Figure 4
From: Novel animal model defines genetic contributions for neuron-to-neuron transfer of α-synuclein
Neuron-to-neuron transfer of a-syn requires synaptic transmission and is increased by disruption of autophagy or PD-associated genes. (A) Quantification of BiFC-induced fluorescence in BiFC-syn (WT) worms compared to BiFC-syn worms crossed with unc-11 and tom-1 mutant strains. (B) Quantification of BiFC-induced fluorescence in BiFC-syn worms crossed into the neuronal RNAi sensitive TU3401 strain following RNAi by feeding on HT115 bacteria expressing dsRNA of the respective autophagy related genes of interest. EV: Empty Vector control (light grey), DJR-1.1: off-target control, C. elegans isoform of DJR-1 expressed primarily in intestine (dark grey). (C) Quantification of BiFC-induced fluorescence in the head region of the worm of BiFC-syn worms following rapamycin treatment. (D) Lifespan of N2 and BiFC-syn worms following rapamycin treatment and without. (E) Quantification of BiFC-induced fluorescence in BiFC-syn worms crossed into the neuronal RNAi sensitive TU3401 strain following RNAi by feeding on HT115 bacteria expressing dsRNA of the respective autophagy related genes of interest. EV: Empty Vector control (light grey), DJR-1.1: off-target control. (G) Quantification of BiFC-induced fluorescence in the head region of the worm of BiFC-syn worms following bafilomycin treatment. (A–C,E,G) Statistical significance was determined from the mean values of three independent experiments with at least 75 worms per group, using ANOVA in conjunction with a Tukey’s post-hoc multiple comparisons test (alpha = 0.05). (F) mRNA expression levels of genes following RNAi knockdown. Statistical significance was determined from the mean values of three independent qPCR experiments using ANOVA in conjunction with a Tukey’s post-hoc multiple comparisons test (alpha = 0.05). In all cases data is represented as mean of 3 independent experiment, represented as mean ± SEM.
