Glen B. Baker - Böcker

Many of the trace amines-more correctly called biogenic amines- have been known for decades, but because of their tiny concentra- tions (0. 01-100 ng/g) in brain, it was only after the development of sophisticated analytical techniques (such as mass spectrometry) that they could be identified and quantitated in nervous tissue. There are now more than 20 of them and most are related to the catecholamines and 5-hydroxytryptamine both structurally and metabolically. Their pharmacological and physiological properties make them prime candidates for a transmitter or neuromodulator role and many of them elicit profound behavioral syndromes after injection--one of them, phenylethylamine, has even been referred to as nature's amphetamine. In the clinical sphere several have been shown to be involved in: Parkinsonism, schizophrenia, depression, agoraphobia, aggression, hyperkinesis, migraine, hypertensive crises, hypertyrosinemia, he- patic encephalopathy, epilepsy, and cystic fibrosis.Thus the research reported here on these intriguing "new" substances will be of great interest to psychiatrists, neurologists, biochemists, pharmacologists, physiologists, psychologists, behaviorists and indeed to all those working in the neurosciences and related fields today. ACKNOWLEDGMENTS This book is based on the proceedings of Trace Amines and the Neurosciences, a meeting held at the University of Alberta, Edmonton, July 19-21, 1983. This meeting was organized as a Satellite Meeting of the Ninth Meeting of the International Society for Neurochemistry, held in Vancouver, July 10-15, 1983. International organizers of the satellite meeting were Drs. A. A. Boulton (Saskatoon), W. G. Dewhurst (Edmonton), G. B. Baker (Edmonton), and M. Sandler (London).

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscientists, and those that are available often seem either out of date or limited in scope.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 s- disciplines. Collecting these general techniques together in a s- gle volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 - scribe current procedures for the analyses of amines and their - tabolites and of amino acids, respectively. These collections will clearly be of value to all neuroscientists working in or contempl- ing research in these fields.Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are p- vided for many types of ligand-receptor binding, including ch- 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 p- 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.

