Fig. 1
Characterisation of oligomers and their effect on mitochondria. a Representative SAVE images of early oligomers (4 h), late oligomers (8 h), and fibrils (24 h). Zoom and representative two-dimensional Gaussian distribution fits are shown in the insets. The scale bar is 5 µm and 1 µm in the zoom, and the colour bar shows the Gaussian amplitude (×104 photons). b Quantification of aggregation. Each detected species was fitted to a two-dimensional Gaussian distribution, and histograms of the widths (FWHM) along the longest axis, and the total integrated intensities are shown for each time-point. c Representative traces from single experiments, of NADH autofluorescence in WT neurons exposed to either monomers (n = 15 cells), oligomers (n = 32 cells), or oligomers ± CsA (n = 9 cells and n = 6 cells, respectively). d Representative Rh123 traces of cells challenged with oligomers. e Quantification of mitochondrial depolarisation upon either monomeric or oligomeric application. N = 3 experiments; Oligomers: n ≥ 60 neurons/astrocytes; Monomers: n ≥ 50 neurons/astrocytes. f Representative Rh123 traces of cells challenged with oligomers. g Representative Rh123 traces of cells challenged with oligomers ± CsA. h Representative traces of Rh123 and fura-2 in WT neurons exposed to glutamate in the presence of monomers or oligomers. i Representative images of WT rat neurons labelled with fluo-4 (cytosolic calcium) and TMRM (ΔΨm) before (0 min) and after (4 min) high laser exposure which represents PTP opening. A representative trace of TMRM and Fluo-4 fluorescence where the drop in TMRM fluorescence precedes increase in Fluo-4 fluorescence. Quantification of the time until PTP opening in WT cells pre-exposed to oligomeric α-synuclein. N = 3 experiments; n = 43 cells for control and n = 20 cells for oligomers. Two-tailed Student’s t-test for e and i. Scatter points represent individual cells for e and i. Scale bar = 10 μm. Data represented as mean ± SEM. *p < 0.05; **p < 0.01; ***p < 0.001
