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• W2067114461 abstract "The purpose of this study was to test whether screening at dopamine receptors performed with a recently described functional assay for G-protein coupled receptors (GPCRs) provides data that correlate significantly with radioligand binding data in the literature, thus possibly allowing researchers to replace radioligand binding with nonradioactive functional screening. Human dopamine receptors hD1 and hD2L (representing Gs [hD1] or G [hD2L] coupled GPCRs) were recombinantly expressed in human embryonic kidney (HEK293) cells. Cells were loaded with Oregon Green 488 BAPTA-1/AM and evenly distributed in 384 well plates. Seventeen test compounds were screened for agonistic activity by injection into the cell suspension and monitoringH of intracellular Ca2+ with a fluorescence microplate reader. Then, standard agonists (100nM SKF38393 for hD1, 30nM quinpirole for hD2L) were injected into wells preincubated with test compounds (screening for antagonism). Injection of various agonists resulted in a concentration-dependent increase in fluorescence. Further, preincubation of antagonists with dopamine receptor expressing cells inhibits concentration-dependent the agonist-induced increase in fluorescence. Calculated apparent functional Ki values correlate with radioligand binding data in the literature (r2=0.7796 for D1, r2=0.7743 for D2). The correlation between apparent functional K values and radioligand binding data for the 17 tested compounds suggests that screening of test compounds at dopamine receptors with the functional Ca2+ assay can replace radioligand binding studies. Furthermore, besides apparent K values, information about agonistic or antagonistic properties of a test compound can be obtained with the functional Ca2+ assay." @default.
• W2067114461 created "2016-06-24" @default.
• W2067114461 creator A5072810019 @default.
• W2067114461 date "2002-12-01" @default.
• W2067114461 modified "2023-10-16" @default.
• W2067114461 title "Quantitative comparison of functional screening by measuring intracellular Ca2+ with radioligand binding at recombinant human dopamine receptors" @default.
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• W2067114461 doi "https://doi.org/10.1208/ps040431" @default.
• W2067114461 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/2751320" @default.
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• W2067114461 hasPublicationYear "2002" @default.
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