A genomic characterization of the response to sleep loss

睡眠门，或者说是对睡眠需求的解决。睡眠需求的解决反映了睡眠功能 对觉醒的反应(睡眠稳态)。关于睡眠/觉醒状态的控制，现在已经知道很多 表达和持续时间，但睡眠功能还不是很清楚。 基因的显著差异表达(DEG‘s；6,000/13000在额叶皮质的大块组织中表达；FC) 作为对6小时睡眠剥夺(SD)的反应，已经观察到。对中枢神经系统组织具有特殊意义， 睡眠调节基因本体支持对神经元兴奋性和传递的睡眠控制。 睡眠对细胞功能的调节可能会因细胞类型的不同而有所不同 然而，人们对睡眠障碍的细胞类型特异性及其对生物学内和生物学的影响了解甚少。 负责SD反应的细胞间通路。 ADO可能被认为是一种细胞间(神经胶质细胞到神经元)的中介，因为它的受体被激活， ADORA1是睡眠稳态慢波活动(SWA)所必需的，但它在睡眠相关基因表达中的作用 还没有经过检查。 提出了以下具体目标来描述关于睡眠缺失的基因组反应 DEGS的细胞类型特异性及其在细胞间和细胞内决定SD反应的途径 细胞内水平，在小鼠Fc。 1.比较睡眠缺失对细胞类型特异性mRNA表达的影响 以随机睡眠状态和恢复睡眠状态来表征细胞类型、特异性和共享性 C57BL/6J小鼠FC组织对睡眠缺失和恢复的基因表达反应 2.睡眠剥夺和睡眠丧失恢复诱导的细胞类型特异性开放染色质的鉴定 使用ATACseq来确定对睡眠缺失反应重要的潜在增强子和启动子位置。 3.ADORA1受体的腺苷激活在细胞类型对睡眠缺失的特异性基因组反应中的作用将 使用对ADO和神经胶质条件ADK无反应的条件性Adora1基因敲除进行检查 在清醒和睡眠时，脑部Ado和SWA的慢性升高会被击倒。 第一个目标可以提供关于细胞类型的睡眠功能的细胞靶点的基本见解 失眠反应的特异性和差异基因组本体论。第二个目标是确定 DNA基因座作为潜在的、细胞类型特定的、增强子和/或启动子位点。这些地点的标识是 对于SD激活的基因组通路的表征以及进一步的，是迈向 了解并优先考虑在非编码DNA区域中识别的基因座，以供将来验证。第三 AIM将测试Ado作为调节因睡眠不足而观察到的基因组变化的作用，以及 或者，它对稳态睡眠的调节作用可以从睡眠丧失转录组中分离出来。

Sleep gates, or is permissive to, the resolution of sleep need. The resolution of sleep need reflects sleep function induced in response to waking (sleep homeostasis). Much is now known about the control of sleep/wake state expression and duration, but sleep function is less well understood. Significant differential expression of genes (DEG’s; 6000/13000 expressed in bulk tissue from frontal cortex; FC) in response to 6 hours of sleep deprivation (SD) has been observed. Of particular significance for CNS tissue, the sleep modulated gene ontology supports sleep control of neuronal excitability and transmission. Sleep dependent modulation of cellular function is likely to vary depending on different cell-type specific roles yet, little is understood about the cell-type specificity of sleep DEGs and their implications for biological intra- and inter-cellular pathways responsible for the SD response. Ado may be implicated as an inter- cellular (glia to neuron) mediator of sleep need since activation of its receptor, ADORA1 is required for sleep-homeostatic slow wave activity (SWA) but its role in sleep-related gene expression has not been examined. The following specific aims are proposed to characterize the genomic responses to sleep loss with respect to the cell-type specificity of DEGs and the pathways determining their SD response at an intercellular and intracellular level, in the mouse FC. 1.The differential expression of cell-type specific mRNA expression in response to sleep-loss will be compared to ad-libitum sleep condition and to recovery sleep condition to characterize the cell-type, specific and shared gene expression responses to sleep loss and recovery in FC tissue of C57 BL/6J mice. 2. Identification of cell-type specific open chromatin induced by sleep deprivation and sleep loss recovery will be made using ATACseq to identify potential enhancer and promoter sites important to the sleep loss response. 3. The role of Ado activation of ADORA1 receptors in the cell-type specific genomic response to sleep loss will be examined using a conditional Adora1 knockout that is unresponsive to Ado and a glial conditional Adk knockout with chronically elevated brain Ado and elevated SWA during waking and sleep. The first aim may provide fundamental insight into the cellular targets of sleep function with respect to cell-type specificity and differential genomic ontology in response to sleep loss. The second aim is designed to identify DNA loci as potential, cell type-specific, enhancer and/or promoter sites. Identification of these sites is important for characterization of genomic pathways activated by SD and further, is a first step towards understanding and prioritizing GWAS identified loci in non-coding DNA regions for future validation. The third aim will test Ado’s role as a mediator of the genomic changes observed in response to sleep loss and whether or not its mediation of homeostatic sleep SWA can be dissociated from the sleep loss transcriptome.