Pituitary-specific transcription factor Pit-1 regulates growth hormone and prolactin gene expression in the pituitary. Recently, Pit-1 was shown to be locally synthesized in the rat placenta and is involved in the regulation of rat placental lactogen (PL) gene expression. Pit-1 has three different splicing variants. They are well known as being biologically active. In the present study, we found that Pit-1 beta is also synthesized in the rat placenta and we tried to examine the effects of dopamine and estrogen on the regulation of Pit-1 alpha, beta and PL-II genes expression using the reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot hybridization. A dopamine receptor agonist, bromocriptine, significantly decreased placental Pit-1 alpha, beta, and PL-II mRNA levels. To examine the effect of estrogen on the gene expression of Pit-1 beta, pregnant female rats were ovariectomized (OVX) and injected daily with 17 beta-estradiol. OVX markedly lowered the amount of Pit-1 beta mRNA. Estrogen injection recovered the OVX-induced inhibition of Pit-1 beta mRNA level. Finally, we investigated the site-specific transcription of Pit-1 alpha and beta mRNA in the labyrinth zone and junctional zone of the placental in 15 and 20 days of gestation. The main site of Pit-1 alpha and beta synthesis was shifted from the junctional zone to the labyrinth zone from 15 to 20 days of gestation. Together, these data presume that Pit-1 beta may play a more important role in the placenta than in the pituitary and that Pit-1 may be involved in the regulation of the PL and prolactin-like peptide by estrogen and dopamine in the rat placenta.