Fig. 5: Neuronal expression of Synaptojanin rescues pathogenic Auxilin mutants.
From: Parkinsonism mutations in DNAJC6 cause lipid defects and neurodegeneration that are rescued by Synj1
a Quantification of negative geotaxis assay of 5DO dAuxWT/F956X and dAuxRG/F956X flies and of dAuxWT/F956X and dAuxRG/F956X flies that neuronally express wild type Drosophila Auxilin (UAS-AUX), harbor an extra genomic copy of wild type Drosophila Clathrin heavy chain (HA-CHC) or neuronally express wild type Drosophila Synaptojanin (UAS-Synj). Graphs represent the fraction of flies crossing the threshold. Bars show mean ± SEM. Points represent the mean of three trials. One-way ANOVA, Dunnett’s multiple comparison test. ns: not significant, *p < 0.05, ***p < 0.001 and ****p < 0.0001. b. Quantification of seizures following stimulation in 5DO flies of indicated genotypes (see also a). Note that dAux and dSynj rescue pathogenic dAux mutants, but dChc does not. Graphs represent the fraction of flies that were unable to stand after 10 s after stimulation. Bars show mean ± SEM. Points represent single trials with 10 flies each. One-way ANOVA, Dunnett’s multiple comparison test, n≥5. ns: not significant, ***p < 0.001 and ****p < 0.0001. c - e. Brightfield images of toluidine blue stained sections of 15DO dAuxWT/F956x and dAuxRG/F956X flies neuronally expressing wild type Drosophila Auxilin (UAS-AUX), harboring an extra genomic copy of wild type Drosophila Clathrin heavy chain (HA-CHC) or neuronally expressing wild type Drosophila Synaptojanin (UAS-Synj) (c) and quantification of the degenerated area per standardized section (d) as well as the amount of normal versus abnormal rhabdomeres (see Methods) as a fraction of the total number of rhabdomeres (e). Note that again, dAux and dSynj rescue pathogenic dAux mutants, but dChc does not. Asterisks indicate degeneration in the retina. Scale bar: 5 µm. Graph in d shows mean ± SEM of n≥5 flies per genotype (from independent crosses), points represent individual values. Kruskal-Wallis test, Dunnett’s multiple comparison test. ns: not significant, *p < 0.05 and **p < 0.01. Graph in e shows mean ± SD of n≥5 flies per genotype from independent crosses. Two-way ANOVA with Tukey’s multiple comparison test. ns: not significant, **p < 0.01, ***p < 0.001 and ****p < 0.0001. f. Necrotic spots disappear and rough eye phenotype improves by neuronal expression of Synaptojanin. Panel shows representative brightfield images of the compound eye of 5DO flies of indicated genotypes. Insets show higher magnification. Scale bars (up) 100 µm; (down) 20 µm.
