TRH gene expression in hypophysiotropic neurons of the hypothalamic paraventricular nucleus (PVN) is under regulation by thyroid hormone circulating in the bloodstream. To determine whether thyroid hormone could exert effects directly on TRH-producing neurons in the PVN, the presence of thyroid hormone receptors (TR) in these neurons was determined by double labeling immunocytochemical techniques, using specific antiserum to each of the functional TRs, TR alpha 1, TR beta 1, and TR beta 2, followed by antiserum to prepro-TRH-(25-50) as a marker for TRH neurons. In addition, the presence of the TR variant, TR alpha 2, was sought in these cells. Immunoreactive TR alpha 1 and TR beta 2 were found in the greatest percentage of TRH neurons in the PVN (91.1 +/- 2.5% and 83.8 +/- 2.1%) and intensely stained the nucleus. Immunoreactive TR beta 1 was also found in the majority of TRH neurons, but stained PVN cells only lightly compared to the other TRs. TR alpha 2 was found to coexist in only a minority of TRH neurons in the PVN and also lightly immunostained the nucleus compared to its more intense labeling in other regions of the brain. We conclude that hypophysiotropic TRH neurons contain functional TRs, and therefore, these neurons could be directly influenced by thyroid hormone. The relative paucity of TR alpha 2 in these cells could contribute to the selectivity of this population of TRH neurons to the effects of circulating levels of thyroid hormone.