title: Characterization of biotin-anandamide, a novel tool for the visualization of anandamide accumulation.
abstract: Anandamide (N-arachidonoylethanolamide; AEA) acts as an endogenous agonist of both cannabinoid and vanilloid receptors. During the last two decades, its metabolic pathways and biological activity have been investigated extensively and relatively well characterized. In contrast, at present, the effective nature and mechanism of AEA transport remain controversial and still unsolved issues. Here, we report the characterization of a biotinylated analog of AEA (b-AEA) that has the same lipophilicity of the parent compound. In addition, by means of biochemical assays and fluorescence microscopy, we show that b-AEA is accumulated inside the cells in a way superimposable on that of AEA. Conversely, b-AEA does not interact or interfere with the other components of the endocannabinoid system, such as type-1 and type-2 cannabinoid receptors, vanilloid receptor, AEA synthetase (N-acylphosphatidylethanolamine-hydrolyzing phospholipase D), or AEA hydrolase (fatty acid amide hydrolase). Together, our data suggest that b-AEA could be a very useful probe for visualizing the accumulation and intracellular distribution of this endocannabinoid.
authors.affiliation: European Center for Brain Research/Istituto di Ricovero e Cura a Carattere Scientifico S. Lucia Foundation, Rome, Italy.
authors.forename: Filomena
authors.initials: F
authors.lastname: Fezza
authors.forename: Sergio
authors.initials: S
authors.lastname: Oddi
authors.forename: Monia
authors.initials: M
authors.lastname: Di Tommaso
authors.forename: Chiara
authors.initials: C
authors.lastname: De Simone
authors.forename: Cinzia
authors.initials: C
authors.lastname: Rapino
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Pasquariello
authors.forename: Enrico
authors.initials: E
authors.lastname: Dainese
authors.forename: Alessandro
authors.initials: A
authors.lastname: Finazzi-Agrò
authors.forename: Mauro
authors.initials: M
authors.lastname: Maccarrone
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D001710:Biotin / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D063388:Endocannabinoids; D005455:Fluorescent Antibody Technique; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D051379:Mice; D008856:Microscopy, Fluorescence; D053284:Polyunsaturated Alkamides / Q000378:metabolism*; D043882:Receptors, Cannabinoid / Q000378:metabolism
chemical_list: D001095:Arachidonic Acids; D063388:Endocannabinoids; D053284:Polyunsaturated Alkamides; D043882:Receptors, Cannabinoid; D001710:Biotin; C078814:anandamide