Involvement of receptor reserve in D1 agonistic action of (-)-stepholidine in lesioned rats

(-)-Stepholidine (SPD) is a natural product. Previous studies had demonstrated that SPD displayed D1 agonism in unilaterally 6-hydroxydopamine (6-OHDA)-lesioned rats and D1 antagonism in reserpinized rats and normal rats. The aim of the present study was to explain this peculiar pharmacological action based on behavioral and biochemical experiments. In the unilaterally 6-OHDA-lesioned rats, SPD (4 mg/kg, s.c.) induced contralateral rotation as did apomorphine (APO), but the rotation response to SPD was 60% lower than that to APO (0.5 mg/kg, i.p.). Coadministration with APO (0.5 mg/kg, i.p.) and SPD (0.5 to 10 mg/kg, s.c.) produced a biphasic action curve. At low doses (0.5 or 1 mg/kg), SPD potentiated APO action; at high doses (4 or 10 mg/kg), however, SPD suppressed APO. In striatal homogenate of the unilaterally lesioned rats, SPD stimulated cyclic AMP (cAMP) formation and produced a maximal response comparable to that of dopamine (DA) in the denervated striatum, but 70% lower than that of DA in the intact striatum. Coadministration of 10 microM DA with various concentrations of SPD yielded different results, with a biphasic response in the intact side and a synergistic effect in the denervated side. Furthermore, based on the determination of receptor-mediated cAMP formation, the D1 receptor reserve was analyzed in both denervated and intact striatum by using the DA receptor inactivator N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ). The results showed that following EEDQ administration, the receptor density [revealed by [3H]R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-be nzazepine ([3H]SCH-23390) binding] and the agonist-stimulated adenylate cyclase (AC) activity (revealed by cAMP formation) were reduced concurrently. In the intact striatum, the reduction in SPD-stimulated AC activity paralleled the receptor loss, indicating the absence of receptor reserve, while in the denervated striatum the reduction in AC activity was less than the receptor loss, indicating a significant level of receptor reserve (estimated 16.4%). By comparison, receptor reserve for DA was 45.7 and 25.3% in the denervated and intact striatum, respectively, representing an 80% increase of receptor reserve. In conclusion, SPD is a D1 partial agonist, and receptor reserve permits SPD to display its D1 agonistic action in the unilaterally 6-OHDA-lesioned rats.