Title: Conjoint outcome prediction and reporting by ACh, DA, NE, & 5-HT in primary visual and auditory cortices

This study explores the intricate interplay between acetylcholine and calcium signaling within the cortical mantle, specifically focusing on primary sensory cortices. Despite their recognized impact on cortical plasticity, the coordination of their release and their collective influence on learning and memory processes remain poorly understood. Simultaneous measurements of neuronal activities and acetylcholine transients, particularly in primary sensory cortices, allow us to quantify the relationship between behavioral state, cortical activity, and neuromodulatory signaling, and evaluate their spatiotemporal release profiles. Using a dual-color wide-field imaging setup, we monitored the activity of acetylcholine and calcium across the dorsal cortex in head-fixed awake mice engaged in appetitive conditioning tasks. The red-fluorescent calcium indicator jRCaMP1b and the green-fluorescent ACh indicator ACh3.0 were employed to assess neural responses during visual stimulation, reward delivery, and licking behavior. Perievent time histograms revealed distinct temporal dynamics and magnitude changes in acetylcholine and calcium release in response to these stimuli. This cortex-wide imaging approach provides valuable insights into the spatiotemporal patterns of neuromodulatory signaling in primary sensory cortices, paving the way for further investigations into the role of these systems in modulating cortical responses, particularly in the context of Alzheimer's disease model mice.