General Neurochemical Techniques PDF
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NEUROMETHODS 0 1 General Neurochemical Techniques NEUROMETHODS Program Editors: Alan A. Boulton and Glen B. Baker Series I: IYeurochemistry 1. General Neurochemical Techniques Edited by Alan A. Boulton and Glen B. Baker, 1985 2. Amines and Their Metabolites Edited by Alan A. Boulton, Glen B. Baker, and Judith M. Baker, 1985 3. Amino Acids Edited by Alan A. Boo/ton, Glen B. Baker, and James D. Wood, 1985 4. Receptor Binding Edlted by Alan A. Boulton, Glen B. Baker, and Pave1 Hrdina, 1986 5. Enzymes Edited by Alan A. Boulton, Glen B. Baker, and Peter H. Yu, 1986 NEUROMETH6DS Series I: Neurochemistry Program Editors: Alan A. Boulton and Glen B. Baker NEUROMETHODS 0 1 General Neurochemical Techniques Edited by Alan A. Boulton and Glen B. Baker Humana Press Clifton, New Jersey l Library of Congress Cataloging-in-Pubhcatton Data Main entry under title: General neurochermcal techniques. (Neuromethods ; 1. Series I, Neurochemistry) Includes brblrographies and index. 1. Neurochemistry--Technrque. 2. Neurophystology-- Technique. I. Boulton, A. A. (Alan A.) II. Baker, GlenB., 1947- . III. Series. Neuromethods ; 1. IV. Series: Neuromethods Series I, Neurochemistry. [PNLM: 1. Neurochemistry--methods. Wl NE337G v. 1 / WL 104 G326] QP356.3.G46 1986 591.1’88 85-30503 ISBN 0-89603-07.5-X 0 1985 The Humana Press Inc Crescent Manor PO Box 2148 Clifton, NJ 07015 All rrghts reserved No part of this book may be reproduced, stored m a retrieval system, or transmitted m any form or by any means, electronrc, mechanical, photocopymg, mrcrofrlmmg, recordmg, or otherwise without written permrssron from the Pubhsher Printed in the United States of America Foreword Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology em- ployed by neuroscientists, and those that are available often seem either out of date or limited in scope. This series IS about the methods most widely used by modern-day neuroscientists and is written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routmely across the widest range of sub- disciplines. Collecting these general techniques together in a sin- gle volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 de- scribe current procedures for the analyses of amines and their me- tabolites and of amino acids, respectively. These collections will clearly be of value to all neuroscientists working in or contemplat- ing research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are pro- vided for many types of ligand-receptor binding, including chap- ters on general principles, drug discovery and development, and a most useful appendix on computer programs for Scatchard, nonlinear and competitive displacement analyses. Volume 5 pro- vides procedures for the assessment of enzymes involved in biogenic amine synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neuro-chemists, -pharmacologists, -physiologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical. V 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 searching both of memory and hbrary shelves did it seem that perhaps the publisher was right. Although some excellent methods books have recently appeared, notably in neuroana- tomy, it is a fact that there IS a dearth in this particular field, a fact attested to by the alacrity and enthusiasm with which most of the contributors to this series accepted our invitations and suggested additional topics and areas. After a somewhat hesrtant start, es- sentially in the neurochemistry section, the series has grown and will encompass neurochemistry, neuropsychratry, neurology, neuropathology, neurogenetics, neuroethology, molecular neu- robiology, animal models of nervous drsease, and no doubt many more “neuros. ” Although we have tried to include adequate methodological detail and m 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 limitations we have invrted a guest editor to loin with us on most volumes in order to ensure complete coverage of the field and to add their specialized knowledge and