Leptin and neuropeptide-Y (NPY) are indicated to play a role in hypothalamo-pituitary-thyroid (HPT) axis in relation to regulation of energy homeostasis mediated through acting at hypothalamic synthesis and release of thyrotropin (TSH)-releasing hormone (TRH). Whether leptin and NPY also act at pituitary level in HPT axis remains unknown. This study aimed at investigating whether or not leptin and NPY exert actions at pituitary in modulation of transcript levels of TSHbeta and the common pituitary glycoprotein hormone alpha (PGHalpha) subunits. The dispersed pituitary cells from 6 wk old male Wistar rats were incubated with or without TRH, leptin or NPY of 10(-8) M and 10(-10) M for 6 h at 37 degrees C in medium-199 under aeration of 95% O2 and 5% CO2. The mRNA levels of TSHbeta and PGHalpha subunits of the incubated cells were measured by reverse transcription-polymerase chain reaction. The results revealed that leptin stimulated, while NPY inhibited, TSHbeta mRNA levels in a dose-related manner. Both leptin and NPY increased alpha subunit mRNA levels. It is demonstrated for the first time that both leptin and NPY exert a direct action at rat pituitary affecting steady-state levels of mRNA of TSHbeta and PGHalpha subunits. The present study supports that both leptin and NPY act at the pituitary as well besides the hypothalamus in HPT axis in relation to regulation of energy homeostasis.