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2026-10-01T17:47:02Z
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<p><i>egl-2</i> functions in the ALA neuron to regulate sleep.</p>
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
<p>(<b>A</b>) Quantification of sleep behavior as the fraction of quiescent worms for the indicated genotypes and conditions. Tissue- and cell-specific rescue experiments in which wild-type <i>egl-2</i> cDNA was expressed specifically in neurons (<i>Prgef-1</i>), in muscles (<i>Pmlc-2</i>), in the ALA neuron (<i>Pnlp-67</i>), or in the RIS neuron (<i>Pflp-11</i>), respectively. 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. Each dot represents one independent assay containing 15–25 animals on a single plate. Mean ± SEM are shown. ns, not significant; ****, p < 0.0001. One-way ANOVA with Dunnett’s multiple-comparisons test for post-heat shock samples, comparing each genotype to wild type. (<b>B</b>) Representative images of the head region of L4 animals of the indicated genotypes showing EGL-2 expression in the ALA neuron using a split-GFP strategy. Top: <i>ALAp::gfp1–10</i> alone; middle: <i>egl-2::7x gfp11</i> alone; bottom: <i>ALAp::gfp1–10; egl-2::7x gfp11</i>. White dashed ovals outline the ALA neuron cell body. A, anterior; D, dorsal. Scale bar, 20 μm. (<b>C-D</b>) Quantification of endogenous stress-induced sleep (SIS) and ALA neuron activity in animals for the indicated genotypes and conditions. Pre and post refer to before and 15 min after heat shock, respectively. All strains contain the GCaMP6s transgene for calcium imaging. (C) SIS is quantified as the fraction of quiescent worms of the indicated genotypes and conditions. Each dot represents one independent assay containing 15–25 animals on a single plate. Mean ± SEM are shown. ns, not significant; ****, p < 0.0001. One-way ANOVA with Dunnett’s multiple-comparisons test for post-heat shock samples, comparing each genotype with wild type. (D) ALA neuron activity is quantified as the ratio of GCaMP6s to mCherry fluorescence measured in the cell body (F<sub>green</sub>/F<sub>red</sub>). Sample sizes n = 28, 29, 29, 30, 28, and 26, from left to right. The dotted line indicates the threshold (0.76) for high ALA activity, defined as the wild-type pre-heat shock mean + 3x standard deviation (SD), corresponding to the upper 99.7% bound of baseline activity. ns, not significant; *, p < 0.05; ****, p < 0.0001. Two-way ANOVA with Dunnett’s multiple-comparisons test for comparing each genotype with wild type under the same condition. (<b>E</b>) Fraction of animals with high or low ALA activity at 15 min post-heat shock. The same post-heat shock samples analyzed in (D) were categorized as having high or low activity based on the threshold defined in (D). Numbers above bars indicate the total animals analyzed. ns, not significant; ****, p < 0.0001. Two-sided Fisher's exact test with Bonferroni correction for three pairwise comparisons.</p>
2026-10-01T17:46:48Z
Image
Figure
10.1371/journal.pgen.1012255.g003
https://figshare.com/articles/figure/_p_i_egl-2_i_functions_in_the_ALA_neuron_to_regulate_sleep_p_/34049530
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