Transgenic tRNA^Gly overexpression rescues peripheral neuropathy phenotypes induced by hGlyRS-S211F and hGlyRS-H418R expression. (Aand B), Negative geotaxis climbing speed of 7-day-old female flies expressing hGlyRS-S211F or hGlyRS-H418R transgenes in motor neurons (OK371-GAL4), in the presence or absence of tRNA^Gly-GCC BAC (A) or tRNA^Gly-GCC scramble (B) transgenes. n = 7–24 (A) or 8–23 (B) groups of 10 flies per genotype; ***P< 0.0005 by Brown-Forsythe and Welch ANOVA with Dunnett's T3 multiple comparisons test (A) or one-way ANOVA with Sidak's multiple comparisons test (B). (C) Synapse length on distal muscle 8 of larvae expressing hGlyRS-S211F or hGlyRS-H418R transgenes in motor neurons (OK371-GAL4), in the presence or absence of tRNA^Gly-GCC overexpression. n = 9–12 animals per genotype; *P< 0.05, **P< 0.01, ***P< 0.001 by Brown-Forsythe and Welch ANOVA with Dunnett’s T3 multiple comparisons test. (D), Quantification of dendritic coverage (% of driver-only control) of class IV multidendritic sensory neurons in the larval body wall upon selective expression of hGlyRS-S211F or hGlyRS-H418R transgenes in these neurons (ppk-GAL4), in the presence or absence of tRNA^Gly-GCC overexpression. n = 12–19 animals per genotype; ***P< 0.0001 by one-way ANOVA with Sidak's multiple comparisons test. (Eand F), Quantification (E) and representative images (F) of relative translation rate (% of driver-only control) as determined by THRONCAT in motor neurons (OK371-GAL4) of larvae expressing hGlyRS-S211F or hGlyRS-H418R, in the presence or absence of tRNA^Gly-GCC overexpression. Motor neurons targeted by OK371-GAL4 are delineated by a dashed line and nuclei of motor neurons are labeled by a white dot (F). n = 7–12 animals per genotype; *P< 0.05, **P< 0.01, ***P< 0.001 by one-way ANOVA with Sidak’s multiple comparisons test. Scale bar: 10 μm.