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. 2021 Sep 17;10:e67670. doi: 10.7554/eLife.67670

Figure 8. AMsh phagocytic activity affects ASER and ASH sensory functions.

(A) Chemotactic indexes of four independent assays made with 30 N2 or AMsh::DYN-1(K46A) transgenics in linear 5 mM/cm NaCl gradients. The chemoattraction to NaCl was lost in transgenics expressing AMsh::DYN-1(K46A). One-way ANOVA, multiple comparisons corrected by Tukey test. (B) Chemotactic indexes of four independent assays made with 30 N2 or AMsh::DYN-1(K46A) transgenics in linear 5 mM/cm CuSO4 gradients. Animals expressing AMsh::DYN-1(K46A) did not show avoidance behavior to CuSO4. One-way ANOVA, multiple comparisons corrected by Tukey test. (C) Chemotactic indexes of three independent assays made with an average of >100 N2, che-2, or AMsh::DYN-1(K46A) transgenics in a gradient of the volatile attractant isoamyl alcohol (IAA). The top of the gradient was spotted with [10–2] IAA. AMsh::DYN-1(K46A) did not affect IAA chemotaxis. One-way ANOVA, multiple comparisons corrected by Dunnett’s test. (D) Thermotactic behavior in one temperature gradient assays made with >400 N2, ttx-3(ks5) or AMsh::DYN-1(K46A) transgenics. AMsh::DYN-1(K46A) did not consistently affect thermotaxis. (E) Transgenic animals expressing ChR2(H134R) in ASH sensory neurons (ASH::ChR2(H134R); lite-1) exhibit fast reversal (minimum 1–2 backward head swings) in response to blue light exposure (15 mw/mm2). This response was only observed when animals were raised in the presence of trans-retinal (TR+). Control groups were done using the same strain and same stimulation but were raised in the absence of trans-retinal (-TR). Expression of AMsh::DYN-1(K46A) does not modify this avoidance response. Kruskal–Wallis test, multiple comparison corrected by Dunn’s test.

Figure 8.

Figure 8—figure supplement 1. Experimental procedure followed for chemotaxis and thermotaxis assays.

Figure 8—figure supplement 1.

(A) NaCl chemotaxis gradient was generated by juxtaposition of two layers of agar solution, the bottom layer contained 50 mM NaCl and the top layer did not contain NaCl, effectively creating a 5 mM/cm linear gradient. (B) CuSO4 chemotaxis gradient was generated by juxtaposition of two layers of agar solution, the bottom layer contained 5 mM CuSO4 and the top layer did not contain CuSO4, effectively creating a 0.5 mM/cm linear gradient. (C) Scoring method used for both chemotaxis assays to soluble chemicals. Animal position was marked on the plate when the assay finished, then chemotaxis indexes (C.I.) were scored according to the formula shown above. (D) Scoring method used for chemotaxis to volatile odorant isoamyl alcohol (IAA). Animal position was marked on the plate when the assay finished, then chemotaxis indexes (C.I.) were scored according to the formula shown above. Spots A and B correspond to the test spots (IAA [10–2]) and spots C and D to the control spots (EtOH). Spot position was defined at 0.5 cm away from the plate edge. Animals were placed in the landing zone (gray dashed circle in the center), those that did not leave the landing zone were excluded from the scoring. (E) Thermotaxis assay was done in 9 cm plates divided into four sections each corresponding to an increasing temperature from left to right. Animals were initially positioned in the landing zone, indicated by a gray dashed rectangle in the center and allowed to navigate through the thermotactic gradient.