Antagonist-D2S-dopamine receptor interactions in intact recombinant Chinese hamster ovary cells [corrected].

D(2)-type dopamine receptors are major recognition sites for antipsychotic drugs. There are two splice variants: D(2S) and D(2L) with an additional 29 amino acid sequence in the third intracellular loop. Only little comparative information is hitherto available about their pharmacological properties and none of these studies dealt with intact cell systems. This prompted us to investigate the binding properties of [(3)H]-raclopride, a hydrophilic benzamide, and [(3)H]-spiperone, a highly hydrophobic butyrophenone, to intact CHO cells expressing recombinant human D(2L)-receptors. Presently, we have repeated and extended this experimental approach to the human D(2S)-receptors in the same cell system. Except for a slower dissociation of [(3)H]-spiperone from D(2S), the binding properties of these and other antagonists were not significantly different for both isoforms (P > 0.05). The very slow dissociation of the atypical antipsychotic clozapine was surprising in light of its low affinity. Two experiments pointed out the existence of non-competitive interactions between raclopride and spiperone for D(2S) as well as D(2L) (A. Packeu, J. P. De Backer & G. Vauquelin, in preparation). Alongside the different physicochemical properties of these ligands, this finding fits with a model wherein the hydrophilic raclopride approaches the D(2L)-receptor from the aqueous phase, while the hydrophobic spiperone approaches the receptor by lateral diffusion between the membrane lipids. These different modes of approach could imply the existence of topologically distinct ligand binding sites at D(2)-receptors.