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The major goal of the present study was to examine acute tolerance to nicotine-induced disruption of operant behavior following a single, noncontingent injection. Rats were trained to lever press for food reinforcement under a fixed ratio-30 schedule. Once trained, rats were injected with either saline or nicotine (0.8 mg/kg) in their home cages. After either a 90- or 180-min delay, each rat was injected with nicotine (0.4 mg/kg) and placed in the operant chamber for a 30-min behavioral evaluation session. This experiment was replicated with slight modifications 1 week later. The results of the present study suggest that 0.8 mg/kg of nicotine produces acute tolerance to the response rate decreasing effects of 0.4 mg/kg of nicotine. Because the tolerance-producing dose of nicotine was injected while rats were not in the test environment, they did not have an opportunity to practice the target behavior while under the influence of the drug. Hence, the acute tolerance observed in this study appears to be, at least partly, pharmacological (vs. behavioral) in nature, and may be related to a desensitization of central nicotinic acetylcholinergic receptors (nAChRs)