Immunohistochemical evidence indicates the coexistence of serotonin and TRH in many raphe neurons. We examined the biochemical changes in TRH receptors after destruction of the serotonergic pathways by 5,7-dihydroxytryptamine (5,7-DHT). 2 weeks after an intracerebroventricular injection of 5,7-DHT, rats were killed, and specific brain regions were dissected on ice. Serotonin levels in the CNS of lesioned rats was reduced by 50-85% in all regions, with the highest reduction in the spinal cord and hippocampus. Immunoreactive TRH was reduced in the spinal cord by 70%, but other brain regions contained normal levels of TRH. TRH receptor binding was increased by 40% in the spinal cord of lesioned rats, but appeared unchanged in rostral brain regions in which no decrease in TRH content was detected. Scatchard plots of TRH receptor binding in the spinal cord indicated that the increased binding after 5,7-DHT administration reflected an increased receptor number. These findings suggest that biochemical up-regulation of TRH receptors occurs in the spinal cord following depletion of TRH.