Submitted on October 26, 2007
Revised on March 3, 2008
Accepted on March 3, 2008
Characterization of biotin-anandamide, a novel tool for the visualization of anandamide accumulation
Filomena Fezza, Sergio Oddi, Monia Di Tommaso, Chiara De Simone, Cinzia Rapino, Nicoletta Pasquariello, Enrico Dainese, Alessandro Finazzi-Agrò, and Mauro Maccarrone
Department of Biomedical Sciences, University of Teramo, Teramo
Corresponding Author: mmaccarrone{at}unite.it
Anandamide (arachidonoylethanolamide, AEA) acts as endogenous agonist of both cannabinoid and vanilloid receptors. During the last two decades, its metabolic pathways and biological activity have been extensively investigated and relatively well-characterized. In contrast, the effective nature and mechanism of AEA transport remain at present a controversial and still unsolved issue. Here we report the characterization of a biotinylated analogue of AEA (b-AEA), that has the same lipophylicity 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 nor interfere with the other components of the endocannabinoid system, i.e. type-1 and type-2 cannabinoid receptors, vanilloid receptor, AEA synthetase (NAPE-PLD) or AEA hydrolase (FAAH). Taken together, our data suggest that b-AEA could be a very useful probe for visualizing the accumulation and intracellular distribution of this endocannabinoid.
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