Techniques u-r the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, 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 neuroscrentrsts and 1s written by their colleagues who are practicing experts Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the widest range of subdiscrplines Collectmg these general techmques together in a single volume strikes us not only as a service, but will no doubt prove of exceptronal utilrtarian value as well Volumes 2 and 3 - scribe all current procedures for the analyses of ammes and their metabolrtes and of ammo acids, respectively. These collectrons will clearly be of value to all neuroscrentrsts working m or contemplating research in those fields.Similar reasons exist for Volume 4 on receptor bmdmg techniques since experimental - tails are provided for all types of lrgand-receptor binding, including chapters on general principles, drug discovery and - velopment, and a most useful appendix on computer programs for Scatchard, nonlinear, and competitrve displacement analyses. Volume 5 provides procedures for the assessment of enzymes- volved m brogemc amme synthesis and catabolrsm. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physrologrsts, -anatomrsts, psychopharmacologrsts, psychratrrsts, neurologrsts, and chemists (organic, analytrcal, pharmaceutrcal, medicinal), m fact, everyone involved m the neurosciences, both basic and clinical.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neurosclentlsts, and those that are available often seem either out of date or limited m scope. This series is about the methods most widely used by modern-day neurosclentists and 1s wrltten by their colleagues who are practicing experts. Volume 1 will be useful to all neurosclentists since It concerns those procedures used routinely across the widest range of subdisciplines. Collecting these general techniques together m a single volume strikes us not only as a service, but will no doubt prove of exceptIona utilitarian value as well. Volumes 2 and 3 - scribe all current procedures for the analyses of amines and their metabolltes and of ammo aads, respectively. These collections will clearly be of value to all neuroscientlsts working m or contemplating research m these fields.Similar reasons exist for Volume 4 on receptor binding techniques since experimental - tails are provided for all types of llgand-receptor bmdmg, including chapters on general prmciples, drug discovery and - velopment, and a most useful appendix on computer programs for Scatchard, nonlinear, and competitive displacement analyses. Volume 5 provides procedures for the assessment of enzymes - volved m biogenic amme synthesis and catabolism. Volumes in the NELJROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomlsts, psychopharmacologists, psychlatnsts, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal), in fact, everyone involved m the neurosclences, both basic and clinical.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, 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 1s written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the widest range of s- drsciplines. Collecting these general techniques together in a single volume stnkes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of ammes and theirmetabolites and of amino acrds, respectively. These collections will clearly be of value to all neuroscientists working in or contemplating research in these fields.Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are provided for many types of ligand-receptor binding, including chapters on general prin- ples, drug discovery and development, and a most useful app- dix on computer programs for Scatchard, nonlinear, and compe- tive displacement analyses. Volume 5 provides procedures for the assessment of enzymes involved in biogenic amine synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physrologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, 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 neuroscientrsts and is written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the wrdest range of s- disciplines. Collecting these general techniques together in a single volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of amines and their metabolites and of amino acids, respectively. These collections will clearly be of value to all neuroscientists working in or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are provided for all types of ligand-receptor binding, including chapters on general principles, drug discovery and development, and a most useful-appendix on computer programs for Scatchard, nonlinear, and competitive d- placement analyses. Volume 5 provides procedures for the asse- ment of enzymes involved in biogenic amme synthesis and catabolrsm. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analyncal, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, 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 rt concerns those procedures used routmely across the wrdest range of s- disciplines. Collecting these general techniques together in a single volume strikes us not only as a service, but will no doubt prove of exceptronal utrlitarran value as well. Volumes 2 and 3 describe current procedures for the analyses of amines and their metabolites and of amino acids, respectively. These collections will clearly be of value to all neuroscrentists working m or contemplating research m these fields.Similar reasons exist for Volume 4 on receptor binding techniques, since experimental details are provided for many types of hgand-receptor binding, including chapters on general prin- ples, drug discovery and development, and a most useful app- dix on computer programs for Scatchard, nonlinear, and compe- tive displacement analyses. Volume 5 provides procedures for the assessment of enzymes involved in neurotransmitter synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologrsts, -anatomrsts, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved m the neuroscrences, both basic and clmical.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - 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 s- disciplines. Collecting these general techniques together in a single volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of amines and theirmetabolites and of amino acids, respectively. These collections will clearly be of value to all neurosclentists working m or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques, since experimental details are provided for all types of ligand-receptor binding, including chapters on general principles, drug discovery and development, and a most useful appendix on computer programs for Scatchard, nonlinear, and competitive d- placement analyses. Volume 5 provides procedures for the asse- ment of enzymes involved in biogenic amine synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomlsts, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

Recent years have seen remarkable advances in the devel- ment of techniques that have direct applications in neurological research. In consequence, the circulatory and metabolic status of the brain can be measured and correlated with changes m structure often noninvasively, m the same - and integrated function, perimental subIect This has stimulated an increased awareness of the complexity, under normal and pathological conditions, of the interdependence of these factors. Through the application of the methods described in this volume, however, these complexities can now be analyzed. The chapters m this volume present methodological - scriptions of some of the most powerful "physicochemical" methods for studymg the brain. Multidisciplmary teams are - quired to develop some of these methods, which are extremely expensive m terms of capital equipment costs and technological personnel support Thus, they will likely remain restricted to malor medical research centers Nevertheless, many recent concepts of brain responses to disease are a result of their application We have been fortunate m convincing active, leading sci- tists to contribute to this volume.The descrrptions of the basic prmciples of each method, and its applications and limitations, are derived primarily from their personal experiences. The first two chapters (Rowan, Auer) deal with methods for assessing brain hemodynamics. The two subsequent chapters (Greenberg; He- covitch) describe autoradiography and positron emission tomog- phy techniques, which provide quantitative measurements of brain metabolism as well as blood flow.

