Fig. 4: SELKs promote feeding via acetylcholine.
From: Feeding decision-making by a single neuron via disparate neurotransmitters
a The effects of SELK inhibition on feeding. A flyPAD feeding assay with flies in which GtACR1 expression is driven by Lk-Gal4 (dark red), SELK1 (red) or SELK2 (light red). Genetic controls bearing either allele alone, or controls bearing both alleles but not exposed to light, are shown in shades of gray. The light is kept on throughout the experiment for continuous optogenetic inhibition. One-way ANOVA. Adjusted P values are 0.0004 for Lk > GtAcR1 vs. UAS-GtAcR1, 0.038 for Lk > GtAcR1 vs. Lk-Gal4, <0.0001 for Lk > GtAcR1 vs. Lk-Gal4 > GtAcR1 no light, <0.0001 for SELK1 > GtACR vs. UAS-GtAcR1, 0.036 for SELK1 > GtACR vs. SELK1, <0.0001 for SELK1 > GtACR vs. SELK1 > GtAcR1 no light, <0.0001 for SELK2 > GtAcR1 vs. UAS-GtAcR1, 0.008 for SELK2 > GtAcR1 vs. SELK2, and <0.0001 for SELK2 > GtAcR1 vs. SELK2 > GtAcR1 no light. b SELKs revealed by staining with an antibody against LK (magenta) are colabeled with an antibody against ChAT (green). Arrowheads mark the SELKs among the ChAT+ neurons. Scale bar: 10 μm. c In a flyPAD feeding assay with flies in which ChAT expression is knocked down in LK neurons (red), the total numbers of sips are plotted. Genetic controls are indicated in shades of gray. One-way ANOVA. Adjusted P values are 0.004 and 0.0007, respectively. d–f Quantification of PER upon light stimulation in optogenetic experiments. d Driving CsChrimson expression in sweet GRNs (magenta) and in neurons labeled by Lk-Gal4 (dark red), SELK1 (red) and SELK2 (light red). One-way ANOVA. Adjusted P values are <0.0001 for UAS-Chrimson vs. Gr64f > Chrimson, 0.008 for UAS-Chrimson vs. SELK1 > Chrimson, 0.020 for UAS-Chrimson vs. SELK2 > Chrimson, and <0.0001 for UAS-Chrimson vs. Lk > Chrimson. e Driving CsChrimson expression in Lk neurons via Lk-LexA either in the w1118 background (Lk+/+) (dark red) or in the LkCC9 mutant background (light red). Unpaired t-test with Welch’s correction, the two-tailed P value is 0.186. f Optogenetic activation of Lk neurons while the expression of ChAT is downregulated. Unpaired t-test with Welch’s correction. The two-tailed P value is 0.3388. Error bars indicate the standard error of the mean. In each panel of the behavioral experiments, the schematic presents the experimental paradigm, while the alleles expressed by each group of flies and the numbers of animals in each group are indicated in the tables below the plots. Boxplots indicate median (middle line), 25th and 75th percentile (box), bars represent maximum and minimum. For all panels, n indicates biological replicates. Source data are provided as a Source data file.
