1. The aim of the present study was to investigate the origin and modulation of 5-hydroxytryptamine release in dorsal hippocampus of freely moving rats using brain microdialysis. 2. Basal release of 5-hydroxytryptamine and 5-hydroxyindole-acetic acid was 3.0 +/- 0.2 pg/15 min and 519 +/- 18 pg/15 min, respectively. Stimulation by 60 mM K+ increased 5-hydroxytryptamine release by 66%. Inclusion of 10 microM tetrodotoxin in the perfusion medium reduced 5-hydroxytryptamine release to approximately 5% of basal levels. 3. In addition, release could be modulated by the 5-hydroxytryptamine receptor agonist 8-hydroxy-2-(di-N-propylamino)-tetralin (50 % decrease) and this effect was completely blocked by the 5-hydroxytryptamine-1 receptor antagonist pindolol. 4. These data are in concordance with results from the ventral hippocampus and indicate that release of 5-hydroxytryptamine is predominantly of neuronal origin. In contrast with other studies, the authors found the non-selective 5-hydroxytryptamine-1 antagonist, pindolol, to increase 5-hydroxytryptamine release by 65% following systemic administration. This may hint at a different autoreceptor control in serotonergic fibers ascending from the median raphe as compared to the dorsal raphe nucleus.