Either isoform of the dopamine D2 receptor can mediate dopaminergic repression of the rat prolactin promoter

Ruth McChesney, Stuart C. Sealfon, Manami Tsutsumi, Ke Wen Dong, James L. Roberts, Carter Bancroft

Research output: Contribution to journalArticlepeer-review

24 Scopus citations


Hypophyseal portal dopamine is a major negative regulator of pituitary prolactin (PRL) production. Dopamine has been reported to repress PRL gene transcription in pituitary cells. To facilitate further study of the effect of dopamine on PRL gene activation, we introduced PRL promoter and D2 receptor (D2R) constructs into GH3 cells. Since two D2R isoforms (termed D2S and D2L) have been cloned previously, we first determined which isoform(s) is present in the lactotroph by measuring the level of each mRNA species in rat prolactinoma. mRNA for each D2R isoform was found to be present, with the D2L mRNA in great (c. 6-fold) excess. Because the lactotroph contains both isoforms, the effect of each on the PRL promoter was investigated. The cDNA for each receptor isoform was synthesized by polymerase chain reaction, and cloned into an RSV-based expression vector. GH3 cells were then transiently co-transfected with either of the resulting RSV-D2R constructs plus a PRL-chloramphenicol acetyltransferase (CAT) construct containing the first 1957 base-pairs of PRL gene 5'-flanking DNA. The cells were then incubated 48 h plus or minus the dopamine agonist ergocryptine (ECR). In the presence of either RSV-D2R isoform, ECR yielded a 4-5-fold decrease in CAT activity, an effect not seen in the absence of the RSV-D2R. The promoter specificity of this effect was demonstrated by the inability of ECR to regulate expression of a control RSV-CAT construct. The PRL promoter repression mediated by each receptor isoform had appropriate pharmacology: the specific D2R agonist, quinpirole, yielded results similar to ECR, and the ECR repression was reversed by the dopamine antagonist spiperone. These data imply that rat PRL 5'-flanking DNA contains a dopamine response element(s) which can respond to dopaminergic signals mediated by either D2R isoform.

Original languageEnglish (US)
Pages (from-to)R1-R7
JournalMolecular and Cellular Endocrinology
Issue number1-3
StatePublished - Aug 1991
Externally publishedYes


  • Dopamine
  • Prolactin gene
  • Transfection

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Endocrinology


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