Flybow: genetic multicolor cell labeling for neural circuit analysis in Drosophila melanogaster

Nat Methods. 2011 Mar;8(3):260-6. doi: 10.1038/nmeth.1567. Epub 2011 Feb 6.


To facilitate studies of neural network architecture and formation, we generated three Drosophila melanogaster variants of the mouse Brainbow-2 system, called Flybow. Sequences encoding different membrane-tethered fluorescent proteins were arranged in pairs within cassettes flanked by recombination sites. Flybow combines the Gal4-upstream activating sequence binary system to regulate transgene expression and an inducible modified Flp-FRT system to drive inversions and excisions of cassettes. This provides spatial and temporal control over the stochastic expression of one of two or four reporters within one sample. Using the visual system, the embryonic nervous system and the wing imaginal disc, we show that Flybow in conjunction with specific Gal4 drivers can be used to visualize cell morphology with high resolution. Finally, we demonstrate that this labeling approach is compatible with available Flp-FRT-based techniques, such as mosaic analysis with a repressible cell marker; this could further support the genetic analysis of neural circuit assembly and function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • DNA-Binding Proteins / genetics
  • Drosophila melanogaster / cytology*
  • Drosophila melanogaster / embryology
  • Drosophila melanogaster / genetics
  • Luminescent Proteins / analysis*
  • Luminescent Proteins / genetics
  • Mice
  • Molecular Sequence Data
  • Nerve Net / cytology*
  • Nerve Net / embryology
  • Neuroglia / chemistry
  • Neuroglia / cytology
  • Neuroglia / metabolism
  • Neurons / chemistry
  • Neurons / cytology*
  • Neurons / metabolism
  • Saccharomyces cerevisiae Proteins / genetics
  • Staining and Labeling / methods*
  • Transcription Factors / genetics


  • DNA-Binding Proteins
  • GAL4 protein, S cerevisiae
  • Luminescent Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors

Associated data

  • GENBANK/HQ998853
  • GENBANK/HQ998854
  • GENBANK/HQ998855