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.

Table of contents 1. What Are Neuropeptides?
J. Peter H. Burbach
2. Neuropeptide Localization in Non Mammalian Vertebrates
Paolo de Girolamo and Carla Lucini
3. Combined Light and Electron Microscopic Visualization of Neuropeptides and Their Receptors in Central Neurons
Chiara Salio, Laura Lossi, and Adalberto Merighi
4. Neuropeptide RNA Localization in Tissue Sections
Marc Landry, Chérine Abdel Salam, and Marie Moftah
5. Intron-Specific Neuropeptide Probes
Harold Gainer, Todd A. Ponzio, Chunmei Yue, and Makoto Kawasaki
6. Direct In Situ RT-PCR
Laura Lossi, Graziana Gambino, Chiara Salio, and Adalberto Merighi
7. Laser Capture Microdissection and Quantitative-PCR Analysis
Sarah J. Paulsen and Leif K. Larsen
8. Visualization of Peptide Secretory Vesicles in Living Nerve Cells
Park J. Joshua and Loh Y. Peng
9. Fluorescence Imaging with Single-Molecule Sensitivity and Fluorescence Correlation Spectroscopy (FCS) of Cell-Penetrating Neuropeptides
Vladana Vukojevic, Astrid Gräslund, and Georgy Bakalkin
10. Analysis of Neuro-Immune Interactions by an In Vitro Co-Culture Approach
Tadahide Furuno and Mamoru Nakanishi
11. Electrophysiology
Zhi-Qing David Xu
12. Localization of Neuropeptides by Radioimmunoassay
Fred Nyberg and Mathias Hallberg
13. Reversed-Phase HPLC and Hyphenated Analytical Strategies for Peptidomics
Anne-Marie Hesse, Sega Ndiaye, and Joelle Vinh
14. Neuropeptidomics: Mass Spectrometry-Based Qualitative and Quantitative Analysis
Ping Yin, Xiaowen Hou, Elena V. Romanova, and Jonathan V. Sweedler
15. Suppression Subtractive Hybridization
Mohamed T. Ghorbel and David Murphy
16. Neuropeptide Microdialysis in Free-Moving Animals
Tetsuya Kushikata and Kazuyoshi Hirota
7. Antibody Microprobes for Detecting Neuropeptide Release
Carole A. Williams and Arthur W. Duggan
18. Neuropeptidases
Manuel Ramírez, Isabel Prieto, Inmaculada Banegas, Ana B. Segarra, and Francisco Alba
19. Neuropeptide Autoantibodies Assay
Sergueï O. Fetissov
20. Intranasal Delivery of Neuropeptides
Michael C. Veronesi, Daniel J. Kubek, and Michael J. Kubek
21. Prodrug Design for Brain Delivery of Small- and Medium-Sized Neuropeptides
Katalin Prokai-Tatrai and Laszlo Prokai
2. Measurement of Phosphorothioate Oligodeoxynucleotide Antisense Transport Across the Blood-Brain Barrier
William A. Banks
23. Liposome-Encapsulated Neuropeptides for Site-Specific Microinjection
Frédéric Frézard, Robson A.S. dos Santos, and Marco A.P. Fontes
24. Recombinant Adeno-Associated Viral Vectors
Marijke W.A. de Backer, Keith M. Garner, Mieneke C.M. Luijendijk, and Roger A.H. Adan
25. De-Orphanizing G Protein-Coupled Receptors by a Calcium Mobilization Assay
Isabel Beets, Marleen Lindemans, Tom Janssen, and Peter Verleyen