A comprehensive method for the quantitative determination of dopamine receptor subtypes.
We have demonstrated that three subtypes of dopamine receptors can be characterized using several radioligand binding techniques. Indirect binding assays in which several competing ligands were used to inhibit the binding of the nonselective radioligand spiroperidol resulted in shallow displacement curves with Hill coefficients less than 1. Nonlinear regression analysis of these curves also indicated that there were two subtypes of the D-2 receptor present in a ratio of approximately 3 to 1. Direct binding assays with [3H]alpha-flupenthixol showed that this radioligand nonselectively labeled D-2A, D-2B, and D-1 receptors. Inhibition of the binding of [3H]alpha-flupenthixol by spiroperidol revealed that spiroperidol had a much higher affinity for D-2A and D-2B receptors than for D-1 receptors. Masking D-2 receptors with nanomolar concentrations of spiroperidol permitted characterization of D-1 receptors with the radioligand [3H]alpha-flupenthixol. Indirect binding assays of D-1 receptors with numerous competing ligands resulted in steep displacement curves with Hill coefficients of 1. This is consistent with the existence of a single, homogeneous population of D-1 receptors.
D000818:Animals; D001667:Binding, Competitive; D003342:Corpus Striatum / Q000378:metabolism; D004298:Dopamine / Q000031:analogs & derivatives / Q000378:metabolism; D018492:Dopamine Antagonists; D005475:Flupenthixol / Q000378:metabolism; D019204:GTP-Binding Proteins; D006160:Guanosine Triphosphate / Q000494:pharmacology; D008433:Mathematics; D011869:Radioligand Assay / Q000379:methods*; D011956:Receptors, Cell Surface / Q000378:metabolism; D011954:Receptors, Dopamine / Q000032:analysis* / Q000378:metabolism; D011985:Receptors, Serotonin / Q000378:metabolism; D013134:Spiperone / Q000378:metabolism; D014316:Tritium
D018492:Dopamine Antagonists; D011956:Receptors, Cell Surface; D011954:Receptors, Dopamine; D011985:Receptors, Serotonin; D014316:Tritium; D013134:Spiperone; D006160:Guanosine Triphosphate; D019204:GTP-Binding Proteins; D005475:Flupenthixol; D004298:Dopamine