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. 2018 Oct 17;9:4311. doi: 10.1038/s41467-018-06819-5

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© The Author(s) 2018

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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Fig. 4 — GLR-1 and GLR-5 in AIB are required for quinine-induced reversal initiation and pumping inhibition, respectively. a glr-1 is required for quinine-induced reversal initiation, which can be rescued by transgenic expression of wild-type glr-1 gene in AIB. Error bars: s.e.m. n = 6 groups, 10 worms/group. ***p < 0.001 (t test). b glr-5 is required for quinine-induced pumping inhibition, which can be rescued by transgenic expression of wild-type glr-5 gene in AIB. Error bars: s.e.m. n = 16 worms. ***p < 0.001 (t test or Wilcoxon test). c Optogenetic stimulation of ASH by blue light (470 nm; 0.03 mW mm⁻²; 30 s pulse) triggered reversals in wild-type worms, but not in glr-1 mutant worms. The transgenic animals cultured on ATR-free plates were taken as control. d Optogenetic stimulation of ASH by blue light inhibited pumping rate in wild-type worms, but not in glr-5 mutant worms. The transgenic animals cultured on ATR-free plates were taken as control. c, d Error bars: s.e.m.. Reversal index, n ≥ 3 groups, 10 worms/group; Pumping rate, n ≥ 5 worms. ***p < 0.001 (t- test). e, g AIB calcium dynamics induced by low (e) and high concentration of quinine (g), respectively, in freely behaving worms acquired by the iCaN system. Traces are ordered according to the time of quinine application (dash lines). e, f AIB calcium transients induced by 1 mM quinine were largely abolished in glr-1 mutant worms. e Heat maps. f Representative traces (upper) and peak calcium changes (lower). Error bars: s.e.m.. n ≥ 16. ***p < 0.001 (t- test). N.S. represents no significant difference. g, h AIB calcium transients induced by 5 mM quinine in glr-1 and glr-5 mutant worms. g Heat maps. h Representative traces (upper), peak calcium changes and FHWMs for AIB calcium dynamics (lower). Notice that the peak amplitudes of AIB [Ca²⁺]_i elevations in neither glr-1 nor glr-5 mutant worms shown significant difference from wild-type worms, but the full width at half maximum (FWHM) were much longer in glr-1 mutant worms than that in glr-5. Error bars: s.e.m. n ≥ 17. **p < 0.01. ***p < 0.001 (Wilcoxon test)