title: [3H]idazoxan binding to bovine adrenal medullary membranes: identification and pharmacological characterization of I2-imidazoline sites.
abstract: Bovine adrenal chromaffin cells, which have been shown to lack alpha 2-adrenoceptors, were used to investigate the pharmacological characteristics of [3H]idazoxan binding sites. The binding of [3H]idazoxan was very rapid, reversible, partly specific (as defined by cirazoline 0.1 mmol/l; 50% specific binding at [3H]idazoxan 10 nmol/l), saturable and of high affinity (KD 13nmol/l) and, hence, was compatible with the criteria for the identification of an imidazoline binding site (IBS). Since in competition experiments rauwolscine and (-)-adrenaline showed only negligible affinity for these adrenal medullary binding sites, the lack of alpha 2-adrenoceptors was confirmed. Histamine and amiloride also did not inhibit [3H]idazoxan binding or caused only negligible inhibition. In contrast, the specific binding of [3H]idazoxan was concentration-dependently inhibited by several imidazolines and guanidines with the following rank order of potency which conforms to the characteristics of the previously defined I2-IBS (in parentheses: Ki, nmol/l): idazoxan (4) = cirazoline (4) >> clonidine (272) = BDF 6143 (299; 4-chloro-2-(2-imidazoline-2-ylamino)-isoindoline) > BDF 6100 (563; 2-(2-imidazolin-2-yl-amino)-isoindoline) > or = BDF 7579 (868; (4-chloro-2-isoindolinyl)guanidine) > phentolamine (1424) = naphazoline (1451). Equilibrium [3H]idazoxan binding was reduced by K+ but not by Na+ or the non-hydrolysable GTP-analogue Gpp(NH)p (5'-guanylylimidodiphosphate; 100 mumol/l). In conclusion, membranes of the bovine adrenal medulla are endowed with non-adrenergic high-affinity [3H]idazoxan sites which exhibit the pharmacological properties of the amiloride-insensitive subtype of I2-IBS and probably are not coupled to a G-protein.
authors: Institute of Pharmacology and Toxicology, University of Bonn, Germany.
authors: G J
authors: GJ
authors: Molderings
authors: L
authors: L
authors: Kundt
authors: M
authors: M
authors: Göthert
mesh_terms: D000313:Adrenal Medulla / Q000378:metabolism* / Q000648:ultrastructure; D000317:Adrenergic alpha-Antagonists / Q000378:metabolism* / Q000493:pharmacokinetics; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002412:Cations; D002417:Cattle; D004146:Dioxanes / Q000378:metabolism* / Q000493:pharmacokinetics; D019791:Guanidine; D006146:Guanidines / Q000378:metabolism / Q000494:pharmacology; D006165:Guanylyl Imidodiphosphate / Q000494:pharmacology; D019329:Idazoxan; D007093:Imidazoles / Q000378:metabolism*; D007457:Iodine Radioisotopes; D007700:Kinetics; D008566:Membranes / Q000378:metabolism; D018341:Receptors, Adrenergic, alpha-2 / Q000378:metabolism; D014316:Tritium
chemical_list: D000317:Adrenergic alpha-Antagonists; D002412:Cations; D004146:Dioxanes; D006146:Guanidines; D007093:Imidazoles; D007457:Iodine Radioisotopes; D018341:Receptors, Adrenergic, alpha-2; D014316:Tritium; D006165:Guanylyl Imidodiphosphate; D019791:Guanidine; D019329:Idazoxan