Functional analysis of the D-2L dopamine receptor expressed in a cAMP-responsive luciferase reporter cell line

George, Samantha E. and Bungay, Peter J. and Naylor, Louise H. (1998) Functional analysis of the D-2L dopamine receptor expressed in a cAMP-responsive luciferase reporter cell line. Biochemical Pharmacology, 56 (1). pp. 25-30. ISSN 0006-2952. (The full text of this publication is not available from this repository)

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Official URL
http://dx.doi.org/10.1016/S0006-2952(98)00014-8

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 [H-3]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 > (+) butaclarnol > 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.

Item Type: Article
Subjects: R Medicine > RS Pharmacy and materia medica
Divisions: Faculties > Science Technology and Medical Studies > School of Biosciences
Depositing User: R.F. Xu
Date Deposited: 09 Apr 1914 18:18
Last Modified: 18 Jun 2014 11:20
Resource URI: http://kar.kent.ac.uk/id/eprint/17760 (The current URI for this page, for reference purposes)
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