In this volume, we hope to cover the malor techniques that are presently being used to analyze the actions of drugs used in psyc- atry. The contributors of the chapters are active researchers who have considerable practical experience with the techniques they are describing, and the emphasis in the chapters is on three types of psychiatric drugs, namely antidepressants, antipsychotics, and anxiolytics. The first chapter, by Curry and Yu, discusses protein binding of psychotropic drugs, with special reference to equilibrium ana- sis as the method of assessment and to lipophilicity correlations. Since many of the drugs used in treating psychiatric disorders are bound extensively to protein, this aspect is of great importance with regard to their therapeutic actions and toxicity, Basic mat- matical models, techniques for the study of protein binding, molecular aspects of protein binding, binding in relation to lipophilicity, indirect approaches to measurement of the fraction of drug bound, the function of protein binding, and tissue binding are among the topics discussed. Chapter 2, by Norman and B- rows, deals with the principles of isotope derivative assays and their applications to antidepressants and antipsychotics; extensive protocols are provided. The third chapter, by Cooper, deals with analysis by gas-liquid chromatography. Sample collection and storage, extraction procedures, column selection, use of internal standards, types of detectors, techniques of application, and - plications to specific drugs are among the topics discussed.

Serendipitous discovery has played an important role m the identification of physiological mechanisms in central and peri- eral neurotransmission. In many cases, the subsequent followup of these discoveries has been substantially aided by previouslyiden- fied natural substances that interfered with these mechanisms. The site of neuromuscular blockade produced by curare extracts, for example, was shown in the 1850s by Claude Bernard, and Sir Henry H. Dale m 1906 discovered the reversal of the pressor effects of adrenal extracts by ergot alkaloids. These findings were the first clues that led eventually to the definition of mcotinic cholinergic receptors and the present classification of adrenergic receptors. In the 1950s many discoveries, including the identification of the catecholamines and 5-hydroxytryptamine in the central n- vous system, followed by the development of specific sensitive biochemical and histochemical methods and the introduction of modern psychotropic drugs led to our increased understanding of chemical transmission and the formulation of theories that attempt to explain the causes of some mental disorders.The finding that hydroxylated derivatives of noradrenaline and- hydroxytryptamine are readily taken up by catecholaminergic or serotonergic neurons, and that this is followed by poisoning and death of the cell, encouraged the use of these compounds for the mapping of central and peripheral catecholamine or- hydroxytryptamine systems, and the mvestigation of the re- vance of these pathways to receptor regulation or behavioral - sponses.

Lipids m the nervous system are major components of the m- branes. The presence of glycolipids in high concentrations is unique for the nervous system. Recent discoverres of the functronal and pathological importance of lipids such as diacylglycerols, polyphosphoinositides, prostaglandins, leukotrienes, docosahexaenoic acid, platelet activating factor, and gangliosides have markedly increased the number of publications on nervous system lipids. Many new methods have been developed. Thus, there IS a need for this volume that is dedicated to that methodo- gy. This volume places all of the methods for lipids into perspective with recommendations concerning the selection of a method for a specific purpose. Lloyd A. Horrocks vii Contents ...Preface to the Series vi Preface...*...xix List of Contributors Lipid Extraction Norman S. Radin 1 1. Introduction ...5 2. Properties of Solvents 2.1. Ethers ...6 8 2.2. Alcohols ...9 2.3. Halocarbons ...9 2.4. Hydrocarbons ...2.5. Miscellaneous Solvents ...10 ...10 3. Storage of Lipid Extracts 14 4. Homogenizers ...15 5. Separating the Extract from the Tissue Residue 15 5.1. Use of Filtration ...16 5.2. Centrifugation ...17 6.Removing Nonlipids from Lipid Extracts 6.1. Removal of Nonlipids by Evaporation and Reextraction ...17 ...17 6.2. Prewashing the Tissue Sample. 6.3. Removal of Nonlipids by Precipitating the Lipids with the Proteins ...18 ...19 6.4. Liquid/Liquid Partitioning ...19 6.5. Removal of Nonlipids by Dialysis 6.6. Separation from Nonlipids by Changing a 21 Partition Constant ...

