The effects of acute physiological stressors (5 s tail pinch, handling and forced swimming at +25 and +5 degrees C for 3 min each) on serotonin (5-HT) release in the mouse brain were investigated using in vivo microdialysis. The extracellular 5-HT levels were determined by a newly developed highly-sensitive and selective high-performance liquid chromatography method based on derivatization with benzylamine and fluorescence detection. The basal levels of 5-HT in 3 min microdialysates from the ventral hippocampus and frontal cortex were 0.68+/-0.21 and 0.75+/-0.28 fmol/6 microl (n=24), respectively. All three stressors caused an immediate, significant and reversible increase (handling: 150%; swimming: 240%) of extracellular 5-HT levels in both brain structures, suggesting a more dynamic role played by the serotonergic system in response to acute stress.