Reward uncertainty attributes incentive value to reward proximal cues, while amphetamine sensitization reverts attention to more predictive reward distal cues.

Slot-machine gambling incorporates numerous audiovisual cues prior to and during reward delivery (e.g. spinning wheels, flashing lights, celebratory sounds). Over time, these cues may motivate playing and even elicit cravings and relapse in those affected by gambling disorder. Animal studies suggest a heightened attraction to these cues despite diminished predictive ability under reward uncertainty, as evidenced by sign-tracking behavior in rats. Repeated amphetamine administration may also enhance the incentive value attributed to cues. Here, we explored the impact of reward uncertainty and prior amphetamine sensitization on the relative attractiveness and conditioned reinforcing properties of serial Pavlovian cues with different degrees of predictive and incentive value in rats. Animals were sensitized through repeated injections of amphetamine (1-4 mg/kg) or saline and then trained in a Pavlovian autoshaping task involving two sequential lever-auditory cue combinations (CS1, CS2) under Certain (100%-1) or Uncertain (50%-1-2-3) reward conditions. Subsequently, we evaluated the impact of acute amphetamine exposure on cue attraction. Our results suggest that Uncertainty alone enhanced attraction towards the reward-proximal CS2. However, combined sensitization and Uncertainty reversed cue preference relative to Uncertainty alone, enhancing attraction towards the more predictive reward-distal CS1. Both cues acquired conditioned reinforcing properties, despite the CS2 being otherwise ignored in all groups besides Uncertainty. However, combined sensitization and Uncertainty increased the reinforcing value of both cues and doubled the amount of interaction with the CS1 lever per presentation. Our results imply competitive mechanisms for attributing incentive value to gambling-related cues between reward uncertainty, prior amphetamine sensitization, and acute amphetamine administration.