Mono- and divalent cations modulate the affinities of brain D1 and D2 receptors for dopamine by a mechanism independent of receptor coupling to guanyl nucleotide binding proteins

Abstract

In order to clarify the question of whether the modulatory effects of cations on dopamine receptor affinities are brought about by shifts in the equilibrium of receptor - G protein - coupling, it was investigated whether mono- and divalent cations were still able to modulate rat striatal D1 and D2 receptor affinities after selective inactivation of the G-proteins linked to the two receptors. The Gs-protein coupled to the D1 receptor was eliminated by mild thermal inactivation, and the Gi- (or Go-) protein associated with the D2 receptor by alkylation with a low concentration of N-ethyl-maleimide. Incubation of striatal membranes at 60 degrees C completely abolished the specific binding of 3H-GTP. Both treatments resulted in an increase of the IC50-values for dopamine as a displacer of 3H-SCH 23390 from D1- and of 3H-spiperone from D2 receptors. Concomitantly, the formerly shallow D1 displacement curves became steeper, with their Hill coefficients increasing. This effect was less evident at D2 receptors. Guanosine triphosphate (GTP), which increased the IC50's of dopamine for both receptors approximately two-fold in control membranes, was without effect in pretreated samples, indicating an effective inactivation of the G-proteins. Na+ ions were still able to lower, and Ca2+ ions to increase the affinities of D1 and D2 receptors for dopamine after such inactivation of the respective G-proteins. It is concluded that the mechanism underlying the regulation of dopamine receptor affinities by mono- and divalent cations is independent of and superimposed upon the coupling of these receptors to guanyl nucleotide binding proteins.