Biphasic training for awake imaging using a dual-imaging system reveals neurovascular uncoupling and anesthesia effects in healthy mice

Motivation: Most of rodent-fMRI is acquired under anesthesia, to minimize motion and stress. However, anesthesia hinders mouse-to-human translatability, since most of human fMRI is conducted whilst awake. Goal(s): Here we aim to develop a biphasic protocol for conducting awake rodent-fMRI in mice and simultaneously recording mesoscopic calcium imaging data. Approach: The animals undergo a first training and a refresher a few weeks later. Brain function is analyzed in simultaneous fMRI and mesoscopic calcium imaging measurements. Results: Having a refresher training improves motion in the scanner. The two measures of brain function show interesting patterns over time, with partial agreement and some clear disagreement. Impact: Rodent-fMRI is typically done under anesthesia to minimize stress and motion. This limits mouse-human translatability (since humans are usually scanned awake). Here, we develop a biphasic approach to train mice to awake imaging, using simultaneous fMRI and Calcium imaging measures.