competencies. We anticipate that this series will fill a gap; we can only hope that it will be filled appropriately 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 VI Preface to Volume 1 When I began neurochemical research m Saskatoon after working for several years m the UK on the chemistry of yeast protoplasts and the levels of amme metabolrtes in the body fluids of mentally disordered in- dividuals, I remember wondering, which animal should I select, how do I get its brain out, how do I dissect that brain, how do I slice it or obtain subcellular fractions? Later my worries continued, albeit at a rather more specialized level, such as how do I isolate discrete nuclei, perfuse parts of the brain, or lesion or stimulate it? It seemed to me at the time that a book that described general techniques would have been invaluable. For the first volume m this new series on Neuromethods, therefore, we have tried to assemble the techniques most of us use routinely In Chapter 1, Palkovits describes the microdissection of fresh, fro- zen, or fixed (stained) brains yielding as little as 10 kg wet wt. of a specific nucleus from a variety of species. de Lores Arnaiz and Pellegrino de Iraldi in Chapter 2 describe how to break down cellular integrity and isolate synaptosomes, synaptic vesicles, synaptosomal membranes, mye- lm, Golgi structures, lysosomes, nuclei, mitochondria, microtubules, and filaments. In Chapter 3 Lipton describes the uses and abuses of the brain slice, paying particular attention to its selection, preparation, morphol- ogy, Integrity, mcubation, metabolism, stimulation, and advantages m release studies Hertz et al. m their contribution describe the advantages of using tissue culture and cultured cells from the cerebral cortex and cerebellum, and describe the isolation of astrocytes and oligodendro- cytes and their use m biochemical, pharmacological, and physiological experiments Mishra et al. m their chapter describe how molecular biol- ogy and m particular hybridoma technology can be applied to the prepa- ration of monoclonal antibodies for application in the neurosciences, particularly in the study of neurotransmitter and neuropeptide recep- tors. In more physiological terms, MacDonald (Chapter 6) outlmes the use of microiontophoresis and micropressure techniques m the identifi- cation of central transmitters, describing in some detail the types of re- lease, origms of variability, artifacts, errors, and extra- and mtracellular recording; Greenshaw (Chapter 7) describes electrical and chemical ways to stimulate the various parts and pathways of the brain, paying considerable attention to stereotaxic techniques, anesthesia, electrodes, stimulation circuits, wave forms, cannulae, pumps, micromlection, and Preface damage to injection sites, as well, of course, as several examples of working applications. Pittman et al. in Chapter 8 offer a critical review of perfusion tech- nology; they cover the perfusion medium itself, the extent of tissue dam- age, and the collection of the perfusate, as well as applications (ventricu- lar, spinal cord, in vitro, cup, and local &sue). The basic concepts and methods used to study brain electrical activity are described m Chapter 9 by Vanderwolf and Leung. In particular they discuss the problems of assessing electrophysiological correlates of behavior, before proceeding to describe recording methods, slow waves, evoked potentials, retlculo- cortical and neocortical activity, and the effects of drugs on these active- ties in relation to ammal behavior. After discussing early ablation techniques for creatmg lesions, Schal- lert and Wilcox (Chapter 10) move to a detailed discussion of transmltter- selective lesions created by neurotoxins such as 6-hydroxydopamine, 5,7-dihydroxytryptamine, and the aziridinmm ion of ethylcholme, as well as mentioning some of the newer toxins such as N-2-chloroethyl-N-ethyl- bromobenzylamme and l-methyl-4-phenyl-1,2,3,6-tetrahydropyndine, and their likely future uses. In Chapter 11 Smith provides a critique of earlier single-pass