CHARACTERIZATION OF ALPHA-1-ADRENERGIC RECEPTOR SUBTYPES IN RAT-BRAIN - A REEVALUATION OF [H-3] WB4104 AND [H-3] PRAZOSIN BINDING

Abstract

[3H]Prazosin and [3H]WB4101 [2-(2,6-dimethoxyphenoxyethyl)aminomethyl-1,4 benzodioxane] have both been proposed to label .alpha.1-adrenergic receptors in the rat central nervous system. As many discrepancies between the binding of these two ligands have arisen, we conducted these studies in order to reevaluate their binding characteristics and resolve the similarities and differences in the pharmacological characteristics of their respective binding sites. [3H]Prazosin binding is characterized by a monophasic saturation isotherm. Prazosin, indoramine, and dihydroergocryptine competitions with [3H]prazosin are steep and monophasic, and model best to a single binding site. In contrast, phentolamine and WB4101 competition curves are shallow in rat cortex, exhibiting Hill coefficients significantly less than 1.0, and model to two binding sites of approximately equal proportions. The higher and lower affinity components are defined as .alpha.1A and .alpha.1B, respectively. [3H]WB4101 also labels two binding sites in rat cortex and hippocampus with picomolar and nanomolar affinity, respectively. However, the nanomolar binding site is serotonergic and not adrenergic. The picomolar site (Kd = 150 pM) has characteristics of an .alpha.1-receptor binding site:prazosin, WB4101, and phentolamine affinities for this [3H]WB4101 binding site correlate with their affinities for the highest affinity component (.alpha.1A) of [3H]prazosin binding. In addition, the Bmax of this [3H]WB4101-labeled site is equal to one-half of the total [3H]prazosin Bmax. Agonist competitions with [3H]prazosin binding are multiphasic with pseudo-Hill slopes less than 1.0 and with a rank order of affinity of epinephrine > norepinephrine > phenylephrine. When binding to the .alpha.1A component is blocked by a 30 mM phentolamine mask, the same rank order of agonist affinities is preserved. Although the affinities of epinephrine and norepinephrine at the two subtypes are identical, phenylephrine is weaker at the .alpha.1B site. The ratio of the potency of phentolamine versus prazosin is about 4 at the .alpha.1A component but about 80 at the .alpha.1B binding site. We discuss these data in relation to the reported potencies of these antagonists in blocking .alpha.1-receptor-mediated responses which may correlate with our designation of .alpha.1A or .alpha.1B binding sites.