During navigation, responses in primary visual cortex (V1) are modulated by the animal's position. Here, we show that this modulation is present across multiple higher visual cortical areas but largely absent in geniculate inputs to V1. Spatial modulation is stronger during active navigation than during passive viewing. Moreover, navigation activates different neurons than classical drifting gratings, and promotes the reliability of neural responses in parietal and medial cortical areas. Fig. 1: Spatial position modulates visual activity in visual cortex but not in thalamic LGN. a. Merged confocal images of somata in V1 (Nissl staining; blue) and of LGN axonal projections expressing GCaMP (GFP; green).GCaMP expression is densest in layer IV (L4). b. Example retinotopic map of the cortical surface with recording sessions targeting (fully or partly) V1 (black squares, field of view: 500µm x 500 µm). Black contours show borders between areas. c. Same retinotopic map as in b. with recording sessions targeting (fully or partly) higher visual areas (black squares, field of view: 500µm x 500 µm). d. Head-restrained mice traversed a virtual corridor by running on a Styrofoam wheel without a specific task. The corridor had two landmarks that repeated after 40 cm, creating visually matching segments. e. LGN response profiles as a function of position chosen from the 25 th , 50 th and 75 th percentile of the metric defined in h (top) and response profiles across the population (bottom). Response profiles are obtained from even trials and are normalized and ordered by the position of the maximum response estimated from odd trials. Only response profiles with variance explained ≥ 5% are included (1,140 of 3,182 synapses). Dotted lines are predictions assuming identical responses in the visually matching segments. f. Same as in e. for response profiles of V1 neurons (4,602 of 16,238 V1 neurons with variance explained ≥ 5%). g. Same as in e. and f. for response profiles of neurons across all 6 higher visual areas (4,381 of 18,142 neurons with variance explained ≥ 5%). h. Cumulative distribution of the spatial modulation index across visual areas (SMI) in even trials: SMI = 1 means single-peaked response; SMI = 0 means double-peaked response. Black: LGN, Gray: V1; Purple: mean cumulative distribution across higher visual areas (HVAs). Only neurons with responses within the visually-matching segments are included (LGN: 840/1,140, V1: 2,602/ 4,602, higher visual areas: 2,384/4,381) i. Violin plots showing the SMI distribution and median SMI (white vertical line) for each visual area (median ± m.a.d.