Protocols and Good Operating Practices in the Study of Cannabinoid Receptors

With the approach of the 30th year since the pioneering discovery of a cannabinoid receptor in rat brain (Devane et al., 1988), the field of cannabinoid pharmacology and physiology has impacted human physiology at multiple levels. The development of highly specific and potent orthosteric ligands, as well as the blossoming field of allosteric ligand development, has placed the endocannabinoid system in the forefront as a modulator of a multitude of physiologic processes. Reproducibility among laboratories is especially important due to the development of novel tools to investigate the role(s) of the endocannabinoid system in human physiology, and to clarify the roles for medicinal marijuana. Any definitive role in normal, or diseased states, must be satisfied through the demonstration of a specific receptor-mediated event. This chapter provides working protocols for the study of cannabinoid receptor-ligand binding, as well as immediate and downstream G protein-dependent signaling assays to assess receptor function.