PHARMACOLOGICAL ANALYSIS OF DOPAMINE STIMULATION OF [S-35] GTP-GAMMA-S BINDING VIA HUMAN D-2SHORT AND D-2LONG DOPAMINE-RECEPTORS EXPRESSED IN RECOMBINANT CELLS
B. Gardner et al., PHARMACOLOGICAL ANALYSIS OF DOPAMINE STIMULATION OF [S-35] GTP-GAMMA-S BINDING VIA HUMAN D-2SHORT AND D-2LONG DOPAMINE-RECEPTORS EXPRESSED IN RECOMBINANT CELLS, British Journal of Pharmacology, 118(6), 1996, pp. 1544-1550
1 The activation of G-proteins by agonist-occupied D-2 or D-3 dopamine
receptors in membranes from recombinant cells expressing the cloned r
eceptors has been analysed by a [S-35]-guanosine 5'-[gamma-thio] triph
osphate ([S-35]-GTP gamma S) binding assay. 2 The rate of [S-35]-GTP g
amma S binding was increased by dopamine in a dose-dependent manner in
membranes from CHO cells stably expressing either the D-2short or D-2
long dopamine receptor. 3 The dopamine-induced stimulation of [S-35]-G
TP gamma S binding could be inhibited by a range of antagonists. Affin
ities for antagonists derived from the inhibition of the dopamine stim
ulation of [S-35]- GTP gamma S binding correlated very well with affin
ities derived from radioligand binding studies. 4 When the maximum [S-
35]-GTP gamma S binding responses stimulated by dopamine acting at dif
ferent receptor subtypes were compared, there was a tendency for the s
timulation via the D-2short receptor to be greater than via the D-2lon
g receptor and for the stimulation via the D-3 dopamine receptor to be
less than for either D-2 receptor. These differences in maximal respo
nse were also seen when the inhibitory effects of dopamine on adenylyl
cyclase via the three receptor subtypes were compared. 5 The stimulat
ion of [S-35]-GTP gamma S binding by dopamine in membranes from recomb
inant cells therefore provides an excellent system for studying the mo
lecular nature of agonism and the receptor/G-protein interactions for
these receptors.