1. An extracellular recording technique was used to study the effects of 5-hydroxytryptamine (5-HT, serotonin) on the tetanus-induced long-term potentiation (LTP) of the nicotinic pathway of transmission in the superior cervical ganglion (SCG) of the rat. The postganglionic compound action potential (CAP), made submaximal by treatment with hexamethonium (O.4 mM), was used as an index of transmission in the ganglion. 2. Serotonin (10 microM) markedly enhanced the magnitude of LTP without affecting the post-tetanic potentiation (PTP). The serotonin (2-30 microM) concentration-response curve for LTP was bell shaped as no enhancement was seen with 30 microM serotonin. This may largely be due to activation of a 5-HT1 receptor subtype and not to desensitization. 3. When superfused before tetanus, the 5-HT1A receptor agonist 8-hydroxydipropylamino-tetralin (8-OH-DPAT, 5 microM) prevented the expression of LTP without affecting PTP. 4. Pretreatment of ganglia with the 5-HT2 receptor agonist R-(+)-dimethoxy-4-iodoamphetamine (R-(+)-DOI, 1 microM) enhanced the tetanus-induced LTP. Similar treatment with the 5-HT2 receptor antagonist ketanserin (3 microM) had no significant effect on LTP. 5. Pretreatment of ganglia with the 5-HT3 receptor agonist 1-m-(chlorophenyl) biguanide (m-CPGB, 1 microM), markedly increased (300%) the tetanus-induced LTP. Similar pretreatment with the 5-HT3 receptor antagonist 3-tropanyl-3,5-dichlorobenzoate (MDL 72222, 0.5 microM) completely prevented the expression of LTP. Fully expressed LTP was reversibly blocked by MDL 72222 when applied during the maintenance phase of LTP. 6. Tetanic stimulation of monoamine-depleted ganglia (from reserpine-pretreated rats, 3 mg kg-1 for 24 h) failed to induced LTP. 7. In monoamine-depleted ganglia, tetanus preceded by superfusion with m-CPBG readily induced LTP. MDL 72222 completely blocked this LTP. However in these ganglia tetanus failed to induced LTP when m-CPBG was given 2 min (during PTP) or 1 h after tetanus. 8. Tetanic stimulation of monoamine-depleted ganglia in the presence of R-(+)-DOI failed to induced LTP. 9. We conclude that tetanus-induced LTP of the SCG of the rat requires activation of 5-HT3 receptors both for induction and maintenance.