Connectome simulations identify a central pattern generator circuit for fly walking

Abstract Animal locomotion relies on rhythmic body movements driven by central pattern generators (CPGs): neural circuits that produce oscillating output without oscillating input. However, the circuit structure of a CPG for walking is not known in any animal. To identify the cells and synapses that underlie rhythmic leg movement in walking flies, we developed dynamic simulations of the Drosophila ventral nerve cord (VNC) connectomes. A computational activation screen of descending neurons from the central brain identified DNg100—a known command neuron for walking—as the top driver of rhythmic leg motor activity. Simulated network pruning isolated a minimal rhythm-generating circuit consisting of one inhibitory and two excitatory interneurons; this three-neuron circuit was necessary and sufficient for motor rhythms across all six legs and in four connectome datasets. Simulations also predicted that a separate descending pathway (DNb08) drives rhythmic leg movements, which we confirmed experimentally using optogenetics in behaving flies. Our results reveal the cellular identity and synaptic structure of a putative CPG circuit for walking and other rhythmic leg movements in flies. Competing Interest Statement The authors have declared no competing interest. Footnotes We have made substantial revisions to the manuscript that fall under three different umbrellas: I. Robustness of simulation results across model parameters II. In-depth analyses of the structure of the putative core CPG circuit III. Replication of motor rhythms in 2 newly published fly CNS connectome datasets and in all legs IV: Accessibility of code repository