The study of drug effects on behavior is truly a confluence of streams of neuroscience research. The material in this volume covers a broad range of methodology representative of practically the entire scope of psychopharmacology for conscious animals. The contributions in this book represent careful evaluations by experts who were invited to participate because of their excellent practical application of research design and methodology. In each case, a particularly useful and well-written contribution has been forthcoming, and should serve either as a valuable introduction to developing researchers or as a practical reference to those active in the field. The material is arranged in an intuitive hierarchy. This arrangement is based on what are ostensibly increasing levels of integrated activity of the nervous system and, therefore, increasing levels of behavioral integration. Nevertheless, as all of the areas covered are rather detailed in their levels of intrinsic complexity, this arrangement of chapters does not imply successively more intricate problems. The reader will find some overlap between chapters, but little interdependence, i.e., each may be read as a separate or as an integral part of the volume.

In order to understand how the human brain functions, one must first understand its structure and mechanics. This exciting collection of methods offers proven techniques for productive work in this fast-paced field. Contributors - all widely known for their mastery of the major methods described here - have been specially selected to share their knowledge in this volume.

Molecular Neurobiological Techniques explores the principles and techniques of molecular biology that have had a great impact on current investigations into the structure and function of the nervous system. It offers a solid basis for understanding how specific techniques are used to approach major neurobiological problems.

The mammalian central nervous system depends almost - clusively on glucose as its major energy source. In addition, g- cose participates in other cerebral metabolic functions including the biosynthesis of neurotransmitters, such as acetylcholine and the amino acids. This volume of Neuromethods assembles currently available methods for the study of cerebral glucose and energy metabolism in vitro and in mm. In the first chapter, Lust et al. describe the various methods available for the appropriate fixation of brain tissue necessary for the study of cerebral energy metabolism. Different fixation methods are compared, and some concerns raised by the USDHHS in their guidelines for the care and use of laboratory animals are addressed. Specific fixation methods pertinent to the various measurements are also covered in other chapters. In vitro p- parations have, despite certain limitations, been found to be useful in the study of brain metabolism, since the biochemical envir- ment is amenable to rapid, controlled manipulation.The chapter by Lai and Clark describes methods for the isolation and characterization of metabolically active preparations of synaptic and non-synaptic mitochondria from brain, and studies of - zymes involved in glucose metabolism and glucose-derived neurotransmitter synthesis in these preparations are summarized. The chapter by Whittingham discusses methods of preparations of hippocampal slices for use in the study of energy metabolism. Measurement of glucose and of glycolytic and dicarboxylic acid cycle intermediates in neural tissues are described in the chapter by Bachelard.

The latest information and methodologies for studying the activity of living nervous tissue, with special emphasis on electrophysiological techniques and their applications, ranging from ion channels to human behavior.

The wealth of material presented in Animal Models in Psychiatry has necessitated its expansion into a comprehensive, two-volume work. This second volume - the companion to Neuromethods 18 - continues the presentation of the latest methodologies for the use of animal models in psychiatric research. Contributors are renowned experts in their respective fields, and bring to light new insight into animal models for: performance deficits induced by stressors cholinergic supersensitivity and depression olfactory bulbectomy and depression assessment of circadian rhythms in affective disorders anxiety new anxiolytic drugs panicogenic and antipanic treatment effects drug studies in aggressive behavior mental retardation memory disorders. Boulton, Baker, and Martin-Iverson's two volumes on Animal Models in Psychiatry is the most current and comprehensive resource for everyone involved in the neurosciences, neuropsychiatry, pharmacology, and related disciplines.

The wealth of material presented in Animal Models in Psychiatry has necessitated its expansion into a comprehensive, two-volume work. This initial volume opens with an introductory chapter on criteria for assessing the validity of animal models in psychiatry, providing a framework in which to evaluate subsequent chapters in both volumes. Chapters are devoted to specific psychiatric disorders, along with various procedures that have been proposed as potential animal models for understanding their etiology and management. Contributors are renowned experts in their respective fields, and bring to light new insight into several animal models including: parallels to schizophrenia hippocampal lesions and schizophrenia stimulant psychoses hallucinations and continuous stimulants mania tardive dyskinesia activity anorexia attention deficit neuroleptic-induced oral dyskinesias. Everyone involved in psychiatric research, general neuroscience, and a host of other disciplines will find Boulton, Baker, and Martin-Iverson's timely new volume an indispensable resource for both office and laboratory.

