Abstract
A Chinese hamster ovary (CHO) cell line expressing the firefly luciferase gene under the control of six cAMP response elements (CREs) was stably transfected with the long form of the rat D 2 dopamine receptor. Saturation binding analysis using [ 3H]spiperone showed that the receptor was expressed at low levels ( B max = 96.5 ± 15.8 fmol/mg), but with an affinity characteristic of the D 2 receptor ( K d = 21.5 ± 3.7 pM). Luciferase expression in this cell line was modified in a dose dependent manner with dopamine receptor agonists ( N-propylapomorphine > apomorphine > quinpirole > dopamine) and antagonists (spiperone > (+)-butaclamol > D0710 > (−)-sulpiride > tiapride > remoxipride), according to their rank order of potency in binding and cAMP accumulation studies. Dopamine-mediated inhibition of forskolin-stimulated luciferase expression was pertussis toxin sensitive. This demonstrated the efficiency of the luciferase reporter gene assay for the functional testing of D 2 dopamine receptors, which are negatively coupled to the adenylyl cyclase signaling pathway, when heterogously expressed at low levels in CHO cells.
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