This
volume encompasses all major methodologies to interrogate endocannabinoid
systems (ECS) and endocannabinoids (eCBs) signaling. With increasing interest
towards the manifold activities of eCBs, this book discusses the chemical,
biochemical, and molecular biological assays, and activity of distinct elements
of the ECS. These include membrane, nuclear receptors, biosynthetic and
hydrolytic enzymes, and membrane transporters and oxidative enzymes. 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, readily reproducible laboratory
protocols, and tips on troubleshooting and avoiding known pitfalls.
Timely and cutting edge, Endocannabinoid Signaling: Methods and Protocols
is a valuable resource and will help chemists, drug designers, biochemists,
molecular biologists, cell biologists, pharmacologists, and (electro)
physiologists navigate the mare magnum of endocannabinoid research.
1. Need for Methods to Investigate
Endocannabinoid Signaling
Mauro Maccarrone
2. Extraction and Simultaneous Quantification
of Endocannabinoids and Endocannabinoid-Like Lipids
in Biological Tissues
Laura Bindila and Beat Lutz
3. Determination of 2-Arachidonoylglycerol by µSPE-LC-MS/MS
Natalia Battista and Manuel
Sergi
4. Analysis of Omega-3 Fatty Acid
Derived N-acylethanolamines in Biological
Matrices
Renger F. Witkamp and Michiel G.J.
Balvers
5. Assay of CB1 Receptor Binding
Valeria Catani and Valeria Gasperi
6. The Displacement Binding Assay Using Human
Cannabinoid CB2 Receptor-Transfected Cells
Maria Grazia Cascio, Pietro Marini, and
Roger G. Pertwee
7. Assay of TRPV1 Receptor Signaling<
Aniello Schiano Moriello and Luciano
De Petrocellis
8. A Functional Assay for GPR55: Envision Protocols
Sharon Anavi-Goffer and Ruth A. Ross
9. The Cyclic AMP Assay Using Human Cannabinoid
CB2 Receptor-Transfected Cells
Pietri Marini, Maria Grazia Cascio, and
Roger G. Pertwee
10. Assay of GTPγS Binding in Autoradiography
Marina Gabaglio,
Pamela Prini, Erica Zamberletti, Tiziana Rubino, and Daniela Parolaro
11. Protocol to Study β-Arrestin Recruitment by CB1 and CB2
Cannabinoid Receptors
Marjolein Soethoudt, Noortje van
Gils, Mario van der Stelt, and Laura H. Heitman
12. Assay of NAT Activity
Toru Uyama and Matsuo Ueda
13. Assay of NAPE-PLD Activity
Filomena Fezza, Nicolina Mastrangelo,
and Mauro Maccarrone
14. Assay of FAAH Activity
Monica Bari, Monica Feole, and Mauro
Maccarrone
15. Assay of NAAA Activity
Kazuhito Tsuboi and Natsuo Ueda
16. Assay of DAGLα/β Activity
Tiziana Bisogno
17. Assay of Monoacylglycerol Lipase Activity
Kwang-Mook Jung and Daniele Piomelli
18. A Sensitive and Versatile Fluorescent Activity Assay for ABHD6
Juha R. Savinainen, Dina
Navia-Paldanius, and Jarmo T. Laitinen
19. A Sensitive and Versatile Fluorescent Activity Assay for ABHD12
Juha R. Savinainen, Dina
Navia-Paldanius, and Jarmo T. Laitinen
20. Assay of Endocannabinoid Uptake
Mark Ray, Simon Nicolussi, Andreas
Chicca, and Jürg Gertsch
21. Assay of Endocannabinoid Oxidation by Cyclooxygenase-2
Shalley N. Kudalkar, Philip J.
Kingsley, and Lawrence J. Marnett
22. Oxygenation of Anandamide by Lipoxygenases
Guus van Zadelhoff and Mario van der
Stelt
23. Assay of Endocannabinoid Oxidation by Cytochrome P450
Natasha T. Snider, Vyvyca J.
Walker, and Paul F. Hollenberg
24. Assessing Gene Expression of the Endocannabinoid System
Mariangela Pucci and Claudio
D’Addario
25. Western Blotting of the Endocannabinoid System
Jim Wager-Miller and Ken Kackie
26. Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Trafficking
in Cultured Cells
Jim Wager-Miller and Ken Mackie
27. Measuring ECS Interaction with Biomembranes
Clotilde B. Angelucci, Annalaura
Sabatucci, and Enrico Dainese
28. Visualization of Endocannabinoids in the Cell
Sergio Oddi, Antonio Totaro, and
Mauro Maccarrone
This
volume encompasses all major methodologies to interrogate endocannabinoid
systems (ECS) and endocannabinoids (eCBs) signaling. With increasing interest
towards the manifold activities of eCBs, this book discusses the chemical,
biochemical, and molecular biological assays, and activity of distinct elements
of the ECS. These include membrane, nuclear receptors, biosynthetic and
hydrolytic enzymes, and membrane transporters and oxidative enzymes. 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, readily reproducible laboratory
protocols, and tips on troubleshooting and avoiding known pitfalls.
Timely and cutting edge, Endocannabinoid Signaling: Methods and Protocols
is a valuable resource and will help chemists, drug designers, biochemists,
molecular biologists, cell biologists, pharmacologists, and (electro)
physiologists navigate the mare magnum of endocannabinoid research.
Includes cutting-edge methods and protocols
Provides step-by-step detail essential for reproducible results
Contains key notes and implelementation advice from the experts