Neuropeptides

This book presents a readily reproducible collection of established and emerging techniques for neuropeptide research. Coverage includes immunocytochemical localization, biochemical characterization, development of genetic probes and functional analysis.

Since the early days, the field of neuropeptide biology has dramatically widened, and today the ultimate frontiers in neuropeptide research lie in the development of pharmacologically active compounds that are capable of crossing the blood-brain barrier to exert their biological role(s) in vivo and in the construction of genetic vectors to be employed in gene therapy. Neuropeptides: Methods and Protocols presents a readily reproducible collection of established and emerging techniques for neuropeptide research as contributed by expert researchers in the field. The detailed methods presented cover subjects such as immunocytochemical localization, biochemical characterization, functional analysis, development and production of genetic probes, and the design of neuropeptide derivatives for cellular neurobiology as well as the potential therapeutic applications. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

Authoritative and easy to follow, Neuropeptides: Methods and Protocols is intended for a large audience of scientists, including histologists, biochemists, cellular and molecular biologists, and electrophysiologists that are currently active in the field or are willing to enter this exciting and still expanding area of neurobiology.

Features contributions from scientists with direct experience in one or more of the fields of neuropeptide research Include detailed protocols relevant to a wide range of scientists Provides key tips and expert implementation advice to ensure successful results

Inhalt

What Are Neuropeptides?.- Neuropeptide Localization in Non Mammalian Vertebrates.- Combined Light and Electron Microscopic Visualization of Neuropeptides and Their Receptors in Central Neurons.- Neuropeptide RNA Localization in Tissue Sections.- Intron-Specific Neuropeptide Probes.- Direct In Situ RT-PCR.- Laser Capture Microdissection and Quantitative-PCR Analysis.- Visualization of Peptide Secretory Vesicles in Living Nerve Cells.- Fluorescence Imaging with Single-Molecule Sensitivity and Fluorescence Correlation Spectroscopy (FCS) of Cell-Penetrating Neuropeptides.- Analysis of Neuro-Immune Interactions by an In Vitro Co-Culture Approach.- Electrophysiology.- Localization of Neuropeptides by Radioimmunoassay.- Reversed-Phase HPLC and Hyphenated Analytical Strategies for Peptidomics.- Neuropeptidomics: Mass Spectrometry-Based Qualitative and Quantitative Analysis.- Suppression Subtractive Hybridization.- Neuropeptide Microdialysis in Free-Moving Animals.- Antibody Microprobes for Detecting Neuropeptide Release.- Neuropeptidases.- Neuropeptide Autoantibodies Assay.- Intranasal Delivery of Neuropeptides.- Prodrug Design for Brain Delivery of Small- and Medium-Sized Neuropeptides.- Measurement of Phosphorothioate Oligodeoxynucleotide Antisense Transport Across the Blood-Brain Barrier.- Liposome-Encapsulated Neuropeptides for Site-Specific Microinjection.- Recombinant Adeno-Associated Viral Vectors.- De-Orphanizing G Protein-Coupled Receptors by a Calcium Mobilization Assay.