Characterization of biotin-anandamide, a novel tool for the visualization of anandamide accumulation |
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Authors: | Fezza Filomena Oddi Sergio Di Tommaso Monia De Simone Chiara Rapino Cinzia Pasquariello Nicoletta Dainese Enrico Finazzi-Agrò Alessandro Maccarrone Mauro |
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Affiliation: | European Center for Brain Research/Istituto di Ricovero e Cura a Carattere Scientifico S. Lucia Foundation, Rome, Italy. |
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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. |
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Keywords: | endocannabinoids immunofluorescence keratinocyte transport metabolism skin |
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