Diazinon, an organophosphate pesticide, is relatively highly toxic to fish and causes vertebral malformation and behavioral change of fish at relatively low concentrations. To elucidate biochemical mechanism of the behavioral change of Oryzias latipes (killifish) caused by diazinon, the effect of the insecticide on acetylcholine esterase activities and the levels of some neurotransmitters were evaluated. Acetylcholine esterase activities in both head and body were significantly lowered at the concentration of 10 ppb of diazinon and acetylcholine contents in head tended to be upregulated with increasing concentration of diazinon. Exposure of killifish to 5000 ppb diazinon resulted in gradual decrease in acetylcholine content in body part with exposure time. Norepinephrine and serotonin concentrations in killifish head and body were highest at 1000 ppb of diazinon while neurotransmitter were relatively low in fish unexposed or exposed to lower dose of the pesticide, suggesting that increased norepinephrine and serotonin can partially account for diazinon-induced behavioral abnormality.