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. 2017 Dec 13;7:17514. doi: 10.1038/s41598-017-17819-8

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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CSQ-1 deletion and the point mutation P319S affect pharyngeal pumping. (a) Expression of pcsq-1::csq-1::CFP in BWM cells and (b) pharynx muscle of the deletion mutant csq-1(ok2672). Scale bars: 50 μm and 20 μm, as indicated. Left panel: Fluorescence micrograph, right panel: DIC image; dashed lines indicate pharynx position. Structural features of the pharynx are labeled: TB (terminal bulb), I (isthmus), C (corpus), X (grinder). (c) Swimming cycles/min and (d) pump rate on food, of csq-1(ok2672) deletion mutants, as well as full length pcsq-1::csq-1::CFP rescue in pmyo-2::ChR2::mCherry background. Also analyzed (d) is the point mutation CSQ-1(P319S), compared to wt (number of animals tested is indicated at the base of each bar). (e) Number of pumps evoked at 3.7 Hz pulse frequency was counted for 100 light stimuli and averaged in 10 stimuli bins for the indicated number of animals. Original kymographs are depicted for wt (black), csq-1(ok2672) (red) and csq-1(gk876502) (blue; P319S). (f) Percentage of light stimuli inducing a pump across 100 light stimuli (data in e). (g) Percentage of pumps that were induced with a deviation of > 50 ms following a light stimulus (here defined as ‘jitter’). Shown (f and g) are wt (white), csq-1(ok2672) deletion mutant (grey) and P319S mutant (black). (h) Pump rate distribution (%) at 3.7 Hz pacing (white: > 4.5 Hz, blue: 3.3–4.5 Hz, green: 2.5–3.3 Hz, red: 1.6–2.5 Hz, grey: 0.5–1.6 Hz, black: 0 Hz) in the indicated csq-1 mutants, compared to wild type (wt) (n = 11–15). (i) Original EPG recordings of cut-head preparations and (j) mean of maximal pump rate of deletion and P319S mutant achieved in a stress test (470 nm, 10 ms pulses at stepwise increasing pulse rate (1 Hz steps, 5 s each step, 1–7 Hz), as indicated by blue tick marks) compared to wt (n = 10–23). Statistically significant differences, 1-way ANOVA and Bonferroni post-hoc test: ***p < 0.001; **p < 0.01; *p < 0.05.