Inhibitory circuits coordinate leg movements during Drosophila grooming

Limbs execute diverse actions coordinated by the nervous system through multiple motor programs. The basic architecture of motor neurons that activate muscles which articulate joints for antagonistic flexion and extension movements is conserved from flies to vertebrates. While excitatory premotor circuits are expected to establish sets of leg motor neurons that work together, our study uncovered an instructive role for inhibitory circuits. Using electron microscopy data for the Drosophila nerve cord, we categorized ∼120 GABAergic inhibitory neurons from the 13A and 13B hemi-lineages into classes based on similarities in morphology and connectivity. By mapping their synaptic partners, we uncovered redundant pathways for inhibiting specific groups of motor neurons, disinhibiting antagonistic counterparts, or inducing alternation between flexion and extension. We tested the function of specific inhibitory neurons through optogenetic activation and silencing, using quantitative leg movement assays for coordination during grooming. Behavior experiments and modeling demonstrate that inhibition can induce rhythmic motion, highlighting the importance of inhibitory circuits in motor control.