Extended Data Fig. 9: Examples of manipulated single FoxP2.A:FingR(PSD95)+ neurons and clonidine and evidence that daytime drug treatment reduced sleep the following night.
From: Sleep pressure modulates single-neuron synapse number in zebrafish
a, left Example FoxP2.A:FingR(PSD95)+ tectal neurons imaged before (ZT14), immediately after (ZT18), and 6 h after (ZT24) sleep deprivation and control. Nuclei and synapses are FingR(PSD95)-GFP+ (green), and cellular morphology is labelled by mKate2f (magenta). Right, Higher magnification (dotted white box) showing the same dendritic segments at each time point, with examples of synapses lost (yellow arrows and dotted circles) or gained (blue arrows and circles). Note that, for illustrative purposes, the dendrites are depicted at a different angle in these higher magnification images. b, An example neuron before (ZT5) or after (ZT10) exposure to clonidine and 2-chloroadenosine. Scale bars: 15 μm (a, b left) and 5 μm (a, b right). c, Larvae (n = 80) exposed to lights OFF at mid-day (ZT8, first arrow in schematic) took longer to sleep (mean ± SEM) compared to lights OFF at the end of day (ZT14, 2nd arrow). ****P = 2.27 × 10−15, Kruskal-Wallis. d, Average locomotor activity ( ± 95%CI) on a 14 hr:10 hr LD cycle before, during, and after a 5 hr midday (ZT5-10, 7 dpf, shaded purple panel) exposure to melatonin (n = 31 larvae), clonidine (n = 32), or DMSO (n = 32). Data from two independent experiments. e, Larvae treated with either melatonin or clonidine from ZT5-10 had reduced and delayed sleep ( ± SEM) in first hour of the night (ZT14-15) compared to controls. *P < 0.05, **P < 0.01, ****P < 0.0001 Dunnett’s Test.