tech- niques for determining blood-bram barrier permeabilmes, and after thorough kmetic analyses concludes that intravenous admuustratron and zn sztu brain perfusion techniques are the most versatile and sensi- tive for measurmg transport of substances mto the bram. In Chapter 12, Brady reviews types of axonal transport, some theo- ries to explain the phenomenon, and the six discrete rate components that exist before describmg in detail the three procedures (direct visuah- zation, labeling, and extrinsic markers) for its assessment Postmortem analyses, collection of human postmortem material in a brain bank, dis- section, and safety, as well as some of the variables that affect neuro- chemical values m postmortem materials, are discussed m the penulti- mate chapter by Reynolds. The effects of age, cause of death, mental disorder, drug treatment, time of death, and time after death, as well as some methods, are all covered Fmally, in the last chapter Sarnat de- scribes the evolution of the vertebrate brain, the relation of evolution to disease, and the application of histochemistry m comparative neuroana- tomy. I hope that this general techmques volume will prove to be as useful today as its counterpart would have been to me 20 years ago Alan A. Boulton Contributors L. BAUCE Department of Medical Physiology, The University of l Calgary, Calgary, Alberta, Canada A.J. BLUME . Rocke Institute of Molecular Biology, Nutley, New Jersey SCOTTT . BRADY 9 Department of Cell Biology, University of Texas Health Science Center at Dallas, Dallas, Texas J. DISTURNAL Department of Medical Physiology, The University l of Calgary, Calgary, Alberta, Canada A.J. GREENSHAW 9 Psychiatric Research Division, University of Saskatchewan, Sash toon, Sash tckewan, Canada L. HERTZ Department of Pharmacology, University of Saskatcke- l wan, Saskatoon, Saskatchewan, Canada B.H.J. JUURLINK Department of Anatomy, University of Saskat- l chewan, Saskatoon, Saskatchewan, Canada L.-W.5 LEUNG Department of Psychology, University of West- l ern Ontarzo, London, Ontario, Canada PETERLIPTON Department of Physiology, University of Wisson- l srn, Madzson, Wisconsin J.F. MACDONALD Department of Pharmacology, Unzverslty of l Toron to, Toronto, On tarzo R. K. MISHRA Departments of Psychiatry and Neurosciences, l McMaster University, Hamilton, Ontario, Canada K.MULLIKIN-KILPATRICK Rocke Institute of Molecular Biology, l Nutley, New Jersey MIKL~S PALKOVITS First Department of Anatomy, Semmelweis l Unzversity Medical School, Budapest, Hungary A. PELLEGRINODE~RALDI lnstituto de Biologia Celular, Buenos l Aires, Argentina Q. J. PITTMAN Department of Medrcal Physiology, The Unzver- l srty of Calgary, Calgary, Alberta, Canada GAVINP.REYNOLDS + Department of Pathology, Queen’s Medical Centre, Unrversity of Nottingham, Nottingham, UK C.IWHAGEN Department of Medical Physiology, The University l of Calgary, Calgay, Alberta, Canada GEORGINA RODRIGUEZDE LORES ARNAIZ Facultad deMedi- l cina, Institute de Biologia Celular and Facultad de Farmacla y Bioquztnica, Universldad de Buenos Aires, Buenos Aires, Argentina HARVEY~ARNAT 9 Vepatments OJP edlatrzcs,P athology, and Clm- zeal Neurosclences, University of Calga y, Calga y, Alberta, Canada TIMOTHY~CHALLERT Department of Psychology and Pharmacol- l ogy, University of Texas at Austin, Austin, Texas QUENTIN R. SMITH Laboratory of Neurosctences, National Znsti- l tute on Aging, Natzonal Institutes of Health, Bethesda, Mayland K. SONNENFIELD Roche Institute of Molecular Bzology, Nut&, l New Jersey S. SZUCHET Department of Neurology, Unzverszty of Chicago, l Chicago, Illinois C. H. VANDERWOLF Department of Psychology, Unrverslty of l Western Ontarzo, London, Ontarzo, Canada W.L. VEALE Department of Medtcal Physzology, The University l of Calgay, Calgary, Alberta, Canada W.WALZ Department of Physiology, University of Saskatchewan, l Saskatoon, Saskatchewan, Canada RICHARD E. WILCOX Department of Pharmacology, Unrverszty l of Texas at Austin, Austin, Texas
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