Frontal noradrenergic and cholinergic transients exhibit distinct spatiotemporal dynamics during competitive...

Norepinephrine (NE) and acetylcholine (ACh) are crucial for learning and decision-making. In the cortex, NE and ACh are released transiently at specific sites along neuromodulatory axons, but how the spatiotemporal patterns of NE and ACh signaling link to behavioral events is unknown. Here, we use two-photon microscopy to visualize neuromodulatory signals in the premotor cortex (medial M2) as mice engage in a competitive matching pennies game. Spatially, NE signals are more segregated with choice and outcome encoded at distinct locations, whereas ACh signals can multiplex and reflect different behavioral correlates at the same site. Temporally, task-driven NE transients were more synchronized and peaked earlier than ACh transients. To test functional relevance, we stimulated neuromodulatory signals using optogenetics to find that NE, but not ACh, increases the animals' propensity to explore alternate options. Together, the results reveal distinct subcellular spatiotemporal patterns of ACh and NE transients during decision-making in mice.