This timely new volume in the highly acclaimed Neuromethods series focuses on major new advances in understanding the signaling mechanisms employed by neural tissues. Chapters, contributed by experts in the field, provide thorough, up-to-date coverage of a variety of topics and techniques, including: single-cell imaging technology phosphorylation of target proteins analysis of phosphoinositides and inositol phosphates inositol trisphosphate and intracellular calcium guanine nucleotide-binding proteins ion channel gating measurement of intracellular calcium with fluorescent calcium indicators protein kinase C synthetic analogs of intracellular messengers cyclic nucleotides caged intracellular messengers intracellular regulators and components of the exocytotic pathway intracellular messengers in vertebrate and invertebrate photoreceptors. Neuromethods 20 Intracellular Messengers is an essential resource for neurobiologiests at all levels of research.

The use of animal models continues to be the most reliable way of studying pathophysiologic mechanisms as well as treatments for neurological disorders. In two companion volumes, Animal Models of Neurological Disease highlights new information and techniques available in this rapidly growing area. This volume of Neuromethods focuses on commonly studied neurodegenerative diseases, highlighting the similarities and differences between the human condition and that obtained using animal models. Chapters are contributed by experts in the field and cover a variety of topics, including: the ethics of using animal models Alzheimer's disease Huntington's disease Parkinson s disease Lesch-Nyhan disease genetic and nongenetic demyelinating diseases cerebellar ataxias rodent and primate models surgical lesions neurotoxins. Boulton, Baker, and Butterworth's important new treatise on Animal Models of Neurological Disease is an outstanding guide for all researchers striving for a better understanding of neurological disease.

The use of animal models continues to be the most reliable way of studying pathophysiologic mechanisms as well as treatments for neurological disorders. In two companion volumes, Animal Models of Neurological Disease highlights new information and techniques available in this rapidly growing area. This volume of Neuromethods - the second of a two-volume set on Animal Models of Neurological Disease - presents forefront techniques for studying a variety of seizure disorders and metabolic diseases affecting the CNS. Contributions have been gleaned from a panel of experts in several areas, including: human stroke focal and global ischemia brain hypoxia in vivo and in vitro Wernicke-Korsakoff syndrome chemically induced and genetic-based seizures the kindling model of epilepsy hepatic encephalopathies vitamin deficiency syndromes hereditary hyperammonemias. Boulton, Baker, and Butterworth's important new set on Animal Models of Neurological Disease is an outstanding guide for all researchers striving to better understand neurological disease.

Practical Cell Culture Techniques focuses on the advantages of new tissue culture techniques that enable close and detailed study of many nerve cell types. Neuroscience is one of the key areas whose rapid development has been driven by the proliferation of cell culture methodology; some areas of neuroscience, such as glial cell biology and function, now depend almost entirely on cell culture-based research projects. This latest volume of Humana's renowned Neuromethods series provides practical treatment of the new, powerful methodologies and results including: cell culture techniques specifically designed for the novice the environmental factors that play a role in tissue culturing cell markers nutrition requirements of the different cell types three-dimensional organotypic slice explant cultures neuronal cultures cultures of astrocytes, oligodendrocytes, cerebral capillary endothelium, and other cell types. Boulton, Baker, and Walz's new volume on Practical Cell Culture Techniques is sure to be a major resource for everyone - whether expert or novice - involved in studying the central nervous system on the molecular level.

A comprehensive guide to the use of animal models for understanding the many facets of drug and alcohol abuse - problems that are now worldwide health concerns. Expert investigators have contributed state-of-the-art chapters on the use of animal models for research on substance abuse.

