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. 2018 Aug 2;7:e38554. doi: 10.7554/eLife.38554

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© 2018, Carreira-Rosario 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–C) Split1-Split3 lines driving expression of membrane localized Venus in the third instar CNS. Corazonin (not shown) labels a single neuron in segments T2-A6, and was used to identify VNC segment identity. The only neurons potentially common to all three lines are a pair of bilateral ventral, anterior, medial neurons (arrowheads). Maximum intensity projection of entire CNS shown. Anterior, up; scale bars, 50 μm. Genotypes: R49F02-Gal4^AD R53F07-Gal4^DBD UAS-Chrimson:mVenus (Split1); R49F02-Gal4^AD R53F07-Gal4^DBD tsh-lexA lexAop-killer zipper UAS-Chrimson:mVenus (Split2); ss01613-Gal4 UAS-Chrimson:mVenus (Split3). (D) Split1 activation induces backward locomotion. Genotype: R49F02-Gal4^AD R53F07-Gal4^DBD UAS-Chrimson:mVenus. (E) Split1, Split2, or Split3 activation induces backward locomotion. Number of backward or forward waves in third instar larvae over 15 s with or without Chrimson activation. N = 10 for all genotypes. Genotypes: pBD-Gal4 UAS-Chrimson:mVenus (Empty) and see A-C above for Split1-3 genotypes. (F) Split1, Split2, or Split3 activation induces backward locomotion. Percentage of time performing forward locomotion (green), backward locomotion (magenta) or paused (grey) in third instar larvae over 15 s with or without Chrimson activation. N = 5 for all genotypes. Genotypes, see A-C. (G) Split2 or Split3 silencing reduces initiation of backward locomotion. Backward waves induced by a noxious head poke, with or without active GtACR1. Genotypes: pBD-Gal4 UAS-GtACR1:mVenus (first two bars, n = 20), R49F02-Gal4^AD R53F07-Gal4^DBD tsh-lexA lexAop-killer zipper UAS-GtACR1:mVenus (middle two bars, n = 8), ss01613-Gal4 UAS-GtACR1:mVenus (last two bars, n = 25). (H) Split2 or Split3 neuron silencing stops ongoing backward locomotion. After each larva initiated a backward run (two backward waves), light was used to activate GtACR1 or a no Gal4 control, and the number of backward waves was counted. n = 6 for both groups; each bar represents the average of two trials for the same larva. See G for genotypes.