To examine the role of the nitric oxide (NO) pathway in the stimulus effects induced by some psychostimulants, separate groups of rats were trained to discriminate between amphetamine (AMPH; 0.5 mg/kg) and saline, or cocaine (COC; 5 mg/kg) and saline using a standard two-lever operant procedure. Substitution studies showed that AMPH and COC generalized for the training drugs in a dose-dependent manner, their ED50, values being 0.1 mg/kg and 1.2 mg/kg, respectively. The dose-response function of both those psychostimulants did not change in the course of the experiment. Moreover, AMPH and COC induced cross-substitution effects towards each other. Successive combination tests demonstrated that injection of a fixed dose of the NO synthase (NOS) inhibitor 7-nitro indazole (7-NI; 25 mg/kg) plus different doses of AMPH or COC resulted in a leftward shift in the dose-response curves of those psychostimulants and a decrease in their ED50 values. On the other hand, pretreatment with the NO donor molsidomine (MOL), injected in a fixed dose of 100 mg/kg before AMPH and COC, shifted the dose-response curves of the psychostimulants to the right and increased their ED50 values. Our results indicate that NO plays an inhibitory role in the dopamine (DA)-evoked discrimination effects of AMPH and COC in rats.