Rapid advances in our understanding of basic cell biological processes and of the molecular mechanisms of cell function and dysfunction have led to an increasing interest in utilizing these approaches in neurobiological research. Efforts in the most rapidly advancing areas are multidisciplinary and the o- come of contributions from many investigators employing a variety of te- niques to address a specific problem. Although a strong basis in fundamental neurobiological concepts is essential for each researcher, the ability to apply new techniques and approaches to the examinination of both cellular and molecular processes requires knowledge of a wide variety of methodologies. The objective of Neurodegeneration Methods and Protocols is to develop an understanding, appreciation, and technical ability in various cellular and molecular techniques for studying many aspects of nervous system cell bi- ogy. The protocols in this volume span a multidisciplinary range of cellular and molecular approaches, and should allow investigators to address research questions directed toward understanding nervous system function, injury, degeneration, and the repair/regenerative process.

Klappentext

An outstanding collection of detailed, state-of-the-art techniques for studying many aspects of nervous system cell biology. The methods span a multidisciplinary range of cellular and molecular approaches to both normal and injured nervous system function, and particularly to neurodegenerative processes. Included are basic RT-PCR techniques, cell culture systems, second-messenger signaling methods, and patch-clamp techniques. These powerful techniques will illuminate nervous system function, injury, degeneration, and the repair/regenerative process.

Inhalt
to Polymerase Chain Reaction.- Methodological Issues in Polymerase Chain Reaction for RNA.- Subtractive Hybridization of cDNA Libraries.- The Use of Differential Display RT-PCR for Identifying Altered Gene Expression in Ischemic Brain Injury.- Analysis of Gene Expression by Multiprobe RNase Protection Assay.- Molecular Probes for PNS Neurotoxicity, Degeneration, and Regeneration.- Mobility-Shift DNA-Binding Assay Using Gel Electrophoresis.- Transfection of Mammalian Cells In Vitro: Use in Analysis of Neuronal Damage.- Detection of Apoptotic Cells in the Nervous System.- Translocation Assays of Protein Kinase C Activation.- Astrocyte and Neuron Coculturing Method.- Viability Assays for Cells In Vitro: The Ethidium/Calcein Assay and the Immunofluorescence Combination Assay.- Measurement of the Nitric Oxide Synthase Activity Using the Citrulline Assay.- Measurement of Intraneuronal Free Calcium Using the Fluorescent Probe Technique.- Measurement of Calcium Buffering by Intracellular Organelles in Brain.- Receptor-Mediated Release of Inositolphosphates in Brain Slices.- The Use of Enzyme-Linked Immunosorbent Assays to Quantitate Proteins in Biological Functions.- Assays for the Analysis of Synaptic Proteins in Neurodegenerative Disorders.- Zymographic Method for the Measurement of Gelatinase Activity in Brain Tissue.- The Measurement of Monoamine Neurotransmitters in Microdialysis Perfusates Using HPLC-ECD.- Biochemical Measurement of Cholinesterase Activity.- Functional Study of Glutamate Receptor Channels in Brain Slices.- Selective Silver Staining for Neural Degeneration.- Immunohistological Markers of Cerebrovascular Integrity.- Electron Probe X-Ray Microanalysis: A Quantitative Electron Microscopy Technique for Measurement of Elements and Water in Nervous Tissue Cells.