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oai:figshare.com:article/34049539 2026-10-01T17:47:06Z category_4 category_12 category_13 category_14 category_15 category_16 category_64 category_128 portal_5 item_type_1 month_year_10_2026

egl-2(G574R) is a gain-of-function allele and disrupts EGF-induced sleep. Xinyu Huang (679541) Sudharsan Kannan (25158064) Gail A. Robertson (15007385) Han Wang (254423) Biochemistry Cell Biology Genetics Molecular Biology Neuroscience Physiology Cancer Hematology regulate neuronal excitability promoting neuron ala influence channel gating hallmark cyclic nucleotide genetic analysis demonstrated forward genetics reveals electrophysiological analysis showed dependent channel activation calcium imaging showed binding homology domains adjacent channel subunits accelerates activation kinetics subfamily kcnh channels conserved molecular mechanism important sleep regulator findings identify egl div >< p caenorhabditis elegans </ similarly suppresses sleep 2 functions cell elegans </ suppresses sleep kcnh voltage vivo </ central sleep conserved cnbhd shifts voltage second substitution negative potentials intrinsic ligand g574r ), function effects corresponding residue (A-B) Quantification of sleep behavior as the fraction of quiescent worms for the indicated genotypes and conditions. All strains contain the transgene (HS::EGF) for conditional overexpression of EGF via 15-min heat shock (HS) at 33 °C to induce sleep. Pre and post refer to before and 2 h after heat shock, respectively. egl-2(tan200) and egl-2(gk593448) were used as egl-2(G574E) and egl-2(G574R), respectively. Homozygous (A) and heterozygous (B) animals were assayed to compare the sleep phenotype. Each dot represents one independent assay containing 15–25 animals on a single plate. Mean ± SEM are shown. ns, not significant; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. One-way ANOVA with Tukey’s multiple-comparisons test for post-heat shock samples. (C) Time course of endogenous stress-induced sleep in animals of the indicated genotypes shown as the fraction of quiescent worms at each time point. Pre refers to before heat shock. The egl-2(gk583448) allele was used for egl-2(G574R). Data were collected from three independent replicates, with 15–25 animals of each strain assayed on separate plates in each replicate. Mean ± SEM are shown. ns, not significant; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001. Two-tailed unpaired Student’s t-test for comparing wild type and egl-2(gk583448) mutants at each time point. (D-E) Representative images and quantification of endogenous EGL-2 protein expression in the ALA neuron using a split-GFP strategy. (D) Top: egl-2(+)::7x gfp11; Middle: egl-2(G574E)::7x gfp11; bottom: egl-2(G574R)::7x gfp11 animals. All worm strains also carried the transgene ALAp::gfp1–10 for ALA-specific expression of GFP1–10. White dashed ovals outline the ALA neuron cell body. A, anterior; D, dorsal. Scale bar, 20 μm. (E) Numbers shown on the bars indicate the total number of animals imaged for each genotype. ns, not significant; *, p < 0.05; ****, p < 0.0001. One-way ANOVA with Tukey’s multiple-comparisons test. 2026-10-01T17:46:48Z Image Figure 10.1371/journal.pgen.1012255.g006 https://figshare.com/articles/figure/_p_i_egl-2_G574R_i_is_a_gain-of-function_allele_and_disrupts_EGF-induced_sleep_p_/34049539 CC BY 4.0