title: Method to denude rat aortic endothelium with saponin for phosphoinositide analysis in vascular smooth muscle.
abstract: Arterial endothelium plays an essential role in modulating vascular tone through the production of vasoactive biogenic substances. To determine the effect of endothelial cell function on vascular smooth muscle receptor-effector coupling, a simple chemical techniques was devised to denude rat aortic endothelium while receptor function was preserved. Agonist-stimulated phosphatidylinositol turnover (PI) and vascular contractility were studied in saponin-treated aortic rings. Uniform removal of the aortic endothelial cell layer was readily verified with the use of a modified silver stain with light microscopy. Agonist-mediated PI turnover was statistically decreased in saponin-exposed aortic rings relative to control aorta. Increased contractile sensitivity in denuded aorta was also observed with norepinephrine (NE), phenylephrine (PE), and serotonin (5-HT). Thus, brief exposure to saponin is an effective chemical skinning technique that preserves vascular function.
authors.affiliation: Department of Internal Medicine, Texas Tech University Health Sciences Center, Lubbock 79430.
authors.forename: E
authors.initials: E
authors.lastname: Legan
authors.forename: J A
authors.initials: JA
authors.lastname: Sisson
mesh_terms: D000818:Animals; D001011:Aorta; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D008297:Male; D008854:Microscopy, Electron; D009638:Norepinephrine / Q000494:pharmacology; D010656:Phenylephrine / Q000494:pharmacology; D010716:Phosphatidylinositols / Q000032:analysis / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012503:Saponins / Q000494:pharmacology*; D012701:Serotonin / Q000494:pharmacology; D013048:Specimen Handling / Q000379:methods*; D014661:Vasoconstriction
chemical_list: D010716:Phosphatidylinositols; D012503:Saponins; D010656:Phenylephrine; D012701:Serotonin; D009638:Norepinephrine