TY - JOUR
T1 - Characterization of biotin-anandamide, a novel tool for the visualization of anandamide accumulation
AU - Fezza, Filomena
AU - Oddi, Sergio
AU - Di Tommaso, Monia
AU - De Simone, Chiara
AU - Rapino, Cinzia
AU - Pasquariello, Nicoletta
AU - Dainese, Enrico
AU - Finazzi-Agrò, Alessandro
AU - Maccarrone, Mauro
PY - 2008/6/1
Y1 - 2008/6/1
N2 - 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.
AB - 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.
KW - Endocannabinoids
KW - Immunofluorescence
KW - Keratinocyte
KW - Metabolism
KW - Skin
KW - Transport
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U2 - 10.1194/jlr.M700486-JLR200
DO - 10.1194/jlr.M700486-JLR200
M3 - Article
C2 - 18316795
AN - SCOPUS:48549091727
VL - 49
SP - 1216
EP - 1223
JO - Journal of Lipid Research
JF - Journal of Lipid Research
SN - 0022-2275
IS - 6
ER -