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Fig. 5. Study of Robo family members in sensory axon patterning. (A) Dorsal view of ppk-eGFP pattern in robo, robo2 and robo3 mutants show abnormal commissure formation or longitudinal branching. (B) Wild-type class IV axon scaffold in dorsal view (left) and Robo3 misexpression under the control of the pan-da neuron driver Gal4^109(2)80 (right). Robo3 induces ectopic collateral branch formation at the VL fascicle (arrows) and loss of some midline crossing by class IV neurons. Neurons are visualized using ppk-eGFP.(C) Robo2 misexpression in single neurons using MARCM does not cause disruptions in axon terminal layering or axon morphology (left panels). Shown are two class III neurons that terminate immediately next to the class IV axon scaffold, and two class II neurons that are separated from the class IV scaffold by a narrow space (arrows point to the collateral branches of the class II neurons). Robo3 misexpression causes defects in axon morphology in class II, III and IV neurons (right panels). The class II neuron ldaA truncates its arbor before reaching its usual position (arrow). Class III and class IV neurons show ectopic collateral branching (small arrowheads). The class IV neuron v'ada reaches a medial position comparable to wild-type class IV axons, but projects laterally in the next anterior segment when forced to express Robo3 (large arrowhead). Genotypes: (B) yw;Gal4^109(2)80/+ or Gal4^109(2)80/UAS-robo3; ppk-eGFP/+; (C) yw/hsFLP; UAS-robo2 or robo3/Gal4^109(2)80, UAS-mCD8::GFP; ppk-EGFP th st FRT2A/tubP-Gal80, FRT2A. Scale bars: 25 µm in B,C.

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