Systemic administration in rats of p-chloroamphetamine (PCA; 2 x 10 mg/kg) reduced the in vitro uptake of 14C-5-hydroxytryptamine (14C-5-HT) in cortical synaptosomes by 76% and in spinal cord synaptosomes by 35%. Intrathecal injection of 5,6-dihydroxytryptamine (20 micrograms/rat) selectively lesioned the descending serotonergic pathways (83% reduction in uptake of 14C-5-HT in spinal synaptosomes, no significant change in uptake in cortical synaptosomes). Administration of PCA or 5,6-DHT did not significantly alter the uptake of 3H-noradrenalin into cortical or spinal synaptosomes. The response thresholds of the rats in the increasing temperature hot plate test (1 to 7 days after administration) were unaffected by either type of lesion. Interference with the antinociceptive effect of PCA (2.5 mg/kg) was evaluated 7 days after administration of the neurotoxins. PCA pretreatment strongly reduced the peak of the PCA-induced antinociception while 5,6-DHT reduced its duration. Thus, both ascending and descending serotonergic pathways contribute to PCA-induced antinociception.