Extended Data Fig. 8: Sleep deprivation affects synapse number in tectal neuron subtypes.

a, Percentage change of total sleep (left) and average sleep bout length (right) of each larva (dots) in the 6 hr post SD (ZT18-24, 7dpf), normalized to the circadian-matched time at 6 dpf. The black lines depict the average ± SEM. *P < 0.02, one-way ANOVA. b, The SD method did not alter circadian clock phase as measured by the bioluminescence driven by a Tg(per3-luc) reporter line for the clock gene per3 expression. The detrended per3 bioluminescence rhythms ( ± 95%CI) remained in phase for both SD (n = 14 larvae) and control (n = 12) larvae over multiple days of constant dark conditions. Circadian time (CT = 0 last lights ON transition). c, The percentage change in synapse number within each neuron between imaging sessions at ZT14 and ZT18, and between imaging at ZT18 and ZT24. d, Average (68%CI) for net synapse change per hour for FoxP2.A tectal subtypes in control or sleep deprived larvae. Type 3, but not Type 4 neurons significantly gain synapses after SD (Mixed ANOVA, post-hoc Benjamini-Hochberg, one tailed **p = 0.01, g = 1.266) and subsequently lose them (p = 0.014, g = −1.034) relative to controls. Type 2 lacks enough matched controls to assess. e, Sleep amount for early and late sleepers in the early (ZT14-18) and late (ZT18-24) phase of the night (5 independent experiments). The black lines depict the average ± SEM. f, For each neuron/larva, changes in synapse number during extended wakefulness did not correlate with either the subsequent total sleep or average sleep bout lengths (mean ± 95% CI). g, Changes in synapse numbers for each neuron/larva did not significantly correlate with the average sleep bout lengths during the early and late night of controls, or after SD (mean ± 95% CI). *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001, Mixed ANOVA interaction (condition*time), post-hoc Benjamini-Hochberg, two tailed.

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