Feeding a novel food (Fonzies) to rats fed ad libitum with standard food increased extracellular dopamine (DA) in the medial prefrontal cortex (PFCX) and in the medial nucleus accumbens (NAc). Previous Fonzies feeding, although it did not affect the increase of extracellular DA in the PFCX in response to Fonzies feeding, blunted that increase in the NAc (habituation); recovery from habituation in the NAc was complete 5 d after previous Fonzies feeding. Predictive association of an otherwise neutral stimulus extrinsic to Fonzies (empty plastic box) with Fonzies feeding resulted in the acquisition by the stimulus of the property to elicit incentive responses directed toward the stimulus and to increase extracellular DA in the PFCX. However, the same stimulus, or a more complex stimulus including intrinsic stimuli (Fonzies-filled plastic box), failed to acquire the ability to modify extracellular DA in the NAc. Pseudoconditioning, i.e., nonpredictive association of the extrinsic stimulus (empty box) with Fonzies feeding, did not result in acquisition by the stimulus of the property to elicit incentive responses and to increase extracellular DA in the PFCX. Repeated nonreinforced presentation of previously conditioned extrinsic stimuli (empty box) resulted in extinction of the property to elicit incentive responses and to increase extracellular DA in the PFCX. These results indicate that in rats fed ad libitum, phasic activation of mesocortical and mesolimbic DA systems by motivational stimuli is differentially influenced by associative (conditioning) and nonassociative (habituation) learning mechanisms and is differentially related to acquisition and expression of incentive motivation.