Agonist interaction with alkylation-sensitive and -resistant alpha-1 adrenoceptor subtypes

The interaction of agonists with alpha-1 receptor subtypes sensitive and resistant to alkylation by a prazosin analog [1-(4-amino-6,7-dimethoxy-2-quinazolinyl)-4-(2-bicyclo[2.2.2]octa-2,5- diene-z-carbonyl)-piperazine; SZL-49] has been examined. In rat aortic rings, SZL-49 (0.1-10 nM) shifted the dose-response curves for norepinephrine and phenylephrine to the right. The curves were biphasic, consisting of high and low affinity components. At greater than 10 nM, the curves became monophasic. After SZL-49 treatment, the response to norepinephrine was partially antagonized by diltiazem. Chlorethylclonidine (1-100 microM) also produced biphasic dose-response curves. Phenylephrine bound to high and low affinity sites labeled by [3H]prazosin, and the high affinity site was eliminated by SZL-49. SZL-49 (i.p.) shifted the pressor dose-response curve for phenylephrine to the right but did not decrease the maximal response. Chlorethylclonidine was much less potent than SZL-49 at shifting the pressor dose-response curve. Pertussis toxin, 50 micrograms/kg i.v., shifted the phenylephrine pressor dose-response curve in control and SZL-49-treated animals. SZL-49 inhibited norepinephrine-induced inositol phosphate formation, whereas chlorethylclonidine had no effect on inositol phosphate formation. These data show: 1) both in vitro and in vivo, alpha-1 receptor subtypes sensitive and resistant to alkylation by SZL-49 can mediate the full response of agonists; 2) these subtypes exhibit high and low affinity for agonists; 3) responses mediated by either subtype are partially dependent on calcium channel activity and a pertussis toxin-sensitive G-protein; 4) the SZL-49 sensitive site is able to enhance the formation of inositol phosphates.