors of Neuronal and Glial Origin

Central Glucocorticoid Receptors and Prenatal Development

Central Glucocorticoid Receptors and Postnatal Development

Central Glucocorticoid Receptors and Aging Process

Summary

Acknowledgments

References

Chapter [24]. Steroid Action on Neuronal Structure

Introduction

Golgi Impregnation

Concluding Remarks

Acknowledgments

References

Chapter [25]. Electron Microscopie Double and Triple Labeling Immunocytochemistry in Elucidation of Synaptological Interactions between Ovarian Steroid-Sensitive Neuronsand Circuits

Introduction

Tissue Preparation

Double Immunostaining Techniques

Double Immunostaining Combined with Degeneration

Interconnections between Hypothalamic Steroid-Sensitive Circuits and LHRH Neurons

Concluding Remarks

Acknowledgment

References

Chapter [26]. Gene-Mediated Steroid Control of Neuronal Activity

Introduction

Electrophysiological Techniques

Timing Protocol

Application of Steroids

Future Developments

Concluding Remarks

References

Chapter [27]. Electrophysiological Studies of Neurosteroid Modulation of γ-Aminobutyric Acid Type A Receptor

Introduction

Choice of Preparation and Recording Technique

Cell Culture Techniques

Whole Cell and Single-Channel Recording: Practical Aspects

Applications of Techniques in Investigating Steroid Modulation of GABAA Receptors

Concluding Remarks

Acknowledgment

References

Chapter [28]. In Vitro Approaches to Studying Glucocorticoid Effects on Gene Expressionin Neurons and Glia

Introduction

Immunocytochemical Localization of Glucocorticoid and Mineralocorticoid Receptors in Hippocampal Neurons and Cerebral Astrocytes

Expression of Glucocorticoid and Mineralocorticoid Receptor mRNAs in Cultured Neurons and Astrocytes

Effects of Glucocorticoids on Gene Expression, Using Two-Dimensional Gel Electrophoresis

Applications of in Vitro Systems

Procedures

Concluding Remarks

Acknowledgment

References

Section V: Steroid Effects on Integrated Systems

Chapter [29]. Behavioral Approaches to Study Function of Corticosteroids in Brain

Introduction

Automated Computerized Registration of Behavior

General Preconsiderations for Behavioral Experiments on Effects of Corticosteroids

Corticosteroid Receptors, Information Processing, and Spatial Behavior

Concluding Remarks

Acknowledgment

References

Chapter [30]. Adrenocorticosteroids and Cardiovascular Regulation: Methods for Surgery and Blood Pressure Measurements

Introduction

Rat Models of Mineralocorticoid Hypertension

Preparing Lateral Ventricular Cannulas and Catheters

Infusion Pumps and Solutions

Anesthesia and Surgery

Lateral Ventricular Cannulation

Nephrectomies, Adrenalectomies, and Adrenal Enucleations

Choice of Method to Measure Blood Pressure

Indirect Tail Plethysmography

Concluding Remarks

References

Chapter [31]. Steroids and Central Regulationof Immune Response

Introduction

Control of Cytokine Gene Expression by Glucocorticoids

Concluding Remarks

References

Chapter [32]. Steroid Hormone Effects on Brain: Novel Insights Connecting Cellular and Molecular Features of Brain Cells to Behavior

Introduction

Gene Expression in Brain in Light of Composite Response Elements

Genomic and Nongenomic Effects of Steroids on Brain and Other Tissues

Neurosteroids: Brain as a Source of Steroids

Steroid-Carrier Proteins and Blood-Brain Barrier

Importance of Steroid Metabolism and Compartmentation

Developmental Actions of Steroids

Structural Changes in Brain Not Confined to Development

Conclusions: Connecting Cellular and Molecular Aspects of Steroid Hormone Action to Behavior

Acknowledgments

References

Index

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