Fig. 1: Despite lifelong p62 accumulations, spinal cord motor neurons in mice are resistant to Tbk1 loss. | Nature Communications

Fig. 1: Despite lifelong p62 accumulations, spinal cord motor neurons in mice are resistant to Tbk1 loss.

From: ALS/FTD-linked TBK1 deficiency in microglia induces an aged-like microglial signature and drives social recognition deficits in mice

Fig. 1

a Quantification of motor neurons (MN) in the lumbar spinal cord of 20-month-old mice with MN specific full Tbk1 deletion (Chat-Cre/Tbk1fl/fl, hereafter Tbk1-MN-KO) compared to littermate control (Tbk1-MN-WT) mice. n = 4 mice per genotype (equally sex-mixed). Adjusted p value is indicated and was determined using a two-tailed unpaired t-test. b Representative confocal immunofluorescence images of the autophagy receptor p62 and the MN marker ChAT in lumbar spinal cord sections from 3-month-old Tbk1-MN-KO and Tbk1-MN-WT mice, showing MN with p62+ accumulations (puncta) only in Tbk1-MN-KO mice (arrowheads). Scale bar = 50 µm, scale bar in enlargements 10 µm. c Quantification of MN containing p62+ accumulations at 3 months (13.6%) and 20 months (26.1%). n = 4 mice per genotype and age (equally sex-mixed). Adjusted p values are indicated and were determined using a two-way ANOVA with uncorrected Fisher LSD. d Representative brightfield images of laser-microdissection of MN (for RNAseq), identified by Nissl staining, from mouse lumbar spinal cord sections. Scale bar = 100 µm. e Volcano plot and bar graph of significantly deregulated genes (DEGs) (DESeq2, log2FC ≥ 0.5, adjusted p values (FDR) < 0.05, see “Methods”) identified by RNAseq from laser-microdissected MN (pools of 200 MN per mouse), between 20-month-old Tbk1-MN-KO and Tbk1-MN-WT mice, showing beside expected Tbk1 downregulation only very few DEGs. n = 6 mice per genotype (females only). FC fold change, p.adj adjusted p value, UP upregulated, DOWN downregulated, nc not changed. f Acute MN injury, using the unilateral sciatic nerve crush/regeneration paradigm, was performed in 6-month-old Tbk1-MN-KO and Tbk1-MN-WT mice. Recovery of the sciatic functional index (SFI) was calculated based on toe-spread and paw print length, over 4 weeks after crush. n = 7–8 mice per genotype (females only). Adjusted p values were determined using repeated measures one-way ANOVA with Sidak’s multiple comparisons test. a, c Data are shown as single data points and (a, c, f) means ± SEM. ns not significant.

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