Serotonin (5-hydroxytryptamine; 5-HT) plays a role in the modulation and processing of pain and evidence has been provided that some nonsteroidal anti-inflammatory drugs (NSAIDs) act, at least in part, through this system. The present study was designed to investigate the possible participation of 5-HT 1 , 5-HT 2 , 5-HT 3 , and 5-HT 7 receptor subtypes in the antinociceptive effect of S(þ)-ketoprofen (S-KP) at spinal and supraspinal level using the "pain induced functional impairment model in rat" (PIFIR model). S-KP was administered orally (p.o.) and antagonist drugs for 5-HT receptors (5-HT 1 /5-HT 2 , and 5-HT 3 ) were administered intrathecally (i.t.) or intracerebroventricularly (i.c.v.) 15 min before S-KP. S-KP (3.4 mg/kg p.o.) produced a significant antinociceptive effect in this model. Pre-treatment with the 5-HT 1 / 5-HT 2 /5-HT 7 receptor antagonist, methiothepin (1.5 mg, i.c.v.), significantly reversed the antinociceptive effect of S-KP. In contrast, no significant differences were observed following i.t. administration of methiothepin. Pre-treatment i.t., but not i.c.v., with the 5-HT 3 /5-HT 4 receptor antagonist, tropisetron (0.9 mg), on the other hand, significantly reversed S-KP-induced antinociception. These results indicate that serotonin mechanisms are involved in the antinociceptive effect of S-KP. 5-HT 1 /5-HT 2 /5-HT 7 receptors participate at the supraspinal level and 5-HT 3 /5-HT 4 receptors participate at spinal level. Drug Dev.