S U M M A R YThe pars tuberalis (PT) of the pituitary represents an important target site for the time-pacing pineal hormone melatonin because it expresses a large number of mt1 receptors. Functional studies suggest that the PT mediates the seasonal effects of melatonin on prolactin (PRL) secretion. The aim of this study was the characterization of the phenotype of melatonin-responsive cells. Furthermore, we determined whether ROR  , a retinoid orphan receptor present in the PT, was co-expressed in the same cells. We combined nonradioactive in situ hybridization (ISH) with hapten-labeled riboprobes for detection of the receptors and immunocytochemistry (ICC) for detection of ␣ GSU ( ␣ -glycoprotein subunit),  TSH,  FSH,  LH, GH, PRL, and ACTH. Expression of mt1 mRNA was found in small round cells, co-localized with ␣ GSU and  TSH. However, not all  TSH-containing cells expressed mt1 mRNA. The distribution of mt1-and ROR  -positive cells appeared to overlap, although more cells were labeled for ROR  than for mt1. Gonadotrophs, as well as other pars distalis cell types, were never labeled for mt1 melatonin receptor. Therefore, this study identifies the "specific" cells of the PT as the mt1 melatonin receptor-expressing cells.(