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. 2019 Aug 30;19:1037–1047. doi: 10.1016/j.isci.2019.08.050

Figure 1.

Figure 1

csn7 Is an Allele of npr-1

(A) The protein sequence of the second predicted extracellular loop of NPR-1 (de Bono and Bargmann, 1998) is shown, aligned with human NPY receptors. csn7 harbors a Tyrosine substitution for the conserved Cys178. T-Coffee Multiple Sequence Alignment and BoxShade visualization tool.

(B) npr-1 mutants are defective in stress-induced sleep following exposure to Cry5B pore-forming toxin. Animals were exposed to Cry5B toxin for 15 min (shading) and examined for behavioral quiescence, defined as cessation of all movement and feeding. The majority of quiescence following Cry5B exposure is ALA dependent, as is evidenced by ALA-impaired ceh-17 mutants. npr-1(csn7) and the npr-1(lf) alleles ad609 and ky13 initially show SIS defects similar to ceh-17(lf) but over time engage in partially penetrant SIS. At least three trials of 25 young adult hermaphrodites were performed for each genotype. Mean and SEM are shown. **p < 0.01, ***p < 0.001, ****p < 0.0001 csn-7 versus wild-type; φ p < 0.0001 npr-1(ad609) versus ceh-17(np1), two-way repeated measures (RM) ANOVA with Sidak's multiple comparisons test. (C and D) npr-1(lf) arousal during SIS is robust to variations in baseline locomotor activity.

(C) Comparison of wild-type and npr-1(lf) locomotion during and outside of SIS. For Cry5B-SIS, animals were exposed to either E. coli OP50 (mock controls) or Cry5B-expressing E. coli for 15 min before examination. For UV-SIS in the absence of food, animals were rinsed and transferred to NGM plates lacking peptone and bacteria, exposed to UV for 1 min or mock treated, and examined 60 min later. Data points indicate individual animals. Mean and SEM are shown. ***p < 0.001, ****p < 0.0001, one-way ANOVA with Sidak's multiple comparisons test.

(D) Striking differences in SIS are seen between wild-type and npr-1(lf) animals under conditions with large (on food) or small (off food) differences in baseline locomotion. Each data point represents the fraction of quiescent animals (no feeding, no locomotion) in one trial of 25 young adult hermaphrodites. Mean and SEM of multiple independent trials are indicated. ****p < 0.0001 versus wild-type, Student's t test.