BAcTrace, a tool for retrograde tracing of neuronal circuits in Drosophila

Animal behavior is encoded in neuronal circuits in the brain. To elucidate the function of these circuits, it is necessary to identify, record from and manipulate networks of connected neurons. Here we present BAcTrace (Botulinum Activated Tracer), a genetically encoded, retro-grade, transsynaptic labelling system. BAcTrace is based on C. botulinum neurotoxin A, Botox, which we have engineered to travel retrogradely between neurons to activate an otherwise silent transcription factor. We validate BAcTrace at three neuronal connections in the Drosophila olfactory system. We show that BAcTrace-mediated labeling allows electrophysiological recordings of connected neurons. Finally, in a challenging circuit with highly divergent connections, BAcTrace correctly identifies 12 out of 16 connections, which were previously observed by electron microscopy.