Main description:

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 (TM) 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.

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, Cherine 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 Graslund, 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

17. Antibody Microprobes for Detecting Neuropeptide Release

Carole A. Williams and Arthur W. Duggan

18. Neuropeptidases

Manuel Ramirez, Isabel Prieto, Inmaculada Banegas, Ana B. Segarra, and Francisco Alba

19. Neuropeptide Autoantibodies Assay

Serguei 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

22. Measurement of Phosphorothioate Oligodeoxynucleotide Antisense Transport Across the Blood-Brain Barrier

William A. Banks

23. Liposome-Encapsulated Neuropeptides for Site-Specific Microinjection

Frederic Frezard, 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