Phasic dopamine (DA) release is believed to guide associative learning. Most studies have focused on projections from the ventral tegmental area (VTA) to the striatum, and the action of DA in other VTA target regions remains unclear. Using optogenetic activation of VTA projections, we examined DA function in the medial prefrontal cortex (mPFC). We found that mice perceived optogenetically induced DA release in mPFC as neither rewarding nor aversive, and did not change their previously learned behavior in response to DA transients. However, repetitive temporal pairing of an auditory conditioned stimulus (CS) with mPFC DA release resulted in faster learning of a subsequent task involving discrimination of the same CS against unpaired stimuli. Similar results were obtained using both appetitive and aversive unconditioned stimuli, supporting the notion that DA transients in mPFC do not represent valence. Using extracellular recordings, we found that CS-DA pairings increased firing of mPFC neurons in response to CSs, and administration of D 1 or D 2 DAreceptor antagonists in mPFC during learning impaired stimulus discrimination. We conclude that DA transients tune mPFC neurons for the recognition of behaviorally relevant events during learning.learning | attention | dopamine T he firing activity of dopamine (DA) neurons in the ventral tegmental area (VTA) is consistent with a role of reward prediction error signal, which is believed to guide behavioral adaptation through DA release in target brain regions (1-7). Experiments using optogenetic manipulations have established a causal link between the activity of DA neurons in VTA and the reinforcing signal that mediates learning and conditioning (8-10). Stimulation of VTA, however, results in transient DA release in many target areas, and it is unclear how each of these regions contributes to learning. For example, compared with the striatum, the medial prefrontal cortex (mPFC) receives fewer DA projections (11), expresses fewer DA reuptake transporters (12), and exhibits an overall lower level of DA (13,14). Nevertheless, pharmacological studies have implicated DA as a powerful neuromodulator of mPFC, able to influence many cognitive functions that support learning (15-21). Furthermore, DA neurons in VTA projecting to either the striatum or mPFC have distinct intrinsic neuronal properties and receive distinct inputs (22-24); thus, they are likely to serve different roles. It remains unclear what function phasic DA release might have in mPFC, whether it carries any valence, or how it affects stimulus-specific learning.In this study, we addressed specifically the role of phasic DA release in stimulus-response association. Using optogenetically timed release of DA from VTA neuronal projections in mPFC, we found that DA transients enhance the firing of mPFC neurons in response to the paired conditioned stimuli (CSs), whereas blocking DA receptors in mPFC during learning impairs stimulus discrimination. Furthermore, pairing a CS with optogenetic stimulation of DA fibers in ...