Fig. 3: Sleep history alters mitochondrial dynamics.
a,b, Volumetric renderings (a) and morphometric parameters (b) of automatically detected mitochondria in OPRM image stacks of dFBN dendrites in rested flies, sleep-deprived flies and flies allowed to recover for 24 h after sleep deprivation. Sleep history-dependent changes in mitochondrial number (P < 0.0001, Holm–Šídák test after ANOVA), volume (P = 0.0470, Dunn’s test after Kruskal–Wallis ANOVA), sphericity (P = 0.0124, Dunn’s test after Kruskal–Wallis ANOVA) and branch length (P = 0.0033, Dunn’s test after Kruskal–Wallis ANOVA) are occluded by the co-expression of AOX (P ≥ 0.2257, two-sided t-test or Mann–Whitney test) or the simultaneous activation of TrpA1 (P ≥ 0.0625, two-sided t-test or Mann–Whitney test) and (over)corrected after recovery sleep (number of mitochondria: P = 0.1551, all other parameters: P ≤ 0.0302, Dunn’s test after Kruskal–Wallis ANOVA). Two data points exceeding the y-axis limits are plotted as triangles at the top of the graphs; mean and s.e.m. are based on the actual values. c, Drp1 recruitment. Single confocal image planes through dFBN somata of flies expressing R23E10-GAL4-driven mito-GFP (top) and Drp1Flag from the endogenous locus (bottom). Sleep deprivation increases the percentage of cellular anti-Flag fluorescence (intensity-coded according to the key below) within automatically detected mitochondrial contours (P < 0.0001, two-sided Mann–Whitney test). d, Mitochondria–endoplasmic reticulum contacts. Isosurface renderings (voxel value 128) of SPLICS puncta in distal dFBN dendritic branches (dashed outlines), obtained by trilinear interpolation of thresholded and despeckled confocal image stacks. Sleep deprivation increases the number of SPLICS puncta per dendritic field (P < 0.0001, two-sided Mann–Whitney test). e, Mitophagy. Summed-intensity projections of dFBN dendrites expressing mito-QC. Emission ratios are intensity-coded according to the key below and increase after sleep deprivation (P = 0.0101, two-sided t-test). Data are means ± s.e.m.; n, number of dendritic regions (b,d,e) or somata (c); asterisks, significant differences (P < 0.05) in planned pairwise comparisons. Scale bars, 10 μm (a), 2 μm (c), 10 μm (d), 5 μm (e). For statistical details see Supplementary Table 1.
