Traditional operant self-administration and conditioned place preference methods have yielded inconsistent results in studies of nicotine reinforcement thereby hindering efforts to identify the neurobiological systems underlying the drug's motivation and reinforcement. This study was designed to assess the motivation of subjects to seek nicotine using a runway self-administration procedure. Male Sprague-Dawley rats (N=67) were trained to run a straight alley for a single daily intravenous injection of nicotine (0.01-0.09 mg/kg/injection) on each of 21 consecutive trials. Run Speed (1/Run Time) served as the dependent measure for the animals' motivation to traverse the alley and enter a goal-box associated with intravenous nicotine administration. Nicotine induced an inverted U-shaped dose-response curve with the 0.03 mg/kg dose producing optimal runway performance over trials. Subjects running for doses larger or smaller than the optimal dose exhibited slower running and took longer to enter the goal-box. Thus, the runway procedure proved to be an effective methodology for reliably assessing the motivation of trained but nondrugged animals to seek intravenous nicotine.