The brain serotonin is involved in mediation of emotional behaviour including anxiety and related fear conditioning. It is known that the median raphe nucleus (MRN) is the origin of a serotonergic pathway and mainly innervates septo-hippocampal formation which plays an important role in emotional cognition. However, its regulatory role in different types of fear conditioning is still unclear. In the present study, the animals underwent ibotenic acid or sham lesions of the median raphe nucleus and the effects of MRN lesions on immediate and delayed fear conditioning to multiple contextual cues were studied. Freezing behaviour served as a measure of contextual fear. Sham-lesioned animals showed reliable conditional freezing when observed immediately following foot-shock (1.0 mA) for 3-min test and 48 h after the shock for 12-min test. Rats with MRN lesions displayed robust freezing behaviour immediately after the shock, even though they showed a marked deficit in freezing 48 h following the shock. These findings indicate that the MRN-serotonergic septo-hippocampal pathway is involved in the regulation of anxiety related to fear conditioning triggered by contextual cues, suggesting that short-term contextual fear is independent on the MRN while long-term contextual fear depends on the MRN. (C) 2000 Elsevier Science B.V. All rights reserved.