EXPRESSION AND PHARMACOLOGICAL CHARACTERIZATION OF THE HUMAN D-3 DOPAMINE-RECEPTOR

Binding of dopamine receptor ligands to human D-2 and D-3 receptors wa s characterized in Chinese hamster ovary (CHO) cells using the dopamin e D-2 receptor antagonist [I-125] iodosulpiride. Only limited binding selectivity, was observed for known dopamine D-2 receptor antagonists from a variety of chemical classes, which included haloperidol, chlorp romazine, sulpiride, pimozide and cis flupenthixol. The most selective compound from this group were (+)butaclamol and domperidone which sho wed 5-fold D-3 selectivity. A number of high affinity dopamine recepto r agonists, including apomorphine and bromocriptine, also failed to de monstrate selectivity. In contrast, the natural ligand dopamine and th e efficacious synthetic agonists quinpirole, (+)4-propyl-9-hydroxynapt hoxazine (PHNO), -amino-6,7-dihydroxy-1,2,3,4-tetrahydronaphthalene (6 ,7-ADTN), 7-OH DPAT and N-0434 showed marked apparent human dopamine D -3 (hD(3)) receptor selectivity. In the aminotetralin series, this sel ectivity was observed preferentially with analogs of the 6,7-rotamer c ompared with compounds from the 5,6-rotamer series. Functional couplin g of the hD(3) receptor was investigated in a number of cell lines in which the hD(3) receptor was stably expressed, including CHO cells, th e neuroblastoma-glioma hybrid cell line NG108-15 and a rat 1 fibroblas t cell line. There was no evidence of functional coupling of the hD(3) receptor to adenylate cyclase, arachidonic acid release, phospholipas e C activation, K+ currents or calcium mobilization in any of the cell lines examined. Furthermore, guanine nucleotides failed to inhibit th e binding of [H-3] N-0437 to hD(3) receptors in any of the three cell lines. There may be a number of explanations for these results. These cell lines may not have the appropriate G-protein or secondary messeng er systems that are coupled to the hD(3) receptor in situ. Alternative ly, this receptor may couple by a mechanism that is as yet undefined. The finding that a wide range of structurally diverse human dopamine D -2 (hD(2)) receptor agonists have an apparent hD(3) selectivity may im ply that the hD(3) receptor exists predominantly in a high affinity st ate.