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. 2019 Feb 25;8:e42786. doi: 10.7554/eLife.42786

Figure 4. Sleep promoting vFB neurons activate DAN-aSP13.

(A) (left) Sleep profile of males (R23E10-GAL4 > UAS-TrpA1) upon activation of dFB neurons (red, n = 16) and control males with dFB neurons not activated (black, n = 16). Sleep time was plotted in 30 min bins. White and black areas indicate 12 hr light and dark periods, respectively. (right) Normalized calcium levels (dF/F) in DAN-aSP13 upon activation of dFB (R23E10-GAL4 > UAS-Chrimson88, R58E02-LexA > LexAop-GCamP6s). DAN-aSP13 activity in individual flies is shown in colored thin lines, and the mean trace is shown in thick magenta line with SEM indicated by shaded area. (B) (left) Sleep profile of males (VT03687-GAL4 > UAS-TrpA1) upon activation of vFB neurons (red, n = 16) and control males with vFB neurons not activated (black, n = 16). Sleep time was plotted in 30 min bins. White and black areas indicate 12 hr light and dark periods, respectively. (right) Normalized calcium levels (dF/F) in DAN-aSP13 upon activation of vFB (VT036875-GAL4 > UAS-Chrimson88, R58E02-LexA > LexAop-GCamP6s). DAN-aSP13 activity in individual flies is shown in colored thin lines, and the mean trace is shown in thick blue line with SEM indicated by shaded area. (C) (left) Sleep profile of males (VT036875-GAL4 > UAS-TrpA1, R58E02-GAL80) upon activation of vFB neurons (red, n = 16) and control males with vFB neurons not activated (black, n = 16). Sleep time was plotted in 30 min bins. White and black areas indicate 12 hr light and dark periods, respectively. (right) Normalized calcium levels (dF/F) in DAN-aSP13 upon activation of vFB (VT036875-GAL4 > UAS-Chrimson88, R58E02-GAL80, R58E02-LexA > LexAop-GCamP6s). DAN-aSP13 activity in individual flies is shown in colored thin lines, and the mean trace is shown in thick blue line with SEM indicated by shaded area. (D) (left) Sleep profile of (SS57264-GAL4 > UAS-TrpA1) males upon activation of vFB neurons (red, n = 16) and control males with vFB neurons not activated (black, n = 16). Sleep time was plotted in 30 min bins. White and black areas indicate 12 hr light and dark periods, respectively. (right) Normalized calcium levels (dF/F) in DAN-aSP13 upon activation of vFB (SS57264-GAL4 > UAS-Chrimson88, R58E02-LexA > LexAop-GCamP6s). DAN-aSP13 activity in individual flies is shown in colored thin lines, and the mean trace is shown in thick blue line with SEM indicated by shaded area. (A–D) (right panels) Red line indicates the time window of the light stimulus. P value represents the probability that the mean dF/F of pre-stimulation (10 s) and the mean dF/F during stimulation has the same median across flies (tested by Wilcoxon rank sum test, sample size indicated with n value).

Figure 4.

Figure 4—figure supplement 1. Specific class of vFB neurons acutely induces sleep.

Figure 4—figure supplement 1.

(A) Three single FB neural cell types were manually traced in MCFO data: dFB (magenta), vFB (blue) and mFB (grey). Confocal images registered to standard brain of four FB lines in Figure 4 and superimposed on the standard brain. (B) R23E10-GAL4 (C) VT036875-GAL4 (D) VT036875-GAL4, 58E02-GAL80 (E)SS57264 driving UAS-myrGFP or UAS-Chrimson88-tdTomato. (F) Total amount of sleep of the experienced and naïve males quantified per 30 min time period upon activation with CsChrimson (activated, red, n = 8–9 and not activated, black, n = 8–9). (G) Sleep profile of the genetic control males (pBDP-GAL4 > UAS-TrpA1) (30°C, red, n = 48) and (20°C, black, n = 48). Sleep time was plotted in 30 min bins. White and black areas indicate 12 hr light and dark periods, respectively.