The 5-HT1B subtype of serotonin (5-HT) receptor regulates the release of 5-HT in the rat spinal cord. In an attempt to confirm the presynaptic location of the 5-HT1B receptor, serotonergic nerves were destroyed neurochemically with 5,7-dihydroxytryptamine (5,7-DHT) administered intrathecally. At 3, 7 and 14 days post 5,7-DHT, competitive radioligand binding assays were performed using 2 nM [3H]5-HT and varying concentrations of trifluoromethylphenylpiperazine (TFMPP), a drug which interacts with 5-HT1B and 5-HT1A binding sites (affinity of 5-HT1B site greater than 5-HT1A site for TFMPP). The decrease in 5-HT1B sites that was expected was not observed at any time following the lesion. However, the binding capacity of the 5-HT1B site, as well as the 5-HT1A site, increased significantly by 7 and 14 days postlesion, respectively. Thus, it appears that an up-regulation of 5-HT1B sites occurred that may have obscured the ability to observe a reduction in a small population of sites located on the serotonergic nerve terminals.