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. 2022 Jun 23;11:e74327. doi: 10.7554/eLife.74327

Figure 7. PPM3 convey enhancing homeostatic signal to R5 ellipsoid body neurons (A) Cartoon illustrating link between LNds and 5th sLNv and dFB via with PPM3 and R5 intermediates.

(B) GFP Expression pattern of split Gal4 line that labels Glu+ DN1ps (R58H05 AD; R48H04 DBD >GFP) at 20 x. (C–H) Comparison of sleep lost, baseline sleep, and sleep gain following deprivation at morning and evening timepoints modulating neurons linking LNd activity to the EB. Morning times are matched with evening time points with similar baselines. (C) Flies expressing an inactive form of tetanus toxin in PPM3 neurons (R92G05>TNTi)(N=45) exhibit greater rebound in the morning than at a matched evening time point (p<0.0001, paired t-test). (D) Silencing PPM3 neurons with an active form of tetanus toxin (R92G05>TNTa)(N=27) resulted in no significant difference between matched morning/evening time points (p>0.10, paired t-test). (E) Flies expressing an inactive form of tetanus toxin in R5 neurons (R58H05 AD; R48H04 DBD >TNTi) (N=21) exhibit greater rebound in the morning than at a matched evening time point (p<0.01, paired t-test). (F) Silencing R5 neurons with tetanus toxin (R58H05 AD; R48H04 DBD >TNTa) (N=16) resulted in no significant difference in sleep gain for matched morning and evening time points (p>0.70, paired t-test). (G) Flies expressing an inactive form of tetanus toxin in the dFB (R23E10>TNTi) (N=30) exhibit greater rebound in the morning than at a matched evening time point (p<0.0001, paired t-test). (H) Silencing dFB neurons with tetanus toxin (R23E10>TNTa) (N=12) resulted in no significant difference between morning and evening time points (p>0.45, paired t-test).

Figure 7—source data 1. PPM3 convey enhancing homeostatic signal to R5 ellipsoid body neurons.

Figure 7.

Figure 7—figure supplement 1. Silencing of PPM3, dFB and R5 reduces rebound in the morning and evening (A, B) Comparison of sleep gain at morning (A) and evening (B) timepoints between flies with silenced neurons (TNTa) and their controls (TNTi).

Figure 7—figure supplement 1.

Flies expressing an inactive form of tetanus toxin in PPM3 neurons (R92G05>TNTi)(N=45) exhibit greater rebound than flies with silenced PPM3 neurons (R92G05>TNTa)(N=27) in the morning (A) (p<0.0001, t-test) and evening (B) (p<0.0001, t-test). Flies expressing an inactive form of tetanus toxin in R5 neurons (R58H05 AD; R48H04 DBD >TNTi) (N=21) exhibit greater rebound than flies with R5 neurons silenced (R58H05 AD; R48H04 DBD >TNTa) (N=16) in both the morning (A) (p<0.0001, t-test) and evening (B) (p<0.0001, t-test). Flies expressing an inactive form of tetanus toxin in the dFB (R23E10>TNTi) (N=30) than flies with dFB neurons silenced (R23E10>TNTa)(N=12) in the morning (A) (p<0.0001, t-test) and evening (B) (p<0.0001, t-test). Data are means +/- SEM.
Figure 7—figure supplement 1—source data 1. Silencing of PPM3, dFB, and R5 reduces rebound in the morning and evening.
Figure 7—figure supplement 2. Standardized time points show similar effects to points with matched baseline sleep (A–C) FD sleep during two baseline time periods (sleep lost and baseline sleep) and sleep gain for rebound occurring at ZT1.5 and ZT 9.5.

Figure 7—figure supplement 2.

(A) Flies expressing an inactive form of tetanus toxin in PPM3 neurons (R92G05>TNTi)(N=45) exhibit greater baseline sleep during the rebound period at ZT9.5 (p<0.001 paired t-test) and exhibit significantly greater rebound at ZT 1.5 compared to ZT9.5 (p<0.0001, paired t-test).(B) Flies with PPM3 neurons silenced with the active form of tetanus toxin (R92G05>TNTa)(N=27) exhibit significantly less sleep lost (p<0.05, paired t-test) and sleep gain (p<0.0001, paired t-test) at ZT 9.5 compared to ZT1.5. Data are means +/- SEM.
Figure 7—figure supplement 2—source data 1. Standardized time points show similar effects to points with matched baseline sleep.