NEUROMETHODS n 33

Cell Neurobiology Techniques

NEUROMETHODS Series Editors: A/an A. Boulton and Glen B. Baker

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In Vitro Neurochemical Techniques, edlted by Alan A. Boulton, Glen 6. Baker, and Alan N. Bateson, 1999

Cell Neurobiology Techni ues, edlted by Alan A. Bodton, Glen B. s Ba er, and Alan N. Bateson, 1999

In Vivo Neuromethods, edlted by A/an A. Boufton, Glen B. Baker, and Alan N. Bateson, 1998

C Protein Methods and Protocols: Role of G Protefns m Psychlatfrc and Neufologlcal Dfs- orders, edlted by Ram K. Mishra, Glen B. Baker, and Alan A. Boulton, 1997.

Regulatory Protein Modification: Technques and Protocols, edlted by Hugh C. Hemmings, jr., 1997 Apoptosis Techniques and Protocols, edlted by /udes Poirier, 1997

Transgenic Xenopus: Mlcroqectron Methods and Developmental Neurob,ology, by Shlomo Seidman arid Hermona Soreq, 7997

Voltammetric Methods in Brain Systems, ed- lted by Alan A. Boulton, Glen B. Baker, and Ralph N. Adams, 1995 Pa&h-Clamp Applications and Protocols, ed- lted by Alan A. Boulton, G/en B. Baker, and Wo/(&ng Walz, 1995

Neurotrophic Factors, edlted by A/an A. Boulton, G/en B. Baker, and Franz Hefti, 1993

Animal Models of Drug Addiction, edlted by A/an A. Boulton, Glen 6. Baker, and Peter wu, 1992

Practical Cell Culture Techniques, edited by Alan A. Boulton, Glen B. Baker, and WolfRanR Walz, 1992

Animil Models of Neurological Disease, IkMeta- bolrc Encephlopathres and the Eprlepsres, edtted by Alan A. Boulton, Glen 8. Baker, and Roger F. Buttenvorfh, 1992 Animal Models of Neurological Disease, I: Neurodegenerahve Daease, edlted by Alan A. Boulton, Glen B. Baker, and Roger F. Butterworth, 1992

Intracellular Messen B$too2 Glen B. Ba a

ers, edited by A/an A. er, and Cohn W. Jay-

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Animal Models in Psychiatry, II,edlted by A/an A. Boulton, Glen B. Baker, and Mathew T Martin-lverson, 1991

Animal Models in Psychiatry, I, edrted by A/an A. Boulton, Glen B. Baker, andMathew T. Martin-lverson, 199 1

Neuropsychology, edited by Alan A. Boulton, Glen B. Baker, and Merrill Hiscock, 1990

Molecular Neurobiologlcal Techmques, edrted by Alan A. Boulton, Glen B. Baker, and Anthony T. Campagnoru, 1990

Neurophyslological Technlques:App/xarrons to Neural Systems, edited by Alan A Boulton, Glen B. Baker, and Case H. Vanderwolf, 1990

Neurophysiological Techniques: Basic Meth- ods and Concepts, edited by A/an A. Boulton, Glen B. Baker, and Case H. Vanderwolf, 1990

Psychopharmacology, edtted by Alan A. Boulton, Glen B. Baker, and Andrew 1. Greenshaw, 1989

Drugs as Tools in Neurotransmitter Research, edlted by Alan A. Boulton, Glen B. Baker, and August0 V. luorio, 1989

Carbohydrates and Energy Metabolism, ed- ited by Alan A. Boulton, Glen B. Baker, and Roger F. Butterworth, 1989

Analysis of Psychiatric Drugs, edited by Alan A. Boulton, Glen B. Baker, and Ronald T Coutts, 1988

The Neuronal Microenvironment, edited by Alan A. Boulton, Glen B. Baker, and Wolfgang Wa/z, 1988

Imaging and Correlative Physicochemical Techniques, edited by Alan A. Boulton, Glen B. Baker, and Donald P. Boisvert, 1988

lipids and Related Compounds, edited by A/an A. Boulton, Glen 6. Baker, and Lloyd A. Horrocks, 1988

Peptides, edited by A/an A. Boulton, G/en B. Baker, and Quentin Pittman, 1987

Neurotransmitter Enzymes, edited by A/an A. Boulton, Glen 6. Baker, and Peter H. Vu, 1986

NEUROMETHODS n 33

Cell Neurobiology Techniques

Edited by

Alan A. Boulton Unlverslty of Saskatchewan, Saskatoon, Canada

Glen B. Baker Un/vers/ty of AIberta, Edmonton, Canada

Alan N. Bateson University of Alberta, Edmonton, Canada

H UMANA PRESS TOTOWA, NEW JERSEY

0 1999 Humana Press Inc 999 Rrvervrew Drove, Smte 208 Totowa, New Jersey 07512

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ISBN o-89603-510-7 ISSN 0893-2336

