title: Characterization of alpha-1 adrenergic receptors in the heart using [3H]WB4101: effect of 6-hydroxydopamine treatment.
abstract: To identify and characterize the cardiac alpha-adrenoceptors, a radioreceptor binding assay using the potent alpha adrenergic antagonist, [3H]WB4101 was performed in rat hearts. Specific [3H]WB4101 binding to rat left ventricular homogenates was saturable, reversible and of high affinity (Kd = 0.18 nM) with a Bmax of 2.57 fmol/mg of tissue (27.7 fmol/mg of protein). Adrenergic agonists competed for specific [3H]WB4101 binding in the order: (-)-epinephrine > (-)-norepinephrine greater than (-)-isoproterenol. Stereospecificity of the [3H]WB4101 binding sites was also demonstrated with (-)-epinephrine, (Ki = 90) nM being 270 times as potent as (+)-epinephrine, (K1 = 24 microM). Adrenergic antagonists competed for the binding in the order: WB4101 = prazosin greater than yohimbine greater than (-)-propranolol. WB4101 and prazosin exhibited a markedly greater (2000 times) affinity for [3H]WB4101 binding sites than yohimbine. The affinities (pKi) of alpha agonists and antagonists for [3H]WB4101 binding sites in the rat heart closely correlated with their pharmacological potencies in the heart. Scatchard analysis for [3H]WB4101 binding, performed in five regions from control and 6-hydroxydopamine-treated rat hearts, revealed specific [3H]WB4101 binding (Bmax) significantly greater in the ventricles and intraventricular septae than in atria. At 1 week after 6-hydroxydopamine treatment, there was a significant increase (40%) in the Bmax for [3H]WB4101 binding to ventricles and intraventricular septae without a change in Kd. We conclude: 1) [3H]Wb4101 selectively labels postsynaptic alpha-1 adrenoceptors in the rat heart; 2) there is a definite regional variation for cardiac alpha-1 adrenoceptors; and 3) 6-hydroxydopamine treatment caused a significant increase in the density of alpha-1 adrenoceptors in ventricles and intraventricular septae, compatible with a postsynaptic localization of the [3H]WB4101 binding site.
mesh_terms: D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology*; D000818:Animals; D001665:Binding Sites; D004146:Dioxanes / Q000494:pharmacology*; D004147:Dioxins / Q000494:pharmacology*; D006321:Heart / Q000294:innervation*; D006892:Hydroxydopamines / Q000494:pharmacology*; D008297:Male; D051381:Rats; D011941:Receptors, Adrenergic / Q000032:analysis*; D011942:Receptors, Adrenergic, alpha / Q000032:analysis*; D013564:Sympathetic Nervous System / Q000187:drug effects*
chemical_list: D000317:Adrenergic alpha-Antagonists; D004146:Dioxanes; D004147:Dioxins; D006892:Hydroxydopamines; D011941:Receptors, Adrenergic; D011942:Receptors, Adrenergic, alpha; C010654:(2-(2',6'-dimethoxy)phenoxyethylamino)methylbenzo-1,4-dioxane