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. 2020 Oct 19;10:17614. doi: 10.1038/s41598-020-74448-4

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© The Author(s) 2020

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Optogenetic activation of muscles (G7-GAL4, (a)–(c)), several sensory neurons (5-40-GAL4, (d)–(f)), and class IV multidendritic neurons (ppk-GAL4, (g)–(i)) by expression of CsChrimson. (a) Expression of CsChrimson::Venus by G7-GAL4 as observed by antibody staining (green). Magenta represents the motor neuron stained with anti-HRP (horseradish peroxidase), a neuronal membrane marker. Scale bar: 10 µm. (b)–(c) Stimulation of muscles in larvae expressing G7 > CsChrimson. (b) Representative images from video recordings before (left) and after (right) illumination with white light. (c) Normalised larval length for a sequence of 20 s stimulation for each colour and 60 s black period. (d) 5-40-GAL4 drives expression of Chrimson::Venus in the peripheral nervous system consisting of chordotonal (cho) neurons, external sensory (es) neurons, and multidendritic (md) neurons. Scale bar: 50 µm. (e)–(f) Stimulation of all sensory neurons in larvae expressing 5-40 > CsChrimson. Stimulation period: 20 s; black period: 30 s. (g) Using ppk-GAL4, Chrimson::Venus is exclusively expressed in class IV dendritic arborisation (C4da) neurons. Scale bar: 10 µm. (h)–(i) Stimulation of larvae expressing ppk > CsChrimson. (h) Image sequence before (t = 0 s) and after stimulation with white light displaying a characteristic rolling behaviour. (i) Percentage of larvae that were rolling over time. Stimulation period: 20 s; black period: 40 s. N: number of larvae; whiskers: s.e.m.; diamonds: mean. Significance calculated via one-sample two-tailed t-test: ns: not significant (p > 0.05), **p < 0.01, ***p < 0.001.