Leading researchers present those techniques that have been crucial to propelling the neurotrophic factors field to its current state of high visibility. The techniques range from molecular biological methods used for cloning and production, to cell culture methods for assessing biological activities, to animal models of nervous system injury (necessary for the development of therapeutic agents from neurotrophic factors).

Patch-Clamp Applications and Protocols presents the broadest coverage of techniques for measuring and recording directly activity within a cell. The first part of the book offers modern developments associated with the technology of path-clamp electrodes, of cell-free ion-channel recording, and of the whole-cell patch clamp technique. Chapters on such recent offspring of the technique as the concentration clamp technique, the pressure clap method, and the perfusion of patch clamp electrodes are contributed by authors intimately involved with their development. Also covered is the loose patch-clamp technique, single-channel recording, the perforated patch-clamp technique, molecular biological aspects of the technique, and the use of the technique in brain slices. Every scientist working with path-clamp methods will find this protocols manual invaluable to their continued use and development of this powerful technique.

Efforts to measure neurotransmitters and related s- cies in living brain tissue by faradic electrochemistry began in earnest in the early 1970s. During the ensuing years, s- eral monographs and various symposia on the subject have appeared. The present volume of Neuromethods, Voltammetric Methods in Brain Systems, can be considered different from previous offerings in several respects. The methodology of in vivo electrochemistry has now reached a stable level of maturity. It is no longer necessary to convince interested re- ers that the techniques can measure, with specified degrees of reliability, certain neurotransmitters and metabolites in the extracellular fluid space. Moreover, the basic approaches of electroanalytical measurements are more widely appre- ated by neuroscientists and one need not explain in tutorial fashion the details of oxidative electrochemistry. Nevert- less, in keeping with the nature of the Neurumethods series, this is a "how to" volume and readers will find ample detail on the preparation of electrodes, practical experimental details, and the interpretation of results. But the emphasis is strongly on the application of voltammetric methods.The first two chapters deal with fundamentals inherent in employing in vivo electrochemical measures. The first - ticle offers an authoritative account of our understanding of the surface states of carbon and carbon fiber electrodes. It provides a rationale for the various pretreatments that have allowed the "tuning" of electrode properties to obtain sen- tivity and fast response times.

Practical Cell Culture Techniques focuses on the advantages of new tissue culture techniques that enable close and detailed study of many nerve cell types. Neuroscience is one of the key areas whose rapid development has been driven by the proliferation of cell culture methodology; some areas of neuroscience, such as glial cell biology and function, now depend almost entirely on cell culture-based research projects. This latest volume of Humana's renowned Neuromethods series provides practical treatment of the new, powerful methodologies and results including: cell culture techniques specifically designed for the novice the environmental factors that play a role in tissue culturing cell markers nutrition requirements of the different cell types three-dimensional organotypic slice explant cultures neuronal cultures cultures of astrocytes, oligodendrocytes, cerebral capillary endothelium, and other cell types. Boulton, Baker, and Walz's new volume on Practical Cell Culture Techniques is sure to be a major resource for everyone - whether expert or novice - involved in studying the central nervous system on the molecular level.

The G proteins are a family of structurally homologous, plasma membrane-associated guanine-nucleotide-binding proteins. These proteins play an integral role in the tra- duction of extracellular signals through second messenger systems. As such, G proteins affect a wide variety of intra cellular biochemical reactions by regulating the concent- tion of second messengers in cells. G proteins are heterotrimeric, consisting of a, p, and y polypeptide chains, with G protein specificity largely det- mined by the a-subunit, Molecular cloning of G protein s- units has revealed 23 distinct a-subunits, encoded by 17 different genes. Based on functional measures, G proteins are generally classified into three major categories: the G, family, which is stimulatory for adenylyl cyclase; the G, f- ily, which is inhibitory for adenylyl cyclase; and the G, f- ily, which stimulates phospholipases (Birnbaumer and Birnbaumer, 1995). Alternatively, on the basis of sequence homology, G proteins can be subdivided into four cate- ries: G,, G,, G,, and G12.