Pharmacological studies suggest that the discriminative stimulus (DS) produced by nicotine is mediated centrally. The aim of the present study was to identify neuroanatomical substrates that mediate the DS properties of nicotine. Specifically, the nucleus accumbens, a brain region known to mediate the DS effects of amphetamine and cocaine, was investigated using a two-lever operant drug discrimination paradigm. Male hooded rats were trained to discriminate nicotine (0.2 mg/kg s.c.) from saline with a tandem schedule of food reinforcement. Once stimulus control was attained, a randomised sequence of nicotine microinjections (2-8 micrograms) was tested for generalisation during brief extinction tests. It was confirmed that the stimulus produced by the systemic administration of nicotine generalized to nicotine administered bilaterally into the dorsal hippocampus, with significant decreases in overall response rates. Microinjections of nicotine (1-8 micrograms) into the nucleus accumbens failed to produce any dose-related increases in responding on the nicotine-appropriate lever although these microinjections also produced significant decreases in response rates. Smaller doses (1-4 micrograms) of nicotine administered into the fourth ventricle produced characteristic prostration responses but these microinjections failed to produce generalization in tests carried out 20 min later, when the disabling effects of prostration had dissipated. These results suggest that the DS effects of nicotine may be mediated, at least in part, through the dorsal hippocampus. Results from intra-accumbens and intraventricular injections suggest that these regions may not be important in mediating the DS effects of nicotine.