Autoregulation of the rat prolactin gene in lactotrophs

The autoregulation of prolactin (PRL) secretion in the rat has been demonstrated at both the hypothalamus and the pituitary levels. Studies on the direct negative feedback effect of PRL in the lactotrophs have concentrated on the acute effect on PRL secretion which does not involve change in PRL synthesis. In this study, we have developed a cotransfection assay in somatolactotrophs where we examine the effect of PRL on the transcription of its own gene. We found that oPRL, at physiological concentrations, exerts a strong and specific inhibition of the rPRL gene transcription in PRL-deficient GC cells. This effect is mediated by both the intermediate and the long forms of PRL receptor. The inhibition was also reproduced in GH3 cells, which secretes PRL, by adding exogenous oPRL in the presence of anti-rat PRL antiserum to neutralize endogenous rPRL. Cellular specificity was demonstrated by testing this regulation in non-pituitary cell types where no modulation of the PRL promoter reporter gene could be elicited by PRL, even with cotransfection with the Pit-1 expression vector. Finally, deletions of the rPRL promoter indicate that the full inhibitory effect of PRL requires the same regulatory domains (proximal and distal) that have been described for the other PRL gene regulators. These results strongly suggest the existence of the extra-short loop regulation of the rat PRL at the transcriptional level.