Extended Data Fig. 10: CNMa expression and l-EAA preference are regulated by the microbiome and Tor signalling in the gut.
From: Response of the microbiome–gut–brain axis in Drosophila to amino acid deficit
a, Quantifications of GFP fluorescence in the R2 region of the gut of GF flies carrying CNMa-Gal4>UAS-GFP that had been mono-associated with Acetobacter that has a mutation in the proC gene (AcetoΔproC) or the leuB gene (AcetoΔleuB). Complementation of the AcetoΔleuB strain was achieved by re-introducing the leuB gene to generate AcetoΔleuB_leuB. Dietary complementation of AcetoΔleuB was achieved by adding 10 mM leucine to the diet. b, c, Representative confocal images (left) and quantifications of GFP fluorescence (right) in the R2 region of the gut (b) and two-choice preferences (c) of GF flies that had been mono-associated with Acetobacter that has a mutation in ilvA (AcetoΔilvA). Dietary complementation of AcetoΔilvA was achieved by adding 10 mM isoleucine to the diet. GF flies carrying CNMa-Gal4>UAS-GFP and GF w1118 flies were used in b and c, respectively. d, Quantifications of GFP fluorescence in the R2 region of the gut of GF flies carrying CNMa-Gal4>UAS-GFP that had been mono-associated with an engineered strain of L. plantarum WJL that is capable of producing BCAAs (LactoBCAA). Dietary complementation of L.plantarum WJL (LactoWT) was achieved by adding 10 mM of leucine, isoleucine and valine to the diet (LactoWT + BCAA). e, Representative confocal images (left) and quantifications of the GFP signal (right) in the anterior R2 midgut region of AcetoΔleuB mono-associated GF flies carrying CNMa-Gal4 and UAS-GFP (GFcontrol), and flies carrying CNMa-Gal4, UAS-GFP and UAS-TorWT (GFTor-WT) on 10% dietary yeast, measured by the native GFP fluorescence. f, Two-choice preferences of AcetoΔleuB mono-associated flies carrying enterocyte-specific NP1-Gal4 (control), and flies carrying NP1-Gal4 and UAS-TorWT (n = 20). Nuclei were stained with DAPI. Scale bars, 50 μm (b, e). Data are mean ± s.e.m. P values are indicated; one-way ANOVA with Tukey’s post-hoc test in a–d; unpaired two-tailed t-test in e, f. Sample sizes and statistical analyses are shown in Supplementary Table 1.
