Premium
Oleamide is a selective endogenous agonist of rat and human CB 1 cannabinoid receptors
Author(s) -
Leggett James D,
Aspley S,
Beckett S R G,
D'Antona A M,
Kendall D A,
Kendall D A
Publication year - 2004
Publication title -
british journal of pharmacology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.432
H-Index - 211
eISSN - 1476-5381
pISSN - 0007-1188
DOI - 10.1038/sj.bjp.0705607
Subject(s) - gtpgammas , agonist , cannabinoid , chemistry , cannabinoid receptor , cannabinoid receptor antagonist , stereochemistry , receptor , biochemistry
The ability of the endogenous fatty acid amide, cis ‐oleamide (ODA), to bind to and activate cannabinoid CB 1 and CB 2 receptors was investigated. ODA competitively inhibited binding of the nonselective cannabinoid agonist [ 3 H]CP55,940 and the selective CB 1 antagonist [ 3 H]SR141716A to rat whole‐brain membranes with K i values of 1.14 μ M (0.52–2.53 μ M , Hill slope=0.80, n =6) and 2.63 μ M (0.62–11.20 μ M , Hill slope=0.92, n =4), respectively. AEA inhibited [ 3 H]CP55,940 binding in rat whole‐brain membranes with a K i of 428 n M (346–510 n M , Hill slope=−1.33, n =3). ODA competitively inhibited [ 3 H]CP55,940 binding in human CB 1 (hCB 1 ) cell membranes with a K i value of 8.13 μ M (4.97–13.32 μ M , n =2). In human CB 2 transfected (hCB 2 ) HEK‐293T cell membranes, 100 μ M ODA produced only a partial (42.5±7%) inhibition of [ 3 H]CP55,940 binding. ODA stimulated [ 35 S]GTP γ S binding in a concentration‐dependent manner (EC 50 =1.64 μ M (0.29–9.32 μ M ), R 2 =0.99, n =4–9), with maximal stimulation of 188±9% of basal at 100 μ M . AEA stimulated [ 35 S]GTP γ S binding with an EC 50 of 10.43 μ M (4.45–24.42 μ M , R 2 =1.00, n =3, 195±4% of basal at 300 μ M ). Trans ‐oleamide ( trans‐ ODA) failed to significantly stimulate [ 35 S]GTP γ S binding at concentrations up to 100 μ M . ODA (10 μ M )‐stimulated [ 35 S]GTP γ S binding was reversed by the selective CB 1 antagonist SR141716A (IC 50 =2.11 n M (0.32–13.77 n M ), R 2 =1.00, n =6). The anatomical distribution of ODA‐stimulated [ 35 S]GTP γ S binding in rat brain sections was indistinguishable from that of HU210. Increases of similar magnitude were observed due to both agonists in the striatum, cortex, hippocampus and cerebellum. ODA (10 μ M ) significantly inhibited forskolin‐stimulated cyclic AMP (cAMP) accumulation in mouse neuroblastoma N1E 115 cells ( P =0.02, n =11). ODA‐mediated inhibition was completely reversed by 1 μ M SR141716A ( P <0.001, n =11) and was also reversed by pretreatment with 300 ng ml −1 pertussis toxin ( P <0.001, n =6). These data demonstrate that ODA is a full cannabinoid CB 1 receptor agonist. Therefore, in addition to allosteric modulation of other receptors and possible entourage effects due to fatty acid amide hydrolase inhibition, the effects of ODA may be mediated directly via the CB 1 receptor.British Journal of Pharmacology (2004) 141 , 253–262. doi: 10.1038/sj.bjp.0705607