Preface to the Series When the President of Humana Press first suggested that a series

on methods in the neurosciences might be useful, one of us (AAB) was quite skeptical; only after discussions with GBB and some search- ing both of memory and library shelves did it seem that perhaps the publisher was right. Although some excellent methods books had re- cently appeared, notably in neuroanatomy, it was a fact that there was a dearth in this particular field, a fact attested to by the alacrity and enthusiasm with which most of the contributors to this series ac- cepted our invitations and suggested additional topics and areas. After a somewhat hesrtant start, essentially in the neurochemistry section, the series has grown and will encompass neurochemistry, neuropsy- chiatry, neurology, neuropathology, neurogenetics, neuroethology, molecular neurobiology, animal models of nervous disease, and no doubt many more “neuros.” Although we have tried to include adequate methodologrcal detail and in many cases detailed protocols, we have also tried to include wherever possible a short introductory review of the methods and/or related substances, comparisons with other methods, and the relationship of the substances being analyzed to neurological and psychiatric disorders. Recognizing our own limita- tions, we have invited a guest editor to join with us on most volumes in order to ensure complete coverage of the field. These editors will add their specialized knowledge and competencies. We anticipate that this series will fill a gap; we can only hope that it will be filled appro- priately and with the right amount of expertise with respect to each method, substance or group of substances, and area treated.

Alan A. Boulton Glen B. Baker

V

Preface Cell Neurobiology Techmques is one of three volumes that represent an

update of the original Volume 1 of Neuvomethods, General Neurochemical Techniques. The expansion to three volumes was necessitated by the rap- idly increasing number of techniques being used by neuroscientists and the general tendency of neuroscience research to become more multidisciplinary and collaborative in recent years. The two companion volumes are con- cerned with molecular and with whole animal approaches to neurobio- logical research.

Even with three volumes, the coverage of General Neuromethods is obviously not exhaustive. We have attempted to cover those techniques that seem particularly popular in multidisciplinary neuroscience research Though some important techniques may have been omitted, we believe that most of these have been covered in detail in other recent volumes of Neuromethods or will be the subjects of upcoming volumes.

The editors anticipate that this volume devoted to cell neurobiology techniques will be unusally useful to neuroscientists across a wide range of fields-e.g., neurochemistry, neuropsychopharmacology, neurology, psychiatry, and neuroanatomy. Each chapter has been written by well- known neuroscientists with active research programs. Experienced re- searchers in the specific areas treated will appreciate the present updates on those topics, and neuroscientists expecting their research programs to become more multidisciplinary or to expand into new areas of research should find these volumes of enormous value, particularly those sec- tions offering protocols with useful hints about the advantages and po- tential pitfalls of particular techniques.

In the first chapter of Cell Neurobiology Technzques, T. A. Reader and C. Strazielle discuss the use of quantitative autoradiography to study monoamine uptake sites and receptors in brain. Cell cultures have been used widely in neuroscience research, and, B. H. J. Juurlink and W. Walz describe detailed protocols for establishment of neural cell cultures. Cell adhesion molecules have also been of great interest to neuroscientists for many years and recently there has been increased interest in their possible involvement in various cancers and psychiatric disorders; K. Tieu and I’. Yu have written a comprehensive chapter on the study of such molecules. The chapter on the measurement of enzymes and their inhibitors by A. Holt is designed to provide neuroscientists who

VII

. . . V//l Preface

are occasional users of enzymes with a basic understanding of how enzymes function as well as guidance in assay design, data analysis, and avoidance of problems. Drug metabolism techniques are fre- quently employed in studies on mechanisms of drug action and on pharmacokinetic drug-drug interactions, and two chapters related to drug metabolism are included m this volume, in one R. T Coutts, J Fang, M. Bourin, and G. 8. Baker discuss prmcrples of drug me- tabolism, with an emphasis on psychiatric drugs, and in the other chapter P. Glue and R. P. Clement provide a comprehensive chapter on cytochrome P-450 enzymes, which are involved m the metabolism of many drugs. In recent years, there has been a flurry of Interest m the role of immediate early genes (IEGs) in neuronal activity and in the actions of various drugs on the expression of these IEGs T. L. Krukoff has provided an immunohistochemistry chapter on the use of c-j& expression as a marker of functional activity in the brain, and G S. Robertson has written an overview on the use of several IEGs as activity markers in the central nervous system. Over the past 25 years, flow cytometry has been utilized widely in the fields of hematology, rmmunology, oncology, and microbiology, recent evidence suggests that it may also be a useful technique for studying developmental neurobiology, and m their chapter, D. Marie, I. Marie, and J L Barker demonstrate several possible applications of flow cytometry in stud- ies of central nervous system development. The study of human post- mortem brain tissue has done much to advance our knowledge of the etiology of neurologic and psychiatric disorders, and G. I’. Reynolds and C. L. Beasley have written a chapter that provides an overview of the use, value, and potential of studies utilizing human brain tis- sue taken at autopsy. In recent years, great strides have been made m applications of magnetic resonance sprectroscopy (MRS) to human brain metabolism, and C. C. Hanstock and P S Allen have provided a chapter on this exciting area of research

Alan A. Boulton Glen B. Baker

Alan IV. Bateson

Contents

Preface to the Series . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ZI

Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vzi

Contributors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

Quantitative Autoradiography of Monoamine Uptake Sites and Receptors m Rat and Mouse Brain

Tomas A Reader and Catherine Strazielle . .*

Neural Cell Culture Techniques Bernhard H J Juurlznk and Wolfgang Walz.

