Main description:

This second edition provides new and updated chapters detailing all major elements of the ECB system. Chapters guide readers through identification of drug targets, electrophysiology, computational chemistry, and machine learning. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Endocannabinoid Signaling: Methods and Protocols, Second Edition is a valuable resource for all researchers interested in learning more about this important and developing field.

Contents:

1. Need for Methods to Investigate Endocannabinoid Signaling

Mauro Maccarrone

2. Extraction and Simultaneous Quantification of Endocannabinoids and Endocannabinoid-like Lipids in Biological Tissues

Claudia Schwitter, Beat Lutz, and Laura Bindila

3. Measuring the Content of Endocannabinoid-like Compounds in Biological Fluids: A Critical Overview of Sample Preparation Methodologies

Heather B Bradshaw and Clare T Johnson

4. LC-MS/MS Analysis of AEA and 2-AG

Natalia Battista, Federico Fanti, and Manuel Sergi

5. Analysis of Omega-3 Fatty Acid Derived N-Acylethanolamines in Biological Matrices

Renger F. Witkamp, Ian de Bus, Bauke Albada, and Michiel G.J. Balvers

6. Endocannabinoid-Binding Receptors as Drug TargetsMaria Gomez-Canas, Carmen Rodriguez-Cueto, Valentina Satta, Ines Hernandez-Fisac, Elisa Navarro, and Javier Fernandez-Ruiz

7. Assay of CB1 Receptor Binding

Valeria Gasperi, Isabella Savini and Maria Valeria Catani

8. The Displacement Binding Assay Using Human Cannabinoid CB2 Receptor-Transfected CellsMaria Grazia Cascio, Pietro Marini and Roger G. Pertwee

9. Fluorescence-based Assay for TRPV1 Channels

Aniello Schiano Moriello, Luciano De Petrocellis, Rosa Maria Vitale

10. DNA-Protein-Interaction (DPI)-ELISA Assay for PPAR- Receptor Binding

Maria Valeria Catani, Valentina Tullio, Mauro Maccarrone, and Valeria Gasperi

11. Scintillation Proximity Assay (SPA)-Based Radioligand Binding for PPAR , PPAR and PPAR ReceptorsUwe Grether, Joerg Benz, Thomas Hartung, and Doris Roth

12. A Homogeneous Time-Resolved Fluorescence Resonance Energy Transfer (TR-FRET) Cofactor Recruitment Assay for PPAR and PPAR Doris Roth, Joerg Benz, Uwe Grether, and Michel Dietz

13. The Cyclic AMP Assay Using Human Cannabinoid CB2 Receptor-Transfected Cells

Pietro Marini, Maria Grazia Cascio, and Roger G. Pertwee

14. Assay of GTP S Binding in Autoradiography

Marina Gabaglio, Pamela Prini, Erica Zamberletti, Tiziana Rubino, and Daniela Parolaro

15. Cellular Assay to Study -Arrestin Recruitment by the Cannabinoid Receptors 1 and 2

Jara Bouma, Marjolein Soethoudt, Noortje van Gils1, Lizi Xia1, Mario van der Stelt2, 3 and Laura H. Heitman

16. Endocannabinoid Metabolism and Transport as Drug Targets

Francesca Ciaramellano, Federico Fanti, Lucia Scipioni, Mauro Maccarrone, and Sergio Oddi

17. Assay of NAT Activity

Toru Uyama and Natsuo Ueda

18. Radiometric Assay of NAPE-PLD activity

Filomena Fezza, Nicolina Mastrangelo, and Mauro Maccarrone

19. A Fluorescence-based NAPE-PLD Activity Assay

Elliot D. Mock, Wouter P. F. Driever, and Mario van der Stelt

20. Radiometric Assay of FAAH Activity

Monica Bari, Monica Feole, Marina Fava, and Mauro Maccarrone

21. Fluorimetric Assay of FAAH activityClotilde B. Angelucci, Roberto Giacominelli-Stuffler,and Mauro Maccarrone

22. Assay of NAAA Activity

Kazuhito Tsuboi and Natsuo Ueda

23. Assay of DAGL / Activity

Tiziana Bisogno

24. Assay of Monoacylglycerol Lipase Activity

Kwang-Mook Jung and Daniele Piomelli

25. Radiometric Assay of ABHD2 Activity

Domenico Fazio, Emanuele Criscuolo, and Mauro Maccarrone

26. Oxygenation of Anandamide by Lipoxygenases

Guus van Zadelhoff and Mario van der Stelt

27. Assay of Endocannabinoid Oxidation by Cytochrome P450

Natasha T. Snider and Paul F. Hollenberg

28. Assays of Endocannabinoid Uptake

Ines Reynoso-Moreno, Mark Rau, Andrea Chicca, Simon Nicolussi, and Jurg Gertsch

29. Assessing Gene Expression of the Endocannabinoid System Components by Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction

Mariangela Pucci and Claudio D'Addario

30. Bioinformatics of the Endocannabinoid System: Study of DNA methylation at Rat Cnr1 gene promoterAnnalaura Sabatucci and Claudio D'Addario

31. DNA Methylation Analysis of Cnr1 Gene Promoter

Claudio D'Addario and Mariangela Pucci

32. Western Blotting of the Endocannabinoid System

Jim Wager-Miller and Ken Mackie

33. Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Trafficking in Cultured Cells

Jim Wager-Miller and Ken Mackie

34. Assessing CB1 Expression in the Brain by Immunohistochemical Methods: Light, Confocal and Electron Microscopy

Roberta Imperatore and Luigia Cristino

35. Measuring Endocannabinoid System Interaction with Biomembranes

Clotilde B. Angelucci, Annalaura Sabatucci, Ana Lia Bernardo, Alexandrine Kurtz, Sergio Oddi, and Enrico Dainese

36. STORM Super-Resolution Imaging of CB1 Receptors in Tissue Preparations

Miklos Zoeldi and Istvan Katona

37. Visualization of Endocannabinoids in the Cell

Sergio Oddi, Francesca Ciaramellano, Lucia Scipioni, Enrico Dainese, and Mauro Maccarrone

38. Electrophysiology of Endocannabinoid Signalling

Alessandra Musella and Diego Centonze

39. Machine Learning and Computational Chemistry for the Endocannabinoid System

Kenneth Atz, Wolfgang Guba, Uwe Grether, and Gisbert Schneider

40. A Virtual Reverse Screening Approach to Target Type 2 Cannabinoid Receptor

Fulvio Ciriaco, Nicola Gambacorta, Francesco Leonetti, Cosimo Damiano Altomare,and Orazio Nicolotti