The Drosophila connectome reveals Axo-Axonic Synapses on Descending Neurons that modulate the Giant Fiber System

Abstract Axo-axonic synapses can veto, amplify, or synchronize spikes, yet their circuit-scale logic is unknown. Using the complete electron-microscopy connectome of the adult male Drosophila, we charted every axo-axonic input onto the 1,314 descending neurons that carry brain commands to the ventral nerve cord. By definition, any synapse connected to a descending neuron within the cord is axo-axonic. Thus, we uncovered the ascending-descending and interneurons-descending axo-axonic relationship. Neurons with many partners (high-degree) integrate into the network without clustering into an interconnected ‘rich-club’ of hubs. We identified an octet of ascending neurons (AN08B098) whose axo-axonic input to the Giant Fibers (DNp01) predicted modulation of the escape circuit. Immunostaining confirms their cholinergic identity, while optogenetic activation confirmed that this excitatory cohort increases DNp01 excitability, validating connectome-derived rules. Our work delivers a map of axo-axonic wiring in a complete ventral nerve cord connectome and provides constraints for models of fast motor control. Competing Interest Statement The authors have declared no competing interest. Footnotes Methos are now in STARs format. Limitations to the study section was added.