title: Endogenous cannabinoid anandamide inhibits nicotinic acetylcholine receptor function in mouse thalamic synaptosomes.
abstract: The effects of the endogenous cannabinoid anandamide [arachidonylethanolamide (AEA)] on the function of nicotinic acetylcholine receptor (nAChR) were investigated using the 86Rb+ efflux assay in thalamic synaptosomes. AEA reversibly inhibited 86Rb+ efflux induced by 300 microM ACh with an IC50 value of 0.9 +/- 2 microM. Pre-treatment with the cannabinoid (CB1) receptor antagonist SR141716A (1 microM), the CB2 receptor antagonist SR144528 (1 microM), or pertussis toxin (0.2 mg/mL) did not alter the inhibitory effects of AEA, suggesting that known CB receptors are not involved in AEA inhibition of nAChRs. AEA inhibition of 86Rb+ efflux was not reversed by increasing acetylcholine (ACh) concentrations. In radioligand binding studies, the specific binding of [3H]-nicotine was not altered in the presence of AEA, indicating that AEA inhibits the function of nAChR in a non-competitive manner. Neither the amidohydrolase inhibitor phenylmethylsulfonyl fluoride (0.2 mM) nor the cyclooxygenase inhibitor, indomethacin, (5 microM) affected AEA inhibition of nAChRs, suggesting that the effect of AEA is not mediated by its metabolic products. Importantly, the extent of AEA inhibition of 86Rb+ efflux was significantly attenuated by the absence of 1% fatty acid free bovine serum albumin pre-treatment, supporting previous findings that fatty acid-like compounds modulate the activity of nAChRs. Collectively, the results indicate that AEA inhibits the function of nAChRs in thalamic synaptosomes via a CB-independent mechanism and that the background activity of these receptors is affected by fatty acids and AEA.
authors.affiliation: Neuroscience Discovery, Martek Biosciences, Boulder, Colorado, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Butt
authors.forename: Alp
authors.initials: A
authors.lastname: Alptekin
authors.forename: Toni
authors.initials: T
authors.lastname: Shippenberg
authors.forename: Murat
authors.initials: M
authors.lastname: Oz
mesh_terms: D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism / Q000502:physiology*; D063385:Cannabinoid Receptor Modulators / Q000378:metabolism / Q000502:physiology*; D063388:Endocannabinoids; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018733:Nicotinic Antagonists / Q000378:metabolism*; D053284:Polyunsaturated Alkamides / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D011978:Receptors, Nicotinic / Q000502:physiology*; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D013788:Thalamus / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*
chemical_list: D001095:Arachidonic Acids; D063385:Cannabinoid Receptor Modulators; D063388:Endocannabinoids; D018733:Nicotinic Antagonists; D053284:Polyunsaturated Alkamides; D011978:Receptors, Nicotinic; C078814:anandamide