Experimental design of behavioral studies in animals generally includes placebo-treated controls. However, when placebo is administered by injection in experimental models of psychiatric diseases such as depression, where stress may affect the execution of the behavioral test, it is possible that injection per se may influence the behavioral response. Rats injected with clomipramine hydrochloride (1, 10 or 50 mg/kg), as compared to animals injected with physiological saline as placebo, showed a dose-dependent decrease of the immobility time in the despair test and of the number of floor units explored in the open field in the reserpine test. However, when animals injected with placebo or clomipramine 50 mg/kg were compared with untreated intact controls, it was found that the immobility time in the despair test was higher in the placebo-treated animals than in untreated intact controls. A difference was found between clomipramine-injected animals and untreated intact controls. In contrast, rats tested in the reserpine test, which is based on repeated drug injections, no difference was found between placebo-treated animals and untreated intact controls. These results indicate that stressful procedure of the experimental design may change the response of animals in behavioral tests. Studies with experimental models of depression, where stressful procedures are used, should include a control group of untreated intact animals.