Figure 5. SIn_ACh Axons in ACtx are Active During Movements.

(A) Schematic of the experimental strategy. AAV-FLEX-GCaMP6s was injected into SIn of ChAT-Cre mice. (B) Confocal micrograph of GCaMP-labeled SIn neurons. Neurons expressing GCaMP expressed ChAT (red). (C) Confocal micrograph of GCaMP-expressing SIn_ACh axons in ACtx. (D) Outline of experimental design for 2 photon imaging and tracking movements. ROIs for monitoring body movements are indicated with dashed boxes. (E) Average fluorescence of a GCaMP-labeled axon, imaged in vivo. The region outlined with a green dashed line indicates the ROI for monitoring changes in fluorescence over time. (F) Pseudocolored axonal fluorescence during rest. (G) Pseudocolored axonal fluorescence during a body movement. (H) ∆F/F from a single ROI (blue trace) during movements and rest (red trace). The dashed line indicates the threshold for identifying movements. (I) ∆F/F during paw movements (n = 12 axons, 2 mice). (J) ∆F/F during mouth movements (n = 12 axons, 2 mice). (K) ∆F/F during all body movements (n = 14 axons, 3 mice). (**p < 0.01, *** p < 0.001). Values are mean ± SEM.