The prelimbic cortex arbitrates memory-guided conflict resolution.

Balancing reward and avoidance to produce adaptive behavior is a critical aspect of conflict resolution. Here, we investigated how mice balance reward-seeking and avoidance during memory-guided conflict resolution. Mice were trained to integrate the drives to approach a reward and to avoid a foot shock without compromising either goal. During training, mice learn that a light cue signals the availability of a reward and that an auditory cue signals the onset of a foot shock, which they can avoid by stepping onto a platform. During testing, the light and auditory cues are delivered simultaneously, creating a conflict between reward-seeking and punishment avoidance. Our results demonstrate that mice learn to flexibly accommodate behavioral drives to maximize reward while avoiding the foot shock. We then leveraged a whole-brain tissue clearing approach to identify whole-brain activity profiles and specifically identified the prelimbic cortex (PL) as a central hub. Using bulk calcium imaging, we found PL activity signals task variables and predicts successful conflict resolution. Silencing PL engrams disrupts the balance between reward-seeking and avoidance behaviors. By leveraging single-cell calcium imaging, we reveal that the PL integrates valence-specific representations via distinct functional network patterns, and disruptions in network modularity and network roles contribute to reduced behavioral flexibility during conflict. Finally, silencing ventral tegmental area (VTA)-PL-projecting neurons uncovered a new role in VTA neurons in supporting PL functionality during memory-guided conflict resolution. Together, these findings reveal the PL's mechanistic role in guiding behavioral flexibility between memory-guided behaviors during conflict.