Figure 3.

Astrocytic aay is a novel regulator of water consumption

(A) Schematic for temperature control of GAL80^ts/GAL4 driven expression of UAS-RNAi or UAS-cDNA transgenes with CAFE test. Orange section of line indicates period of water restriction.

(B) Water consumption in the CAFE assay of flies with RNAi knock down of candidate genes. Dotted blue line indicates normalization to control Repo-GAL4 flies, equal to 1. ^∗p < 0.05, ^∗∗p < 0.01 two-tailed Mann-Whitney test n_RNAi = 20–25, n_Repo-GAL4 = 50.

(C) Water consumption in CAFE for UAS overexpression of targets. ^∗∗p < 0.01 two-tailed Mann-Whitney test n_RNAi = 17–29, n_Repo-GAL4 = 43.

(D) Schematic for temperature control of GAL80^ts/GAL4-driven UAS-RNAi or UAS-cDNA transgenes with manual water feeding assay. Orange section of line indicates period of water restriction.

(E) RNAi knockdown of aay reduces water consumption (n = 14–16).

(F) Overexpression of aay increases water consumption (n = 11–13).

(G) Astrocyte-specific RNAi knockdown of aay reduces water consumption (n = 13 and 14).

(H) RNAi knockdown of aay in perineurial glia does not alter water consumption (n = 18–20).

(I) Preventing vesicular transmission with tetanus-toxin (TNT) expression in astrocytes reduces water consumption (n = 12–19).

(J) Temperature regimen for TrpA1 activation of astrocytes in (K) and (L). Orange section of line indicates period of water restriction.

(K) Astrocyte activation for 40 min increases water consumption (n = 16–22). ^∗∗p < 0.01, Kruskal-Wallis ANOVA with Dunn’s multiple comparisons test.

(L) Astrocyte activation for 10 min increases water consumption (n = 28–30). ^∗∗p < 0.01, Kruskal-Wallis ANOVA with Dunn’s multiple comparisons test. Normality was assessed using Shapiro-Wilk test. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001 Ordinary one-way ANOVA with Dunnett’s multiple comparisons test, unless otherwise stated. Individual data points are single flies. Data are mean ± standard error of the mean (SEM).

See also Figure S3.