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. 2021 Apr 26;22(6):e50958. doi: 10.15252/embr.202050958

Figure 6. GABAergic defects and behavioural anomalies are suppressed by ephedrine treatment in mut‐chd7 Caenorhabditis elegans .

Figure 6

  • A
    Movement scores of chd‐7(gk290) mutants (red) compared with N2 wildtype (WT; black). Two‐tailed t‐test was performed for statistical analyses, and movement score was considered different to WT (N = 3, n = 30; ****P < 0.0001, Student’s t‐test).
  • B
    Lifespan analyses of chd‐7(gk290) mutants (n = 339; red) compared with WT (n = 444; black). Log‐rank test was performed for statistical analyses. ***P < 0.001.
  • C
    WT and chd‐7(gk290) animals with defects in the GABAergic (black) and cholinergic nervous system (grey) at the L4 stage (N = 4, n = 100; ****P < 0.0001; Student’s t‐test).
  • D–F
    Example pictures of the GABAergic nervous system cell bodies, commissures, axonal gaps and axonal breaks larger than 50 µm per worm in chd‐7(gk290) mutants expressing unc‐47p::mCherry.
  • G–I
    GABAergic neurodevelopmental defects of WT (black) and chd‐7(gk290) mutants (grey) at the L4 stage classified in axonal gaps per worm (G), number of GABAergic cell bodies (H) and number of commissures (I). N = 4, n = 100; ****P < 0.0001, Student’s t‐test.
  • J
    Chemical libraries (3,850 compounds) were first screen in chd‐7(gk290) C. elegans mutants and positive hits were tested on chd7 zebrafish mutants.
  • K
    49 compounds that improved motility phenotypes in chd‐7(gk290) mutants were identified. These compounds were clustered in 6 main functional categories.
  • L
    Movement scores of chd‐7(gk290) mutants treated with ephedrine (blue) compared with the solvent DMSO (black). N = 3, n > 100; ****P < 0.0001, Student’s t‐test.
  • M
    Defects of the GABAergic nervous system at the L4 stage in chd‐7(gk290) mutants treated with ephedrine (grey) compared with DMSO (black) (N = 3, n > 100). ****P < 0.0001, Student’s t‐test.
  • N
    Average activity per second is wild‐type and mutants treated with ephedrine compared with non‐treated mutant (wild‐type treated with ephedrine is referred to as Control). Ephedrine suppresses hyperactivity in chd7 mutant fish. n = 24; ****P < 0.0001, one‐way ANOVA.
  • O
    Representative images of GABAergic neurons and total number of GABAergic neurons (GFP+ cells) in the optic tectum (OT) and cerebellum (CB) regions of wild‐type, chd7 mutant and ephedrine‐treated fish (n = 9; ****P < 0.0001; One‐way ANOVA). Treatment with ephedrine ameliorated the number of GABAergic neurons.
  • P, Q
    pERK is reduced in mutants following treatment with ephedrine as shown by Western blot quantification (N = 4). ****P < 0.0001, one‐way ANOVA.
  • R
    qPCR analysis of paqr3b expression in chd7 +/+ , chd7 −/− and chd7 −/− treated with ephedrine (N = 4; ****P < 0.0001; ns, not significant, one‐way ANOVA).

Data information: Data are presented as mean ± SEM. Scale bar = 50 μm. n is the number of fish or worms used. N is the number of experimental repeats.