Fig. 1: Mechanical stimulation evokes ANO1-dependent calcium increases in CEM neurons.

a Micrograph showing four CEM neurons in the head of an adult male worm. Scale bars, 10 µm. b Nose touch evoked a calcium increase in CEM. Left: representative time-lapse images of GCaMP5.0- based calcium responses in CEM induced by nose touch; Right: calcium response indicated by fluorescence changes of GCaMP5.0. The arrows indicate the application of mechanical force (15 μm displacement). c The touch-evoked calcium responses in CEM were dependent on chloride ions and were abolished by NFA. Left: calcium responses; Right: maximum ΔF/F₀ changes. Solid lines show the average fluorescence changes and the shading indicates SEM. P values were calculated using the Brown-Forsythe and Welch ANOVA tests. d Chloride transporter mutants affected the nose touch-evoked calcium increases in CEM. Left: maximum ΔF/F₀ changes; Right: calcium responses. P values were calculated using the Brown-Forsythe and Welch ANOVA tests. e, f The absence of ANOH-1 or BEST-2 significantly reduced the mechanically activated chloride channels in CEM. Calcium responses (f); Maximum ΔF/F₀ changes (e). P values were calculated using the Kruskal-Wallis test. g Transgenic expression of either nematode or human anoctamin-1 genes rescued the nose touch-evoked calcium increases in CEM of anoh-1(tm4762) mutant male worms. Left: calcium responses; Right: maximum ΔF/F₀ changes. The mutations of anoh-1(K588A) and ANO1(K645A) will be further elucidated in the subsequent sections. P values were calculated using the Kruskal-Wallis test. Day 2 adult male animals were used in these experiments. The numbers of independent assays are indicated in each column of the panel. Each dot represents 1 animal. Data are presented as mean ± SEM. ns not significant, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. Source data are provided as a Source Data file.