Drosophila Brainbow: a recombinase-based fluorescent labeling technique to subdivide neural expression patterns

We developed a multicolor neuron labeling technique in Drosophila melanogaster combining the power to specifically target different neural populations with the label diversity provided by stochastic color choice. This adaptation of vertebrate Brainbow uses recombination to select one of three epitope-tagged proteins detectable with immunofluorescence. Two copies of this construct yield six bright, separable colors. Here we use Drosophila Brainbow to show the innervation patterns of multiple antennal lobe projection neuron lineages in the same preparation and to observe the relative trajectories of individual aminergic neurons. Nerve bundles, and even individual neurites hundreds of microns long, can be followed with definitive color labeling. We trace motor neurons in the subesophageal ganglion and correlate them to neuromuscular junctions to show their specific proboscis muscle targets. The ability to independently visualize multiple lineage or neuron projections within the same preparation greatly advances the goal of mapping how neurons connect into circuits.