Figure 4.

Drosophila TDP-43 ALS model. (A–F) Eyes from animals expressing GMR-GAL4 control (A) compared to GMR-dTDP-43^WT (B) and GMR-dTDP-43^mNLS (C). Expression of each construct results in ommatidia loss and a glossy appearance. GMR-GAL4-driven expression of dTDP-43^N493D results in larval lethality. (D–F) Cytoplasmic aggregates are observed in eye discs from individuals with GMR-GAL4-driven (D) wild-type dTDP-43^WT, (E) dTDP-43^N493D, and (F) dTDP-43^mNLS and stained with an antibody directed against human TDP-43 (red); nuclei are stained with DAPI (blue). We note that in control animals relatively low levels of endogenous dTDP-43 expression was detected using this antibody (Figure S2A). (G–O) OK371-GAL4 drives expression of UAS containing transgenic constructs in larval motor neurons; the OK371-GAL4 driver strain carries UAS-CD8-GFP, which labels the cell membrane of transgene-expressing cells with GFP (green). As above, ectopic dTDP-43 protein is detected with anti-TDP-43 (red) and nuclei are stained with DAPI (blue). (G–I) UAS-dTDP-43^WT and (J–L) UAS-dTDP-43^N493D show large cytoplasmic aggregated forms of dTDP-43, with little evidence of more broadly distributed cytoplasmic distributions, whereas (M–O) UAS-dTDP-43^mNLS exhibits dTDP-43 cytoplasmic mislocalization but appears to produce fewer aggregates and a more diffuse cytoplasmic pattern.