title: Interactions of agonists with D-2 dopamine receptors: evidence for a single receptor population existing in multiple agonist affinity-states in rat striatal membranes.
abstract: Several lines of evidence previously indicated that [3H]spiroperidol (SPIRO) or [3H]domperidone (DOMP) might label heterogeneous populations of striatal dopamine receptors in radioligand binding studies. We have examined this possibility in rat striatum using computerized non-linear curve fitting and a method to block the unwanted [3H]SPIRO binding to S-2 serotonergic receptors. In the absence of a serotonergic antagonist, [3H]SPIRO saturation data produce curved Scatchard plots which are best computer fit by assuming the presence of two sites of different [3H]SPIRO affinities. In the presence of appropriate concentrations of ketanserin to block S-2 serotonergic binding, Scatchard plots are linear, with data modeling best to a single population of homogeneous binding sites. The D-2 dopamine receptor Bmax and [3H]SPIRO KD determined in this fashion are indistinguishable from that obtained for the higher affinity binding site by computer analysis of data obtained in the absence of ketanserin. [3H]DOMP produced indistinguishable values for D-2 receptor Bmax as well. Competitions by (-)sulpiride, metoclopramide, and DOMP for [3H]SPIRO binding sites in the presence of ketanserin are steep (Hill slope approximately 1), demonstrating that the previously observed heterogeneity of these sites is due entirely to serotonergic [3H]SPIRO binding. In contrast, agonist/3H-antagonist competition curves in the presence of ketanserin are best computer fit by assuming two independent receptor sites of high (RH) and low (RL) agonist affinity. With the addition of 5'-guanylylimidodiphosphate [Gpp(NH)p] computer analyses of agonist/3H-antagonist competition curves show an increased ratio RL/RH concomitant with apparent decreases in agonist affinities for both sites. Under some conditions, some agonist/3H-antagonist competition curves are best fit by a single site model in which agonist affinity is indistinguishable from the agonist's affinity at RL determined in the absence of Gpp(NH)p. These data are consistent with the presence of a single dopaminergic 3H-butyrophenone binding site, representing the D-2 receptor, which exists in two interconverting states differing in agonist affinity.
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002090:Butyrophenones / Q000378:metabolism*; D003342:Corpus Striatum / Q000032:analysis*; D004294:Domperidone / Q000378:metabolism*; D018492:Dopamine Antagonists; D006150:Guanine Nucleotides / Q000494:pharmacology; D007650:Ketanserin; D008787:Metoclopramide / Q000494:pharmacology; D010880:Piperidines / Q000494:pharmacology; D011869:Radioligand Assay; D051381:Rats; D011954:Receptors, Dopamine / Q000032:analysis* / Q000187:drug effects / Q000378:metabolism; D012964:Sodium / Q000494:pharmacology; D013134:Spiperone / Q000378:metabolism*; D013696:Temperature
chemical_list: D002090:Butyrophenones; D018492:Dopamine Antagonists; D006150:Guanine Nucleotides; D010880:Piperidines; D011954:Receptors, Dopamine; D013134:Spiperone; D004294:Domperidone; D007650:Ketanserin; D012964:Sodium; D008787:Metoclopramide