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. 2018 Mar 12;7:e26016. doi: 10.7554/eLife.26016

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© 2018, Burgos et al

This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.

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Figure 1. — (A) Schematic showing the Drosophila larval CNS. Red scaffold represents class IV (cIV) projections. Enlarged transverse section through ventral nerve cord (VNC) is shown below. Color-coded regions depict modality specific locations where sensory axons terminate and motor neuropil. (B) 412-Gal4 drives expression in interneurons in the VNC (arrowhead) and neurons in the brain lobes (arrow), anti-dsRed, green. (C–C') Dorsal view of the morphology of DnB neuron in segment A3. Medial process is indicated by an arrow and lateral projection by an arrowhead. An asterisk marks the cell body. (C') Transverse section of neuron in C. (D–F) Intersectional strategy to target GFP either to D) the brain and VNC, (E) brain only or F) VNC only. Green channel shows anti-GFP labeling. (G) Percent exhibiting nocifensive rolling during dTrpA1 activation of subsets of 412-Gal4 neurons corresponding to panels D-F. (H) Schematic of cIV nociceptors (left) and location of 412-Gal4 VNC neurons (right). (I) Schematic of different motor behaviors observed in response to cIV or 412-Gal4 VNC activation. Body bend only, B, larvae entered a curved C-shape but did not roll or crawl; Rolling, R, animals entered C-shape and performed 360˚ rotations; Bend + crawling, BC, larvae attempted to crawl while remaining in a C-shape. Red arrows show direction of locomotion. (J) Total amount of time spent in bent-body positions (B + R + BC) upon dTrpA1-induced activation of cIV neurons and 412-Gal4 VNC neurons. (K) Percent of time upon dTrpA1 activation spent in bent-body positions with crawling (black) rolling (blue) or paused (bend-only, gray). (L) Percent of time spent during 29 s trial in bent-body positions: bend-crawl, rolling, or bend-only. (M) Plot showing length of bend-crawl bouts in seconds upon cIV or 412-Gal4 VNC activation. Box plots show median (middle line) and 25th to 75th percentiles with whiskers representing 10 to 90 percentiles. P values are indicated as *p<0.05, ***p<0.001, as determined by One-way ANOVA with Tukey’s multiple comparison’s test (J), Kruskal-Wallis with Dunn’s correction for multiple testing (L), or Mann-Whitney (M). Scale bars = 50 µm (B, D–F), 20 µm (C). (See also Figure 1—figure supplements 1–4).

Figure 1—source data 1. Summary table of graph data and statistical testing for thermogenetic activation experiments.
Genotypes, number of animals tested, graph data and statistical testing presented for thermogenetic activation experiments.

elife-26016-fig1-data1.xlsx^{(33.6KB, xlsx)}

DOI: 10.7554/eLife.26016.007

Figure 1—figure supplement 1. — (A) Schematic showing the Drosophila larval CNS. Red scaffold represents class IV (cIV) projections. Enlarged transverse section through ventral nerve cord (VNC) is shown below. Color-coded regions depict modality specific locations where sensory axons terminate and motor neuropil. (B) 412-Gal4 drives expression in interneurons in the VNC (arrowhead) and neurons in the brain lobes (arrow), anti-dsRed, green. (C–C') Dorsal view of the morphology of DnB neuron in segment A3. Medial process is indicated by an arrow and lateral projection by an arrowhead. An asterisk marks the cell body. (C') Transverse section of neuron in C. (D–F) Intersectional strategy to target GFP either to D) the brain and VNC, (E) brain only or F) VNC only. Green channel shows anti-GFP labeling. (G) Percent exhibiting nocifensive rolling during dTrpA1 activation of subsets of 412-Gal4 neurons corresponding to panels D-F. (H) Schematic of cIV nociceptors (left) and location of 412-Gal4 VNC neurons (right). (I) Schematic of different motor behaviors observed in response to cIV or 412-Gal4 VNC activation. Body bend only, B, larvae entered a curved C-shape but did not roll or crawl; Rolling, R, animals entered C-shape and performed 360˚ rotations; Bend + crawling, BC, larvae attempted to crawl while remaining in a C-shape. Red arrows show direction of locomotion. (J) Total amount of time spent in bent-body positions (B + R + BC) upon dTrpA1-induced activation of cIV neurons and 412-Gal4 VNC neurons. (K) Percent of time upon dTrpA1 activation spent in bent-body positions with crawling (black) rolling (blue) or paused (bend-only, gray). (L) Percent of time spent during 29 s trial in bent-body positions: bend-crawl, rolling, or bend-only. (M) Plot showing length of bend-crawl bouts in seconds upon cIV or 412-Gal4 VNC activation. Box plots show median (middle line) and 25th to 75th percentiles with whiskers representing 10 to 90 percentiles. P values are indicated as *p<0.05, ***p<0.001, as determined by One-way ANOVA with Tukey’s multiple comparison’s test (J), Kruskal-Wallis with Dunn’s correction for multiple testing (L), or Mann-Whitney (M). Scale bars = 50 µm (B, D–F), 20 µm (C). (See also Figure 1—figure supplements 1–4).

Figure 1—source data 1. Summary table of graph data and statistical testing for thermogenetic activation experiments.
Genotypes, number of animals tested, graph data and statistical testing presented for thermogenetic activation experiments.

elife-26016-fig1-data1.xlsx^{(33.6KB, xlsx)}

DOI: 10.7554/eLife.26016.007