Extended Data Fig. 3: Amino acid deprivation has no effect on CNMa expression in the fat body or the brain, whereas EAA deprivation but not NEAA deprivation influences CNMa expression in the gut.
From: Response of the microbiome–gut–brain axis in Drosophila to amino acid deficit
a, b, Representative confocal images (left), the relative signal intensity of native GFP fluorescence (middle) and the numbers of GFP-positive cells (right) in the fat body (a) and the brain (b; insets show the dorsal region) in flies carrying CNMa-Gal4 and UAS-GFP (green) that were fed a high-amino-acid versus a low-amino-acid diet. Nuclei in the fat body counterstained with DAPI (a, left), and the brain stained with anti-GFP antibody (green) and the neuropil marker nc82 (magenta) (b, left) are shown. The numbers of GFP-positive cells are as follows: 41.00 ± 7.28 (high amino acids) and 41.54 ± 8.16 (low amino acids) in the fat body; 3.96 ± 1.48 (high amino acids) and 3.29 ± 1.33 (low amino acids) in the brain. Scale bars, 50 μm (a, b); 10 μm (b, inset). c, d, EAA deprivation, but not NEAA deprivation, induced CNMa expression (c) and possibly mobilized intracellular calcium (d). Quantifications of GFP fluorescence in the R2 region of the anterior midgut in CNMa-GAL4>UAS-GFP flies (c) or CNMa-GAL4>CaLexA flies (d) that were fed with an EAA-deficient versus a NEAA-deficient diet for two days. e, f, Deficiency of a single EAA in a diet induced CNMa expression (e) and possibly mobilized intracellular calcium (f). Quantifications of GFP fluorescence in the R2 region in CNMa-GAL4>UAS-GFP flies (e) or CNMa-GAL4>CaLexA flies (f) that were fed with a holidic diet lacking a single EAA for three days. Data are mean ± s.e.m. P values are indicated; unpaired two-tailed t-test in a, b; one-way ANOVA with Tukey’s post-hoc test in c–f. Sample sizes and statistical analyses are shown in Supplementary Table 1.
