The endogenous cannabinoid anandamide is an agonist of the cannabinoid receptors CB1 and CB2, as well as transient receptor potential vanilloid type 1 and type 4 ion channels. In recent years, there has been a great deal of interest in the cellular processes regulating the signaling of endocannabinoids such as anandamide. This is due largely to evidence that augmentation of cannabinergic tone might be therapeutically beneficial in the treatment of multiple disease states such as chronic pain, anxiety, multiple sclerosis, and neuropsychiatric disorders. Of particular interest are the cellular processes that regulate the cellular accumulation and metabolism of anandamide. Characterization of the process by which anandamide is internalized and metabolized by the cell may identify drug targets useful in the positive modulation of cannabinergic tone. Recently, we reported that detergent-resistant membrane microdomains known as lipid rafts play a role in the cellular accumulation of anandamide by mediating an endocytic process responsible for anandamide internalization. The enzyme primarily responsible for anandamide metabolism, fatty acid amide hydrolase, is excluded from lipid rafts. However, the metabolites of anandamide accumulate in these detergent-resistant membrane microdomains. There is some preliminary evidence that makes it reasonable to propose that anandamide metabolites enriched in lipid rafts may act as precursors to anandamide synthesis. Overall, experimental evidence is mounting that detergent-resistant membrane microdomains such as lipid rafts may play a role in the cellular regulation of anandamide inactivation and production.