The microdialysis technique was used to study the ability of essential oil from citrus lemon to modulate hippocampal acetylcholine (ACh) release in male and female rats. Animals were allowed to inhale this odor while experiencing a persistent nociceptive input (50 microl formalin, 5%) or under control conditions (sham-injection). In males, exposure to the essential oil did not change the time course and magnitude of the ACh increase induced by pain. In females, the pain-induced increase of ACh was delayed and increased by exposure to lemon essential oil. The present results indicate that lemon essential oil affects the ACh release differently in male and female rats during a painful condition.