Endocannabinoid Signaling

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

Inhalt

Need for Methods to Investigate Endocannabinoid Signaling.- Extraction and Simultaneous Quantification of Endocannabinoids and Endocannabinoid-Like Lipids in Biological Tissues.- Determination of 2-Arachidonoylglycerol by µSPE-LC-MS/MS.- Analysis of Omega-3 Fatty Acid Derived N -acylethanolamines in Biological Matrices.- Assay of CB 1 Receptor Binding.- The Displacement Binding Assay Using Human Cannabinoid CB 2 Receptor-Transfected Cells.- Assay of TRPV1 Receptor Signaling.- A Functional Assay for GPR55: Envision Protocols.- The Cyclic AMP Assay Using Human Cannabinoid CB 2 Receptor-Transfected Cells.- Assay of GTPS Binding in Autoradiography.- Protocol to Study -Arrestin Recruitment by CB 1 and CB 2 Cannabinoid Receptors.- Assay of NAT Activity.- Assay of NAPE-PLD Activity.- Assay of FAAH Activity.- Assay of NAAA Activity.- Assay of DAGL/ Activity.- Assay of Monoacylglycerol Lipase Activity.- A Sensitive and Versatile Fluorescent Activity Assay for ABHD6.- A Sensitive and Versatile Fluorescent Activity Assay for ABHD12.- Assay of Endocannabinoid Uptake.- Assay of Endocannabinoid Oxidation by Cyclooxygenase-2.- Oxygenation of Anandamide by Lipoxygenases.- Assay of Endocannabinoid Oxidation by Cytochrome P 450.- Assessing Gene Expression of the Endocannabinoid System.- Western Blotting of the Endocannabinoid System.- Quantitation of Plasma Membrane (G Protein-Coupled) Receptor Trafficking in Cultured Cells.- Measuring ECS Interaction with Biomembranes.- Visualization of Endocannabinoids in the Cell.