Functional characterization of the human dopamine D4.2 receptor using vaccinia virus as an expression system

Claudia Bouvier, James R. Bunzow, Hong Chang Guan, Anja Unteutsch, Olivier Civelli, David K. Grandy, Hubert H.M. Van Tol

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Recombinant vaccinia viruses harboring the human dopamine D4 receptor cDNA containing two 48 base pair-repeats (D4.2) or the rat dopamine D2 short (D2s) receptor cDNA were used to infect rat-1 fibroblasts. Heterologous expression of both dopamine receptors was demonstrated in binding assays. The affinity constants of these receptors were consistent with values previously reported, including D4.2's higher affinity for the antipsychotic clozapine and raclopride's selectivity for D2 receptors. In the presence of 200 μM 5′-guanylyl- imidodiphosphate (Gpp[NH]p) both receptors exhibited reduced affinities for dopamine. Furthermore, when rat-1 cells were infected with the D2s or the D4.2 recombinant vaccinia viruses and exposed to dopamine agonists, the inhibition of adenylyl cyclase activity was prevented in pertussis toxin-treated cells. This study demonstrates the utility of recombinant receptor-vaccinia viruses in studies of expression, pharmacology and functional coupling of inhibitory G protein-coupled receptors.

Original languageEnglish (US)
Pages (from-to)11-17
Number of pages7
JournalEuropean Journal of Pharmacology: Molecular Pharmacology
Volume290
Issue number1
DOIs
StatePublished - Jun 23 1995
Externally publishedYes

Keywords

  • (Functional expression)
  • Adenylyl cyclase
  • Dopamine D receptor
  • Vaccinia virus

ASJC Scopus subject areas

  • Pharmacology

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