Fig. 4: TDP-43 protein mislocalizes in HD patient brain. | Nature Neuroscience

Fig. 4: TDP-43 protein mislocalizes in HD patient brain.

From: Aberrant splicing in Huntington’s disease accompanies disrupted TDP-43 activity and altered m6A RNA modification

Fig. 4

a, Representative IF staining images of SFG from patients with HD compared to non-HD control individuals showing decreased TDP-43 (yellow) signal intensity. b, Left: quantification of decreased nuclear TDP-43 signal intensity; five representative images were taken at ×40 from five HD and two control individuals. A CellProfiler pipeline was created to identify larger nuclei (enriched for neurons) by DAPI staining. The average of TPD-43 nuclear signal was obtained by measuring the intensity signal within a mask defined by DAPI. Each cell’s mean nuclear TDP-43 intensity is plotted. One-way ANOVA was performed with multiple comparisons and resulted in significant changes between all HD versus control comparisons (data not shown). Numbers on top of each group indicate the number of cells plotted. Right: dot plot showing grouped data by genotype. Statistical significance was derived from unpaired two-tailed t-test between control versus HD (P < 0.0001, t = 31.41, d.f. = 1317, F = 10.33, 95% CI: −0.03850 to −0.03397). Red bar represents the median. c, Representative IF staining images of the motor cortex from a patient with ALS (positive control) compared to patients with HD, using antibodies against total TDP-43 (yellow), phosphorylated TDP-43 (purple) and nuclear stain DAPI. White arrowhead indicates pTDP-43 cytoplasmic aggregation; red arrowhead (HD1) indicates cytoplasmic aggregate verified by orthogonal view. d, IF images showing pTDP-43 AL bodies (yellow) within MAP2-positive neurons (white). e, Bar graph showing counts of DAPI, MAP2-positive and AL bodies. Data are presented as mean values ± s.e.m. Each bar is derived from five random ×20 images from each patient. C, control; HD, patient with Huntington’s disease. %AL bodies is the number of AL bodies per patient normalized to the total number of Map2-positive neurons. Statistical significance was determined by unpaired two-tailed t-test (number of neurons: P = 0.3655, t = 0.9593, d.f. = 8, F = 5.329, 95% CI: −48.51 to 20.01; AL bodies (%): P = 0.0146, t = 3.103, d.f. = 8, F = 44.539, 95% CI: 0.08172–0.5545). Experiments in a–e were repeated at least three times with similar results represented above.

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