Neurons containing thyrotropin-releasing hormone (TRH) and glucocorticoid receptor (GR) immunoreactivity (IR) were demonstrated by a two-colour immunoperoxidase method in coronal cryotome sections of the preoptic region and the hypothalamus of the male rat brain. All the TRH-IR neurons (TRH-IR) located in the dorsal hypothalamus - medial and dorsal parvocellular parts of the paraventricular hypothalamic nucleus and the dorsomedial hypothalamic nucleus - and in the anterior periventricular hypothalamic nucleus were strongly GR-IR. The TRH-IR neurons of the medial preoptic area, the perifornical nucleus and the medial tuberal area were mostly weakly GR-IR and some lacked GR-IR. These data indicate a differential regulation of diencephalic TRH-IR neurons by glucocorticoids. They also imply that the inhibitory effect of glucocorticoids on TSH secretion may involve a direct inhibition of TRH synthesis and/or release by a nuclear action in the TRH-IR nerve cells of the paraventricular hypothalamic nucleus projecting to the median eminence.