Figure 6.

dPLD effects on GMR-hFUS^R521C, GMR-hTDP-43^M337V, and GMR-c9orf72(G4C2)₃₀ phenotypes. (A–D) RNAi-induced dPld reduction suppressed GMR-hFUS^R521C and GMR-hTDP-43^M337V phenotypes. Representative eye images of individuals carrying one copy of (A) GMR-hTDP-43^M337V in trans with (B) UAS-dPld³²⁸³⁹, or (C) GMR-hFUSR^521C in trans with (D) UAS-dPld³²⁸³⁹. dPld³²⁸³⁹ expresses an RNAi directed against dPld. (E) Histogram representation of percentage of GMR-c9orf72(G4C2)₃₀ individuals displaying the degenerative phenotype at week 1 (blue), week 3 (orange), and week 6 (gray). GMR-GAL4-directed expression of UAS-dPld³²⁸³⁹ results in suppression of the c9orf72(G4C2)₃₀-dependent neurodegenerative phenotype at all three time points. (F–H) Effects of motoneuron-driven dPLD on third instar larval NMJ morphology. Representative images of third instar larval NMJs from (F) control OK371-GAL4 as well as loss [(G) UAS-dPld³²⁸³⁹] and gain [(H) UAS-dPld13] (Raghu et al. 2009) of dPld function in OK371-GAL4/+ background are shown. (I) Histogram representation of quantification of average bouton numbers per muscle in OK371-GAL4 (OK371), OK371-GAL4/+ UAS-dPld³²³⁸⁹/+ (dPLD.RNAi, gray), OK371-GAL4/UAS-dPld13 (UAS.dPLD, red), OK371-GAL4, UAS-dTDP-43^N493D (N493D, yellow), and OK371-GAL4, UAS-dTDP-43^N493D/UAS-dPld³²⁸³⁹ (N493D/dPLD.RNAi, green) individuals. Loss of dPld has no effect on OK371-GAL4 bouton number, whereas there is a statistically significant decrease (**** P < 0.0001) in bouton number when dPLD is ectopically expressed. Quantifications were done manually at the confocal microscope and statistical significance was determined by using an unpaired parametric t-test with Prism software. NMJ preparations were stained with anti-HRP (green) and anti-discs large (Dlg) (red) to mark pre- and postsynaptic structures, respectively, and muscle nuclei were labeled with DAPI. The more asterisks, the smaller the P-value.