title: U-46619 but not serotonin increases endocannabinoid content in middle cerebral artery: evidence for functional relevance.
abstract: Cerebral vascular smooth muscle cells express the CB(1) cannabinoid receptor, and CB(1) receptor agonists produce vasodilation of cerebral arteries. The purpose of this study was to determine whether vasoconstriction of rat middle cerebral artery (MCA) results in the local formation of endocannabinoids (eCBs), which, via activation of CB(1) receptors, oppose the vasoconstriction in a feedback manner. The thromboxane A(2) (TXA(2)) mimetic U-46619 significantly increased N-arachidonylethanolamine (AEA) and 2-arachidonylglycerol (2-AG) content of isolated MCA, whereas 5-hydroxytrypamine (5-HT) decreased AEA and 2-AG content. If eCBs play a feedback role in the regulation of MCA tone, then CB(1) receptor antagonists should enhance the constriction of MCA produced by U-46619 but not 5-HT. U-46619 caused concentration-dependent constrictions of endothelium-denuded MCA. Two CB(1) receptor antagonists SR-141716 and AM-251 decreased the EC(50) value for U-46619 to constrict endothelium-denuded MCA without affecting the maximal effect. A low concentration of CB(1) receptor agonist Win-55212-2 (30 nM) produced vasodilation of MCAs constricted with low but not saturating concentrations of U-46619. SR-141716 had no effect on the 5-HT concentration-contraction relationship. These data suggest that TXA(2) receptor activation increases MCA eCB content, which, via activation of CB(1) receptors, reduces the constriction produced by moderate concentrations of the TXA(2) agonist. Although 5-HT-induced vasoconstriction is reduced by exogenous CB(1) receptor agonist, activation of 5-HT receptors does not increase eCB content. These results suggest that MCA production of eCBs is not regulated by constriction per se but likely via a signaling pathway that is specific for TXA(2) receptors and not 5-HT receptors.
authors.affiliation: Medical College of Wisconsin, Dept. of Pharmacology and Toxicology, 8701 Watertown Plank Rd., Milwaukee, WI 53226-0509, USA. chillard@mcw.edu
authors.forename: David J
authors.initials: DJ
authors.lastname: Rademacher
authors.forename: Sachin
authors.initials: S
authors.lastname: Patel
authors.forename: W-S Vanessa
authors.initials: WS
authors.lastname: Ho
authors.forename: Amanda M
authors.initials: AM
authors.lastname: Savoie
authors.forename: Nancy J
authors.initials: NJ
authors.lastname: Rusch
authors.forename: Kathryn M
authors.initials: KM
authors.lastname: Gauthier
authors.forename: Cecilia J
authors.initials: CJ
authors.lastname: Hillard
mesh_terms: D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid / Q000494:pharmacology*; D000818:Animals; D020771:Anterior Cerebral Artery / Q000187:drug effects / Q000502:physiology*; D063385:Cannabinoid Receptor Modulators / Q000378:metabolism*; D004075:Diglycerides / Q000378:metabolism; D063388:Endocannabinoids; D066298:In Vitro Techniques; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000502:physiology; D010714:Phosphatidylethanolamines / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012701:Serotonin / Q000494:pharmacology*; D014662:Vasoconstrictor Agents / Q000494:pharmacology
chemical_list: D063385:Cannabinoid Receptor Modulators; D004075:Diglycerides; D063388:Endocannabinoids; C109143:N-arachidonylphosphatidylethanolamine; D010714:Phosphatidylethanolamines; D014662:Vasoconstrictor Agents; D012701:Serotonin; D019796:15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid; C035561:diarachidonyl diglyceride