Skip to main content
. 2018 Nov 26;7:e43326. doi: 10.7554/eLife.43326

Figure 3. Inhibition of endocytosis in surface glia is sufficient to increase sleep.

(A) Total sleep at 18°C for female flies expressing Shibire (UAS-20xShi.ts1) by, from left to right, Alrm-GAL4 (n = 16, each genotype), MZ0709-GAL4 (n = 16–32), MZ0097-GAL4 (n = 16–32), NP2222-GAL4 (n = 16–32), NP6293-GAL4 (n = 29–32), moody-GAL4 (n = 16). MZ0709, MZ0097, and NP2222 experiments were loaded simultaneously and therefore share the same UAS-Control, graphed repeatedly for each to aid comparison. Significance markings shown for GAL4s in which the experimental group differed significantly from both controls. (B) Conditional expression of UAS-20xShi.ts1 using tubGal80.ts;Rab9-GAL4 at permissive (18°C) and restrictive temperature (30°C, red shading) (n = 9–19 females, mean of 4 days). (C) Exclusion of glial expression by Repo-Gal80 from the conditional expression of UAS-20xShi.ts1 by tubGal80.ts;Rab9-GAL4 at restrictive (18°C) and permissive temperature (30°C) (n = 15–16, mean of 4 days). One-way ANOVA with Holm-Sidak post-hoc test, *p<0.05, **p<0.01, *** is p<0.001. Error bars represent standard error of the mean (SEM).

Figure 3.

Figure 3—figure supplement 1. Glial adult-specific Shibire induction increases sleep and is reversible.

Figure 3—figure supplement 1.

(A) Day sleep at 18°C for female flies expressing Shibire (UAS-20xShi.ts1) by, from left to right, Alrm-GAL4 (n = 16, each genotype), MZ0709-GAL4 (n = 16–32), MZ0097-GAL4 (n = 16–32), NP2222-GAL4 (n = 16–32), NP6293-GAL4 (n = 29–32), moody-GAL4 (n = 16). MZ0709, MZ0097, and NP2222 experiments were loaded simultaneously and therefore share the same UAS-Control, graphed repeatedly for each to aid comparison. Significance markings shown for GAL4s in which the experimental group differed significantly from both controls. (B) Conditional expression of UAS-20xShi.ts1 using UAS-tubGal80.ts;Repo-GAL4 at restrictive (18°C), then permissive temperature (30°C, red shading), and back to restrictive (n = 14–16 females, mean of 3–4 days). One-way ANOVA with Holm-Sidak post-hoc test, *p<0.05, **p<0.01, *** is p<0.001. Error bars represent standard error of the mean (SEM).
Figure 3—figure supplement 2. Expression patterns of Rab9-G4 and the BBB drivers.

Figure 3—figure supplement 2.

(A) Superficial z-slices from confocal images of brains expressing UAS-nGFP driven by Rab9-GAL4, moody-GAL4 (subperineurial glia), and NP6293-GAL4 (perineurial glia) reveals the characteristically large and sparse nuclei of subperineurial glia. (B) BBB driver expression of UAS-CD8::GFP without and with Repo-Gal80 (revealing neuronal expression). Large images are single slices from the middle of the brain, insets are max projections of the entire brain. Laser power settings were maintained for comparison except in NP6293-GAL4 > UAS GFP+Repo-Gal80 images where they are higher to reveal weak expression which is otherwise not visible at all.
Figure 3—figure supplement 3. Perineurial glia are sufficient for Shibire rebound phenotype.

Figure 3—figure supplement 3.

Sleep during deprivation for NP6293-GAL4 and moody-GAL4 driving UAS-20xShi.ts1 at 18°C for female flies (n = 16–30). One-way ANOVA with Holm-Sidak post-hoc test, *p<0.05, **p<0.01, *** is p<0.001. Error bars represent standard error of the mean (SEM).