A pituitary-specific trans-acting factor, Pit-1 regulates transcriptional activity of growth hormone (GH) and prolactin (PRL) genes. Pit-1 can bind and activate the promoters of human chorionic somatomammotropin (hCS-A) and placental GH variants (hGH-V) as well. However, expression of Pit-1 in the rat placenta has not yet been elucidated. The present study aims to determine whether the Pit-1 gene is locally expressed in the rat placenta using reverse transcription-polymerase chain reaction (RT-PCR), Northern blot and Western blot hybridization, in situ hybridization and immunohistochemistry. PCR products were further analyzed by Southern hybridization and DNA sequencing. The estimated size of Pit-1 mRNA in placenta was very similar to that in anterior pituitary (AP). PCR products from placenta were exactly the same size with that from AP and confirmed as Pit-1-specific by Southern hybridization. The Pit-1 specific sequence was also confirmed by sequencing of partial amplification fragments. Immunoreactive 33 kDa Pit-1 was present in the placenta as well as in AP. Pit-1 specific mRNA and protein were localized in the trophoblast cells of placenta. These data suggest that Pit-1 is locally synthesized in the rat placenta and may be involved in the regulation of GH- and/or PRL-like gene expression in the placenta.