Independent validation of transgenerational inheritance of learned pathogen avoidance in Caenorhabditis elegans
- School of Biological Sciences, Illinois State University, United States
Figures
Figure 1
Transgenerational inheritance of PA14 avoidance in C. elegans.
(A) Chemotaxis assay setup. A 25 µL drop of OP50 and PA14 bacteria of the same optical density (OD600=1) plus 1.0 µL of 400 mM NaN3 were placed equidistant from the center of a 10 cm chemotaxis plate where worms were placed at the start of the assay (see Materials and methods). Indices were measured for naïve worms, trained worms, first-generation (F1), and second-generation (F2) offspring. (B) Pooled results from four independent trials showing that trained worms, which were exposed to PA14 for 24 hr, exhibited significantly higher choice indices, indicating learned avoidance of PA14. This aversion persisted in F1 and F2 progeny, demonstrating transgenerational inheritance of pathogen avoidance behavior. (C) Comparison of data separated into individual trials. Each point is one plate assay. Bars show mean ± s.e.m. Sample sizes (assays): Naïve n = 38, Trained n = 38, F1 n = 19, F2 n = 43. Datasets that met normality and equal variance were analyzed by one-way ANOVA with Holm–Sidak post hoc comparisons versus Naïve; otherwise Kruskal–Wallis with Dunn’s post hoc was used. Significance versus Naïve is indicated as *p < 0.05 or **p < 0.01, as reported in Supplementary file (see Supplementary file 1A for statistics and Supplementary file 1B for raw data).
Additional files
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Supplementary file 1
Full statistics, raw chemotaxis assay counts, and reagent lists for all experiments.
(A) Statistical analyses of chemotaxis choice indices across generations. Results from Kruskal-Wallis one-way analysis of variance (ANOVA) followed by Dunn’s or Holm-Sidak post hoc comparisons are reported for naïve, trained, F1, and F2 animals. For each trial, test statistics (H, DF, p values) and pairwise comparisons are presented. Normality (Shapiro-Wilk) and equal variance (Brown-Forsythe) test outcomes are included. (B) Raw chemotaxis assay data for all replicates. Counts of worms immobilized on PA14 and OP50 spots are listed for each assay, along with calculated choice index (CI) values. Data are separated by generation (naïve, trained, F1, F2) and trial number, providing the full dataset underlying Figure 1. (C) Reagents used in this study compared with those supplied by the Murphy and Hunter groups. The table lists reagents, suppliers, and catalog numbers for nematode growth medium (NGM) ingredients and related chemicals. Parallel listing highlights methodological consistencies and differences across the three laboratories.
- https://cdn.elifesciences.org/articles/107034/elife-107034-supp1-v1.xlsx
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MDAR checklist
- https://cdn.elifesciences.org/articles/107034/elife-107034-mdarchecklist1-v1.docx
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Independent validation of transgenerational inheritance of learned pathogen avoidance in Caenorhabditis elegans
eLife 14:RP107034.
https://doi.org/10.7554/eLife.107034.3
