(A) Diagram of the three groups of mice reared with different visuomotor experience. Normally reared mice (NR, gray) were raised with a normal light/dark cycle in their cages, with the full diversity of visuomotor coupling in freely moving conditions. Coupled trained mice (CT, blue) were reared in the dark, and their only visuomotor experience prior to imaging was during closed loop conditions in the virtual reality system. Non-coupled trained mice (NT, orange) were reared similar to coupled trained mice, except their only visual experience was during open loop conditions in the virtual reality system. (B) Average population responses of layer 2/3 neurons in V1 to the onset of visual flow during open loop replay sessions. Green shows the average speed profile of the visual flow stimulus, purple shows the average population responses during locomotion, and black shows the average population responses during stationary periods for normally reared (top) and coupled trained (bottom) mice. Shading indicates SEM. (C) Distribution of the locomotion modulation indices of all V1 layer 2/3 neurons for the visual flow onset responses recorded in normally reared (gray), coupled trained (blue), and non-coupled trained (orange) mice. Here and in other panels, n.s.: not significant, *: p<0.05, **: p<0.01, ***: p < 0.001. For complete statistical information see Supplementary file 1. (D) Average population responses of layer 2/3 neurons in V1 in coupled trained mice to the onset of visual flow during stationary periods (black), and during slow (top) and fast (bottom) locomotion (purple). Shading indicates SEM. (E) Comparison of locomotion modulation indices averaged across neurons for visual flow onset responses recorded during slow, medium, and fast locomotion speeds, in normally reared (left) and coupled trained (right) mice. Error bars indicate SEM. Green dashed line indicates linear regression fit to the data, and shading indicates the 95% confidence interval of the regression. Green asterisks and ‘n.s.’ indicate significance of the fit. (F) As for panel D, but for nasotemporal visual flow responses in high ChrimsonR-expressing mice after LC axon stimulation in closed loop conditions. (G) As for panel E, but for nasotemporal visual flow responses in high ChrimsonR-expressing mice before (left) and after (right) LC axon stimulation in closed loop visuomotor coupling.