Fig. 3
From: C-terminal calcium binding of α-synuclein modulates synaptic vesicle interaction
Alpha-synuclein and calcium balance the interaction of synaptic vesicles. a STED super-resolution imaging of isolated synaptic vesicles incubated with 1 mM EGTA, 200 µM calcium, 50 µM alpha-synuclein + 200 µM calcium, or 50 µM alpha-synuclein + 1 mM EGTA. Images show synaptic vesicles circled in red as detected for analysis of synaptic vesicle clustering. Scales represent 200 nm. Synaptic vesicle clustering is shown as a decrease of synaptic vesicles found as single vesicles and as an increase of synaptic vesicles found in clusters was seen upon incubation of synaptic vesicles with either increased calcium or alpha-synuclein. Note, EGTA was not able to reduce synaptic vesicle clustering in the presence of increased alpha-synuclein concentrations. **p = 0.0023, ****p < 0.0001, and **p = 0.0012 for comparison of % of single vesicles, *p = 0.0140, ***p = 0.0001, and *p = 0.0136 for comparison of % of vesicle clusters of two. Calculated using two-way ANOVA with Tukey’s post-hoc correction, graphs indicate mean ± s.e.m. N = 18 for all conditions, data from three biological repeats, d.f. 272. b TEM images of synaptic vesicles showing synaptic vesicle clustering in the presence of 50 µM alpha-synuclein. c Combined imaging of synaptic vesicles (confocal) and ATTO-647N-labeled alpha-synuclein (STED) showing synaptic vesicles surrounded and glued together by alpha-synuclein. Two biological repeats
