Mapping the neural dynamics of locomotion across the Drosophila brain

Locomotion engages widely distributed networks of neurons. However, our understanding of the spatial architecture and temporal dynamics of the networks that underpin walking remains incomplete. We use volumetric two-photon imaging to map neural activity associated with walking across the entire brain of Drosophila. We define spatially clustered neural signals selectively associated with changes in either forward or angular velocity, demonstrating that neurons with similar behavioral selectivity are clustered. These signals reveal distinct topographic maps in diverse brain regions involved in navigation, memory, sensory processing, and motor control, as well as regions not previously linked to locomotion. We identify temporal trajectories of neural activity that sweep across these maps, including signals that anticipate future movement, representing the sequential engagement of clusters with different behavioral specificities. Finally, we register these maps to a connectome and identify neural networks that we propose underlie the observed signals, setting a foundation for subsequent circuit dissection. Overall, our work suggests a spatiotemporal framework for the emergence and execution of complex walking maneuvers and links this brain-wide neural activity to single neurons and local circuits.