Anandamide (N-arachidonoylethanolamide; AEA) acts as an endogenous agonist ofboth cannabinoid and vanilloid receptors. During the last two decades, itsmetabolic pathways and biological activity have been investigated extensively andrelatively 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 hasthe same lipophilicity of the parent compound. In addition, by means ofbiochemical assays and fluorescence microscopy, we show that b-AEA is accumulatedinside the cells in a way superimposable on that of AEA. Conversely, b-AEA doesnot interact or interfere with the other components of the endocannabinoidsystem, such as type-1 and type-2 cannabinoid receptors, vanilloid receptor, AEA synthetase (N-acylphosphatidylethanolamine-hydrolyzing phospholipase D), or AEAhydrolase (fatty acid amide hydrolase). Together, our data suggest that b-AEAcould be a very useful probe for visualizing the accumulation and intracellulardistribution of this endocannabinoid.[...]
Characterization of biotin-anandamide, a novel tool for the visualization of anandamide accumulation
ODDI, Sergio;DI TOMMASO, MONIA;RAPINO, CINZIA;PASQUARIELLO, NICOLETTA;DAINESE, Enrico;MACCARRONE, Mauro
2008-01-01
Abstract
Anandamide (N-arachidonoylethanolamide; AEA) acts as an endogenous agonist ofboth cannabinoid and vanilloid receptors. During the last two decades, itsmetabolic pathways and biological activity have been investigated extensively andrelatively 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 hasthe same lipophilicity of the parent compound. In addition, by means ofbiochemical assays and fluorescence microscopy, we show that b-AEA is accumulatedinside the cells in a way superimposable on that of AEA. Conversely, b-AEA doesnot interact or interfere with the other components of the endocannabinoidsystem, such as type-1 and type-2 cannabinoid receptors, vanilloid receptor, AEA synthetase (N-acylphosphatidylethanolamine-hydrolyzing phospholipase D), or AEAhydrolase (fatty acid amide hydrolase). Together, our data suggest that b-AEAcould be a very useful probe for visualizing the accumulation and intracellulardistribution of this endocannabinoid.[...]I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
