Abstract: Acute i, p injection of diazepam (3 mg/kg) significantly decreased the open field behaviors (locomotion and rearing) of rats. However, when diazepam (3 mg/kg) was given ip daily for 15 consecutive days, no depressive effect on the open field behaviors was observed. After the open field observation, rats were immediately decapitated and brain slices of cortex, hippocampus, amygdala and brain stem were stimulated by high K⁺ (65 mmol/L) to cause the release of noradrenaline (NA), dopamine (DA) and serotonin (5-HT). Diazepam (9 μmol/Ls) ignificantly depressed the release of monoamine neurotransmitters caused by high K⁺, which were antagonized by Ro 15-1788 (5 μmol/L) in vitro. Acute administration of diazepam (ip 3 mg/kg) also significantly depressed the release of monoamine neurotransmitters caused by high K⁺ from all regions observed. However, hippocampus showed tolerance to the depressive effects of diazepam on the releases of NA and DA, and amygdala only on the release of DA, caused by high K⁺, when given ip chronically (3 mg/kg, 15 d). Our results suggest that the depressive effect of benzodiazepines on the release caused by anxiogenic factors of catecholamine in the hippocampus, front cortex and amygdala regions may be one of the mechanisms of sedative and anxiolytic actions of benzodiazepines.