Neuronal Activity-dependent Pomoter Usage

神经元活动依赖性启动器的使用

• 批准号: 10451349
• 负责人: Shigeki Iwase
• 金额: $ 22.88万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10451349
• 关键词: Alternative Splicing; Animals; Behavior; Biological; Brain; Bromouridine sequencing; Cells; Cognition Disorders; Communication; Complementary DNA; Coupled; Cyclic AMP; Cyclic GMP; Data; Destinations; Detection; Disease; Electrophysiology (science); Environment; Epitopes; Future; Gene Expression Profile; Gene Expression Profiling; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genomics; Goals; Housing; Human; Impairment; Length; Libraries; Ligation; Light; Mental Depression; Mental disorders; Methods; Mitochondria; Modeling; Molecular; Mus; Nature; Neuroglia; Neurons; Nuclear RNA; Outcome; Peptide Signal Sequences; Play; Population; Post-Translational Protein Processing; Process; Prosencephalon; Protein Isoforms; Proteins; Proteome; RNA; RNA Binding; Research; Ribosomal Proteins; Ribosomes; Role; Schizophrenia; Second Messenger Systems; Sensory; Signal Transduction; Specificity; Stimulus; Structure; Synapses; Synaptic plasticity; Testing; Transcript; Transcription Initiation Site; autism spectrum disorder; base; cell type; cognitive function; deep sequencing; excitatory neuron; experience; experimental study; extracellular; genome-wide; improved; in vivo; inhibitory neuron; insight; knock-down; mRNA Expression; mouse model; neural circuit; neural network; neurogenetics; novel; phenomenological models; phosphoric diester hydrolase; polypeptide; promoter; relating to nervous system; response; sensory input; transcriptome; transcriptome sequencing; translatome

ABSTRACT Experience-dependent remodeling of neural circuits enables animals to adapt their behavior to ever- changing environments. Neuronal activity-dependent molecular alterations, elicited by sensory inputs, at all levels from transcription to post-translational modifications are the basis of neural rewiring. Not surprisingly, impaired molecular responses to neuronal activity are a common hallmark of a broad range of cognitive disorders, such as depression, schizophrenia, and autism. Thus, comprehending activity-dependent molecular changes is crucial to understanding the mechanisms of these mental disorders. Over the past several years, extensive studies have made excellent strides in profiling activity-dependent changes in the transcriptome, alternative splicing, and proteome. These studies have provided significant new insights into how the brain integrates sensory inputs and reorganizes specific neural networks. However, a portion of the RNA molecule has escaped genome-wide scrutiny―the 5'-ends. Widely-used RNA-seq methods underrepresent the 5'-ends of RNAs. Existing methods to profile transcription start sites (TSS), however, predominantly represent the 5' ends; therefore, most of the transcriptome is absent in such libraries, making it challenging to connect TSS usage to dynamic mRNA expression. We recently developed a full-length nascent-transcriptome profiling method, Bru-seq-DLAF, which combines sensitive detection of TSS and nascent RNA sequencing. With Bru-seq-DLAF, we have unveiled tens of genes that dynamically change TSS upon reducing or increasing network activity. Most of these TSS led to alterations in polypeptides at their amino termini. Furthermore, the protein isoforms were conserved between mice and humans, indicating that activity-dependent TSS usage may represent a conserved new layer of gene regulation underlying synaptic plasticity. The proposed research goals are to 1) thoroughly characterize this novel phenomenon in mouse models and 2) determine the roles of activity-dependent TSS in synaptic plasticity. We hypothesize that neuronal activity-dependent TSS controls synaptic plasticity by changing the subcellular localization of proteins. Our team is uniquely poised to test this hypothesis with its unified expertise in genomics and synaptic plasticity. A positive outcome of the proposed study is to illuminate activity-dependent TSS selection as an intricate molecular mechanism for synaptic plasticity. To our knowledge, our data represent the first genome- wide characterization of TSS alterations upon extracellular stimuli in animal cells. Furthermore, many genes that undergo activity-dependent TSS selection have been implicated in mental disorders such as schizophrenia and autism spectrum disorders. Thus, the proposed study will improve the genetics of mental disorders and guide future functional studies on disease-associated genes.

抽象的 依赖经验的神经回路的重塑使动物能够使其行为适应不断 改变环境。神经元活性依赖性分子改变，从感觉输入引起 从转录到翻译后修饰的水平是神经重新启动的基础。毫不奇怪， 分子对神经元活性的响应受损是广泛认知的常见标志 抑郁症，精神分裂症和自闭症等疾病。理解活动依赖性分子 变化对于理解这些精神障碍的机制至关重要。在过去的几年中， 广泛的研究在转录组的分析依赖性变化方面取得了长足的进步， 替代剪接和蛋白质组。这些研究为大脑如何如何 集成感官输入并重新组织特定的神经网络。 但是，RNA分子的一部分已经逃脱了全基因组的审查5'末端。广泛使用 RNA-seq方法不足以占RNA的5'端。现有的介绍转录开始站点的方法 （TSS）但是，主要代表5'末端；因此，大多数转录组都没有 图书馆，使TSS使用率连接到动态mRNA表达的挑战。 我们最近开发了一种全长的新生转录组分析方法Bru-Seq-DLAF，该方法 结合了TSS和新生RNA测序的敏感检测。使用Bru-Seq-Dlaf，我们已经揭幕了 减少或增加网络活性时动态改变TSS的数十个基因。这些TSS大多数 导致其氨基末端的多肽改变。此外，蛋白质同工型是保守的 在小鼠和人类之间，表明活动依赖性TSS的使用可能代表一种新的 基因调节层的基础突触可塑性。拟议的研究目标是1）彻底 在小鼠模型中表征这种新现象，2）确定活性依赖性TSS的作用 突触可塑性。我们假设神经元活动依赖性TSS控制着通过 改变蛋白质的亚细胞定位。我们的团队被毒死了，以检验这一假设 基因组学和突触可塑性方面的统一专业知识。 拟议的研究的积极结果是将活动依赖性TSS选择作为一种 复杂的分子机制，用于突触可塑性。据我们所知，我们的数据代表了第一个基因组 - 在动物细胞中细胞外刺激时TSS改变的广泛表征。此外，许多基因 在精神分裂症等精神障碍中，已实施了与活动有关的TSS选择 和自闭症谱系障碍。这是拟议的研究将改善精神障碍的遗传学和 指导与疾病相关基因的未来功能研究。