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. 2021 Mar 5;12:1484. doi: 10.1038/s41467-021-21611-8

Fig. 1. Blunt force trauma induces neuronal dysfunction in C. elegans.

Fig. 1

a Diagram depicting method of delivering blunt trauma to worms. b Micrographs of worms exhibiting movement with normal body bend (left) versus post injury paralysis (right). Reference Fig. 1c. Scale = 400 µm. c Percent of animals entering paralytic state immediately post injury is dependent on agitation frequency. Mean ± SEM, ns (not significant) p = 0.7044 and 0.4494, ****p < 0.0001 by two-way ANOVA with Tukey multiple comparison test. n = 3 independent trials. d Fluorescence maximum projections of Z-stacks taken by confocal microscopy of worms expressing GFP in GABAergic neurons. Worms imaged either uninjured or 1 h post injury. Reference Supplementary Fig. 1b. Scale = 100 µm. e Paralytic response to injury in either a control worm strain (from Fig. 1c, black circles), or worms expressing polyglutamine expansions Q40::YFP (soluble, green squares) or Q67::YFP (insoluble, red triangles) in neurons. Mean ± SEM, ****p < 0.0001, ***p = 0.0001 by two-way ANOVA with Tukey test, n = 3 independent trials. f Filter trap analysis of SDS-resistant aggregates on cellulose acetate membranes from neuronally expressed Q40::YFP worms either uninjured or 48 h post injury. Western blots for the respective proteins resolved by SDS-PAGE in the bottom panel. g Quantification of filter trap. Shown are log-transformed values of slot blot band intensity with black and blue denoting uninjured and injured worms, respectively (corrected for loading by YFP signal). Mean ± SEM, **p = 0.0008 by two-sided ratio-paired t-test, n = 3 biological replicates. h Micrographs of worm tracts on E. coli lawn to assess movement 48 h after receiving no injury, non-paralytic mild injury (7500 rpm), or paralytic moderate injury (8600 rpm). Scale = 1 mm. i Chemotaxis of uninjured (black circles) or injured worms (blue squares) with butanol 24–96 h post injury. Chemotactic index of 1.0 indicates 100% preference for butanol and 0.0 indicates no preference. Mean ± SEM, ****p < 0.0001 and uninjured Day 1 vs Day 4 ***p = 0.0005 by two-way ANOVA with Tukey test, n = 6 independent trials for day 1, 2, 4 and n = 3 independent trials for day 3. j Gentle nose touch assay of uninjured (black circles) or injured worms (blue squares), in which a stop and reversal of worm movement indicated a positive response. Mean ± SEM, ***p = 0.0006 and 0.0004, ****p < 0.0001 when compared to uninjured by two-way ANOVA with Tukey test, n = 200 animals per condition over three independent trials.