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，或者说是允许的，睡眠需求的解决。睡眠需要的解决反映了睡眠功能 睡眠稳态（sleep homeostasis）现在对睡眠/觉醒状态的控制了解得很多 表达和持续时间，但睡眠功能不太清楚。 基因的显著差异表达（DEG's;额叶皮质大块组织中表达6000/13000; FC） 已经观察到对6小时睡眠剥夺（SD）的响应。对于CNS组织特别重要， 睡眠调节基因本体支持神经元兴奋性和传递的睡眠控制。 细胞功能的睡眠依赖性调节可能会根据不同细胞类型的特定作用而变化 然而，人们对睡眠DEG的细胞类型特异性及其对生物学内和 负责SD反应的细胞间通路。 Ado可能涉及作为睡眠需要的细胞间（神经胶质到神经元）介导剂，因为其受体被激活， ADORA 1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 还没有被检查。 提出了以下具体目标来表征睡眠丧失的基因组反应， DEG的细胞类型特异性和在细胞间和细胞外决定其SD反应的途径， 细胞内水平，在小鼠FC。 1.将比较睡眠丧失后细胞类型特异性mRNA表达的差异表达 自由睡眠状态和恢复睡眠状态，以表征细胞类型、特异性和共享性 C57 BL/6 J小鼠FC组织中对睡眠丧失和恢复的基因表达反应。 2.睡眠剥夺和睡眠恢复诱导的细胞类型特异性开放染色质的鉴定将是 使用ATACseq来鉴定对睡眠丧失反应重要的潜在增强子和启动子位点。 3. Ado激活ADORA 1受体在睡眠丧失的细胞类型特异性基因组应答中的作用将 使用对Ado无反应的条件Adora 1敲除和神经胶质条件Adk 在清醒和睡眠期间具有慢性升高的脑Ado和升高的SWA。 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。 特异性和差异基因组本体论的研究。第二个目标是确定 DNA基因座作为潜在的细胞类型特异性增强子和/或启动子位点。这些网站的识别是 重要的是表征的基因组途径激活SD和进一步，是第一步， 理解并优先考虑GWAS在非编码DNA区域中鉴定的基因座，以供将来验证。第三 aim将测试Ado作为睡眠不足引起的基因组变化的调节剂的作用，以及Ado是否 是否其调节稳态睡眠的SWA可以与睡眠丧失转录组分离。