gw9662 and anandamide

We investigated the effects of anandamide on peroxisome proliferator-activated receptor gamma (PPARgamma) activity. In two different transactivation systems using either full-length or only the ligand binding domain of PPARgamma, we showed that anandamide, but not palmitoylethanolamide induced transcriptional activation of PPARgamma in a dose dependent manner with an EC50 of 8 microM. In addition, competition binding experiments showed that anandamide but not palmitoylethanolamide binds directly to PPAR-ligand binding domain. We also found that anandamide treatment induced 3T3-L1 fibroblast differentiation into adipocytes. Indeed, anandamide induced triglyceride droplet accumulation and the expression of PPARgamma responsive genes such as CCAAT enhancer binding protein alpha (C-EBPalpha), aP2, PerilipinA and Acrp30. Furthermore, the PPARgamma antagonist (GW9662) inhibited the anandamide-induced 3T3-L1 differentiation confirming that this is a PPARgamma-mediated process. Altogether, these data indicate that anandamide binds PPARgamma and induces cellular PPARgamma signaling.

Topics: 3T3-L1 Cells; Adipocytes; Adiponectin; Anilides; Animals; Arachidonic Acids; Binding Sites; Binding, Competitive; Blotting, Western; Cannabinoid Receptor Modulators; Carrier Proteins; CCAAT-Enhancer-Binding Protein-alpha; Cell Differentiation; Cell Line; Chlorocebus aethiops; Chromans; COS Cells; Dose-Response Relationship, Drug; Endocannabinoids; Fibroblasts; Gene Expression; HeLa Cells; Humans; Intercellular Signaling Peptides and Proteins; Luciferases; Mice; Perilipin-1; Phosphoproteins; Pioglitazone; Plasmids; Polyunsaturated Alkamides; PPAR gamma; Recombinant Fusion Proteins; Reverse Transcriptase Polymerase Chain Reaction; Thiazolidinediones; Transcriptional Activation; Transfection; Troglitazone