RNA-Seq of whole Drosophila larval central nervous systems (mid-3rd instar stage, central brain + ventral nerve cord) with panneuronal ISWI knockdown and controls. RNA-Seq of whole Drosophila larval central nervous systems (mid-3rd instar stage, central brain + ventral nerve cord) with panneuronal ISWI knockdown and controls

Sleep disruptions are among the most commonly-reported symptoms across neurodevelopmental disorders (NDDs), but mechanisms linking brain development to normal sleep are largely unknown. From a Drosophila screen of human NDD-associated risk genes, we identified the chromatin remodeler Imitation SWItch/SNF (ISWI) to be required for adult fly sleep. To better understand the mechanisms by which ISWI regulates sleep, we performed RNA-Seq analysis on mid-3rd instar larval central nervous systems (central brain + ventral nerve cord) in the setting of pan-neuronal ISWI knockdown (elav-GAL4>UAS-ISWI RNAi) compared to controls (elav-GAL4;+). We chose mid-3rd instar because temporal mapping revealed ISWI knockdown during this pre-adult developmental stage led to adult fly sleep deficits. Overall design: Experimental: elav-GAL4>UAS-ISWI RNAi (4 replicates) versus controls: elav-GAL4; + (3 replicates). All samples were from 4 days post-egg lay (the mid-3rd instar larval stage), with at least 40 CNS per replicate.

睡眠障碍是神经发育障碍（Neurodevelopmental Disorders, NDDs）中最常被报告的症状之一，但将大脑发育与正常睡眠联系起来的机制在很大程度上仍未被阐明。我们通过针对人类神经发育障碍相关风险基因的果蝇（Drosophila）筛选实验，鉴定出染色质重塑因子（chromatin remodeler）Imitation SWItch/SNF（ISWI）是成年果蝇维持正常睡眠所必需的基因。为深入解析ISWI调控睡眠的分子机制，我们对泛神经元ISWI敲低组（elav-GAL4>UAS-ISWI RNAi）与对照组（elav-GAL4;+）的3龄中期幼虫中枢神经系统（central nervous system, CNS，包含中枢脑与腹神经索）开展了RNA测序（RNA-Seq）分析。我们选择3龄中期作为样本采集时期，是因为时序图谱研究显示，在此成虫前发育阶段敲低ISWI会导致成年果蝇出现睡眠缺陷。本研究的整体实验设计如下：实验组为elav-GAL4>UAS-ISWI RNAi（4个生物学重复），对照组为elav-GAL4; +（3个生物学重复）。所有样本均采集自产卵后4天的3龄中期幼虫阶段，每个重复样本至少包含40个中枢神经系统组织。