Neurotransmitter Methods

Neurotransmitter Methods is intended as a bench-side companion for researchers who seek to identify, locaHze, or measure neurotransmitters and/or to identify sites of neurotransmitter action. Each method is detailed in a user-friendly "recipe" format and the protocols are accompanied by extensive notes to high light and explain crucial steps. Approaches utilizing an incredibly diverse array of modem techniques are presented: methods including HPLC, histochemistry, immunocytochemistry, in situ hybridization, mass spectrometry, microdialysis, and electrochemistry all make at least one appearance. In addition, protocols for associated methodologies, including the production of brain slices, dissociated neurons, synaptosomes/synaptoneurosomes, and neuronal plasma membranes are presented. Methods applicable to most of the recognized chemical types of neu rotransmitter are to be found and, although you may find absent any mention of your favorite neurotransmitter, many of the protocols are sufficiently general to be adapted to alternative uses. So, how does this book contribute usefiilly to the horde of methods volumes stampeding across our bookshelves? 1 hope that the strength oiNeurotransmitter Methods lies in the variety of its content. The book provides in a single volume an array of techniques that could take a researcher from selection and preparation of a tissue source through to identification and measurement of neurotransmitter content and even onto characterization of neurotransmitter sites of action. Furthermore, the diversity of approaches presented reflects the need for today's bench neuroscientist to be a bit of a "jack (or a "jill"!) of all trades.

Klappentext

This outstanding and comprehensive collection of user-friendly protocols is essential for neuroscientists who wish to localize, identify, purify, and/or quantify neurotransmitters. The easily reproducible methods presented cover a broad spectrum of approaches involving cell and organ culture, histochemistry, immunochemistry, in situ hybridization, cell fractionation, liquid chromatography, mass spectrometry, in vivo microdialysis, and voltammetry. These approaches are optimized for use by neuroscientists, are often complementary, and can be employed in tandem with powerful effect.

Inhalt
Preparation of Brain Slices.- Preparation of Organotypic Hippocampal Slice Cultures Using the Membrane Filter Method.- The Preparation and Use of Brain Slices and Dissociated Neurons for Patch-Clamp Studies of Neurotransmitter Action.- Synaptosomes.- Synaptoneurosomes.- Preparation of Synaptosomal Plasma Membranes by Subcellular Fractionation.- Detection of Neuropeptides by Immunocytochemistry.- Immunocytochemical Detection of Amino Acid Neurotransmitters in Paraformaldehyde-Fixed Tissues.- cGMP-Immunocytochemistry.- Microwave Antigen Retrieval in Formaldehyde-Fixed Human Brain Tissue.- Localization of Nitric Oxide Synthase Using NADPH Diaphorase Histochemistry.- In Situ Hybridization to Determine the Expression of Peptide Neurotransmitters.- Quantitative In Situ Hybridization Histochemistry.- Microbore Liquid Chromatography Analysis of Monoamine Transmitters.- Microbore Liquid Chromatography Analysis of Amino Acid Transmitters.- HPLC Methods to Isolate Peptide Neurotransmitters.- Neuropeptide Expression Patterns as Determined by Matrix-Assisted Laser Desorption Ionization Mass Spectrometry.- GC/MS Determination of Biogenic Amines in insect Neurons.- Monitoring Amino Acid Neurotransmitter Release in the Brain by In Vivo Microdialysis.- In Vivo Detection of Neurotransmitters with Fast Cyclic Voltammetry.