Figure 3. Ellipsoid body ring neurons express Sema-1a.
(A and B) Antibody staining reveals that Sema-1a and PlexA are both expressed in the EB during early pupal stages (24 hr and 48 hr APF). Sema-1a is also detected in the bulb (arrows) at 48 hr APF. (C) Schematic (modified from: Pecot et al., 2013) shows our strategy for using cell-type-specific flipase (FLP) expression to conditionally tag endogenous Sema-1a with V5 epitopes, revealing Sema-1a expression while at the same time conditionally labeling these same neurons with CD2-GFP. (D–F) Both CD2-GFP and V5-tagged Sema-1a were conditionally expressed in R neurons using the strategy outlined in panel C when R neuron-specific R11F03-GAL4 was used to express FLP. Low-magnification images show that CD2-GFP and V5 were not detected in animals when R11F03-GAL4 was not present (D). High-magnification images reveal that V5-tagged Sema-1a is specifically enriched in R neuron axonal terminals within the EB at 24 hr APF (E) and at 48 hr APF, unlike CD2-GFP, which is uniformly localized in both R neuron soma and axons (F). Sema-1a-V5 exhibits strong expression throughout the EB, and it is also found in R neuron dendritic terminals in the bulb at 48 hr APF. (G) Schematic (modified from: Pecot et al., 2013) showing how Sema-1a-expressing R neurons are labeled by lexA-driving mCherry when FLP is expressed in these same R neurons using GAL4 drivers. (H–J) Using the strategy in panel G, multiple GAL4 drivers were used to label different groups of R neurons with CD8-GFP. Sema-1a-expressing R neurons, identified by mCherry expression, are shown at 48 hr APF. Scale bars are 50 μm.
Figure 3—figure supplement 1. Sema-1a and PlexA are expressed in the central complex.
