Picture from Guha, Sanjib et al. (2020) MicroPubl Biol

Activation of the C. elegans mitochondrial unfolded protein response (UPRmt) in the PLM touch neuron by a tau mutant mimicking phosphorylation at T231: (A) A list of the strains used in this study. All transgenes code for translational fusions between TauT4 and the photo-convertible protein Dendra2, driven by the mec-7 promoter specifically in touch neurons. CRISPR-Cas9 gene editing was used to introduce phospho-ablation T231A, phospho-mimetic T231E and acetyl-mimetic K274/281Q mutations into the TauT4 ORF. For simplicity, these mutants will be referred to as T231A, T231E and K274/281Q. (B, C) Fluorescent images of a touch neuron from day 3 adult worms expressing single-copy transgenes coding for a Dendra2::TauT4 translational fusion and the T231E mutant. Dashed circles indicate the location of the PLM cell body, shown magnified in the inset. Scale bar, 0.5 um. Note, the blob fluorescence is coming from the hsp-60 expression tagged with GFP in the posterior intestine. (C, D) Quantification of the PLM cell body fluorescence for strains listed in panel A, at days 3 and 10 of adulthood. Data are the mean +/- SD from two independent technical replicates. Individual data points demarcate values from single PLM cells from separate animals (N = 25 +/- 5). Statistical analysis was by two-way ANOVA with Tukey's post hoc test, with *** P 0.001 when comparing bracketed samples. Note, the left side bar columns refer to the quantification of the fluorescence coming from the transgenic strains carrying Dendra2 reporter alone, whereas the right side ones refer to the strains carrying both Dendra2 and hsp-60 reporters. (E) Representative fluorescent images of transgenic worms expressing the integrated UPRmt reporter Phsp-60::GFP and single-copy MosSCI insertions containing either wild type TauT4 or T231E. Scale bar, 0.5 mm. (F) Quantification of the fluorescence signal intensity in the posterior intestinal region from the strains listed in panel A. Data are the mean +/- SD (N=20 animals from two independent biological replicates). ns denotes not significant, as calculated via one-way ANOVA followed by Tukey's multiple comparisons test.

Picture from Guha, Sanjib et al. (2021) MicroPubl Biol

Figure 1. Antixiodant MitoQ rescue of AWB impairment in pmp-4 mutant worms: A. Representative DIC-Nomarski picture of an L4 worm head. B-F: DiI staining. The DiI stained neurons is delimited by the black square. B. Wild type animal in which all neurons stained by the DiI are visible and named. C. DiI staining of the pmp-4(ok396) mutant animal. The white dashed circle indicates limited staining of the AWB neuron D. DiI staining of the rescue strain EDC010 pmp- 4(ok396);ibbEx42[Ppmp-4::pmp-4::gfp]. The white dashed circle indicates the faint staining of the AWB neuron. E, F. Wild type and pmp-4(ok396) worms treated with MitoQ. Scale bar, 0.5 um. G. Bar graph showing the worm fraction in which AWB neuron is visible after DiI staining, untreated and treated with MitoQ, of the corresponding genotypes (Fisher' test *** p<0.001, **p<0.01; n = 30 animals per genotype). H. Confocal image (GFP channel) of the reporter strain CX3877 kyIs156[Pstr-1::odr-10::gfp] showing cilia of AWB neuron. I, J. Representative confocal fluorescent pictures of the AWB cilia of the pmp-4(ok396) worm, untreated and treated with MitoQ. Scale bar, 0.1 um. K, L. Graph indicates the length of the long and short branch respectively expressed in um, of the indicated genotypes, treated and untreated with MitoQ. N= 40 cilia analyzed per genotype. One way ANOVA, Tukey's post hoc *** p<0.001, **p<0.01, *p<0.05). M Bar graph indicates odortaxis assay for wild type and pmp-4 mutant worms, at baseline and treated with MitoQ. One way ANOVA, Tukey's post hoc *** p<0.001, **p<0.01, *p<0.05). Each assay was conducted three times. ns, not significant.