PMID: 11918350Mar 29, 2002Paper

Patterns of cyclic AMP formation by coexpressed D1 and D2L dopamine receptors in HEK 293 cells

Receptors & Channels
D J Sakolsky, B Ashby

Abstract

Recent studies using immunofluorescence confocal microscopy (Aizman et al., Nature neuroscience (2000) 3, 226-230) present compelling evidence for colocalization of D1 and D2 dopamine receptors on neurons in the striatum and nucleus accumbens. To examine some of the biochemical consequences of colocalization we coexpressed the D1 and D2 dopamine receptors in HEK293 cells. Dopamine D1 and D2 receptors couple to stimulation and inhibition of adenylyl cyclase, respectively. In cells expressing only the D1 receptor, dopamine stimulated cAMP formation with an EC50 of 2.15 nM. In cells expressing only the D2L receptor, dopamine inhibited cAMP formation by 80% with an EC50 of 0.02 nM. The effect of dopamine on the D2L receptor was antagonized by the selective antagonist spiperone with an IC50 of 0.31 nM. In cells coexpressing both the D1 and D2L receptors, dopamine caused an increase in cAMP that was only 20% of that observed with the D1 receptor alone. In this case, increasing concentrations of spiperone caused a change in the dose-response curve from hyperbolic to bell-shaped as the concentration of spiperone was increased. Using pharmacological constants determined from studies on the individually expressed receptors, the curves ob...Continue Reading

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