Alpha adrenergic receptors in the rabbit bladder base smooth muscle: alpha-1 adrenergic receptors mediate contractile...

Alpha adrenergic receptors in rabbit bladder base smooth muscle were investigated by in vitro responses of smooth muscle strips to exogenous alpha agonist stimulation and in radioligand binding assays. Norepinephrine and phenylephrine caused significantly greater maximal contractile responses than did clonidine. Also, the contractile response was only inhibited weakly by the alpha-2 selective antagonist yohimbine but was potently inhibited by alpha-1 selective antagonists prazosin and BE2254, suggesting that the response is mediated predominantly by alpha-1 adrenergic receptors. The alpha-1 selective antagonist [125I]BE2254 was used to specifically label a single class of binding sites with a dissociation constant of 131.0 +/- 5.9 pM and a maximal binding capacity of 17.6 +/- 1.9 fmol/mg of protein. Catecholamines compete for [125I]BE2254 binding stereospecifically and with the characteristic alpha adrenergic potency series of (-)-epinephrine greater than (-)-norepinephrine much greater than (-)-isoproterenol. The alpha-1 selective antagonist prazosin (Kd = 2.4 nM) is much more potent in competing for [125I]BE2254 binding than is the alpha-2 selective antagonist yohimbine (Kd = 2900 nM). Also, this dissociation constant of prazosin of [125I]BE2254 binding for bladder base smooth muscle membranes was similar to prazosin's pA2 value of 8.23 to 8.58 in the contraction experiments. The results suggest that alpha-1 rather than alpha-2 receptors predominantly mediate catecholamine-induced contraction in the rabbit bladder base. Also, these receptor sites can be measured directly with the specific antagonist radioligand, [125I]BE2254.