A genomic characterization of the response to sleep loss

睡眠不足反应的基因组特征

• 批准号: 10928421
• 负责人: Robert W Greene
• 金额: $ 41万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-21 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10928421
• 关键词: ADORA1 gene; ATAC-seq; Adenosine; Adenosine A1 Receptor; Adenosine Kinase; Affinity; Attenuated; Binding; Binding Sites; Biological; Brain; Cell physiology; Cells; Chromatin; Chronic; DNA; DNA Binding; Dissociation; Enhancers; Foundations; Future; Gene Expression; Genes; Genomics; Growth; Homeostasis; Hour; Knock-out; Mediation; Mediator; Mental disorders; Metabolic syndrome; Mitochondria; Molecular; Mus; Nerve Degeneration; Neurobiology; Neuroglia; Neurons; Non-Essential Amino Acid; Nucleic Acids; Ontology; Pathologic; Pathology; Pathway interactions; Process; Proteins; Purinergic P1 Receptors; Reaction; Receptor Activation; Recovery; Research; Resolution; Ribosomes; Role; Signal Pathway; Signal Transduction; Site; Sleep; Sleep Deprivation; Sleep disturbances; Slow-Wave Sleep; Specificity; Synapses; Testing; Therapeutically Targetable; Time; Tissue-Specific Gene Expression; Tissues; Untranslated RNA; Validation; Work; awake; cell type; cellular targeting; design; differential expression; extracellular; frontal lobe; genome wide association study; insight; mRNA Expression; mutant; myocyte-specific enhancer-binding-factor 2C; neuronal excitability; neurotransmission; permissiveness; promoter; receptor; response; sleep regulation; transcription factor; transcriptome

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.

睡眠门或允许的睡眠需求。睡眠需求的分辨率反映了睡眠功能 响应醒来（睡眠体内平衡）引起的诱导。现在对睡眠/唤醒状态的控制知之甚少 表达和持续时间，但睡眠功能不太了解。 基因的显着差异表达（DEG； 6000/13000在额叶皮层中表达的6000/13000； FC） 响应于6小时的睡眠剥夺（SD）。对于中枢神经系统组织特别重要， 睡眠调制基因本体学支持神经元令人兴奋和传播的睡眠控制。 细胞功能的睡眠依赖性调制可能会根据不同的细胞类型而有所不同 然而，关于睡眠DEG的细胞类型特异性及其对生物学内和生物内的影响很少。 负责SD响应的细胞间途径。 自从其接收器激活以来 Adora1是睡眠式慢波活性（SWA）所必需的，但其在与睡眠相关的基因表达中的作用 尚未检查。 提出了以下具体目的，以表征相对于睡眠损失的基因组反应 DEG的细胞类型特异性和确定其SD响应在细胞间和 小鼠FC中的细胞内水平。 1.将比较细胞类型特异性mRNA表达对睡眠衰退的差异表达 达到脂肪睡眠状况和恢复睡眠状况，以表征细胞类型，特定和共享 基因表达对C57 BL/6J小鼠FC组织的睡眠丧失和恢复的反应。 2。鉴定由睡眠剥夺和睡眠丧失恢复引起的细胞类型特异性开放染色质将是 使用AtacSeq确定对睡眠损失反应很重要的潜在增强子和启动子站点。 3。adora1受体在细胞类型对睡眠丧失的特异性基因组反应中的作用将 使用条件adora1敲除对ADO和有条件ADK的有条件敲除检查 在醒来和睡眠期间，长期升高的大脑ADO和SWA升高的淘汰赛。 第一个目的可以提供对细胞类型的睡眠功能细胞靶标的基本见解 响应睡眠损失的特异性和差异基因组本体论。第二个目标旨在识别 DNA基因座作为电势，细胞类型特异性，增强子和/或启动子位点。这些站点的识别是 对于通过SD激活的基因组途径表征和进一步的基因组途径很重要，这是迈向的第一步 理解和优先考虑GWAS在非编码DNA区域中确定了未来验证的基因座。第三 AIM将测试ADO作为响应睡眠损失的基因组变化的中介的作用 它的稳态睡眠SWA的介导与睡眠损失转录组可以分离。