Several studies have demonstrated that cyamemazine, a classic antipsychotic compound, possesses anxiolytic properties in humans as well as a potent antagonistic effect on 5-HT(2C) and 5-HT(3) receptors. In this study the level of anxiety of mice was assessed in the light/dark exploration test and the elevated plus maze (EPM) following both acute and chronic administration. Spontaneous locomotor activity was measured using a photoelectric actimeter. Acute or chronic administration of cyamemazine dramatically decreases the spontaneous locomotor activity of mice at the dose of 1 mg/kg in comparison with the control group. In the light/dark exploration test, cyamemazine (0.375 mg/kg) only demonstrated anxiolytic-like activity following acute administration. In the elevated plus maze (EPM), cyamemazine did not induce any anxiolytic like effects after acute administration. However, after chronic administration, cyamemazine at doses of 0.25, 0.375, 0.5 and 1 mg/kg significantly increased the time spent in the open arms. The number of open arm entries was also increased at 0.25 and 0.5 mg/kg. Various serotonergic ligands were then used to examine the role of the various receptors in mediating the effects of cyamemazine in the EPM. Concerning the 5-HT(2) ligands DOI and mCPP antagonised the effects of cyamemazine and N-desmethyl clozapine potentiated the effects. For 2-methyl-5-HT and ondansetron, the 5-HT(3) receptor ligands did not seem to have any effect. It appears that the 5-HT(2C) receptors are more implicated in the function of mediating the anxiolytic effect of cyamemazine in the EPM. The discrepancy of results obtained in the tests, following acute or chronic administration could be the result of the combined activity of dopamine D(2) receptor antagonism with antagonism of 5-HT(2C) and 5-HT(3) receptors.