Neural Cell Adhesion Molecules Kim Tteu and Peter H Yu

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On the Measurement of Enzymes and their Inhibitors Andrew Holt 132

Cytochrome P-450 Enzymes: In Vztro Assessment and Clinzcal lmpltcattons

Paul Glue and Robert P Clement 195

c-fos Expression as a Marker of Functional Activity in the Brain. Immunohtstochemtstry

Teresa L Krukof . . . . . .

Immediate-Early Genes as Activity Markers in the CNS George S Robertson . . . . . . . * *

Principles of Drug Metabolism, with an Emphasis on Psychiatric Drugs

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. ..* 231

Ronald T Courts, Jtan Fang, Mtchel Bourin, and Glen B Baker > 255

Flow Cytometric Strategies to Study CNS Development Dragan Marie, lrina Martc, and Jejj?ery L. Barker ,

Neurochemistry of Human Postmortem Brain Gavm P Reynolds and Clare L. Beasley . . . . . . . . . . .

Applications of Proton MRS to Study Human Brain Metabolism Chrrstopher C Hanstock and Peter S Allen , *. *.. . . . .

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Index .,.,...,.,......,., ,,.,. . . . . . . . . . . . . . . . . .,.....,.,.,.,...,.... .,..,. 382

IX

Contributors PETER S. ALLEN l Department ofBiomedical Engineering, University

@Alberta, Edmonton, Alberta, Canada GLEN B. BAKER l Neurochemical Research Unit, Department of

Psychiatry, Walker Mackenzie Centre, University ofAlberta, Edmonton, Alberta, Canada

JEFFERY L. BARKER l Laboratory of Neurophysiology, NZNDS, NIH, Bethesda, MD

CLARE L. BEASLEY . Department of Biomedical Sciences, The University of Shefield, Sheffield, UK

MICHEL BOIJRIN l Laboratozre de Pharmcologie, Fact&& de Medecine, Universite’ de Nantes, Nantes, France, and Neurochemical Research Unit, Department oj=Psychiatry, Walker Mackenzze Centre, University ofAlberta, Edmonton, Alberta, Canada

ROBERT I?. CLEMENT l Department of Drug Metabolism, Schering Plough Research Instttute, Kenilworth, NJ

RONALD T. COUTTS l Neurochemzcal Research Unit, Department 4 Psychiatry, Walker Mackenzie Centre, University @Alberta, Edmonton, Alberta, Canada

JIAN FANG l College @Pharmacy and Nutrition, Universtty qf Saskatchewan, Saska toon, Saskatchewan, Canada

PAUL GLUE l Department of Clinical Pharmacology, Schering Plough Research Institute, Kenilworth, NJ

CHRISTOPHER C. HANSTOCK l Department sf Biomedical Engmeering, University of Alberta, Edmonton, Alberta, Canada

ANDREW HOLT l CV Technologzes, Edmonton, Alberta, Canada BERNHARD H. J. JUURLINK l Department ofAnatomy and Cell Biology and

The Cameco Multiple Sclerosis and Neuroscience Research Centre, College @Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

TERESA L. KRUKOFF l Department @Anatomy and Cell Biology, Faculty ofMedicine, Untversity ofAlberta, Edmonton, Alberta, Canada

DRAGAN MARIC l Laboratory qf Neurophysiology, NINDS, NZH, Bethesda, MD

IRINA MARIC l Laboratory ofNeurophysiology, NINDS, NH, Bethesda, MD

XI

Xii Contrrbutors

TOMAS A. READER l De’partement de Physiologic, Facul td de Me’decine, Universite’ de Mont&al, Montrkal, Que’bec, Canada

GAVIN I?. REYNOLDS l Department qfBiomedica1 Sciences, The University ofShqj?eld, Shefield, UK

GEORGE S. ROBERTSON l Department oj=Pharmacology, Faculty of Mediczne, University @Ottawa, Ottawa, Ontarzo, Canada

CATHERINE STRAZIELLE l Laboratozre de Neuroanatomie Fonctionelle, Facultt de Chirurgie Detaire, Universzte’ de Henri-Pozncar&Nancy 1, Nancy, France

KIM TIEU l Neuropsychiatrtc Research Unit, Unzverszty of Saskatchewan, Saskatoon, Saskatchewan, Canada

WOLFGANG WALZ l Department of Physiology, College @Medicine, University $Saskatchewan, Saskatoon, Saskatchewan, Canada

PETER H. Yu l Neuropsychiatric Research Unit, University 4 Saskatchewan, Saskatoon, Saskatchewan, Canada