Neuronal Activity-dependent Pomoter Usage

神经元活动依赖性启动器的使用

• 批准号: 10591519
• 负责人: Shigeki Iwase
• 金额: $ 18.95万
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10591519
• 关键词: Alternative Splicing; Animals; Behavior; Biological; Brain; Bromouridine sequencing; Cells; Cognition Disorders; Communication; Complementary DNA; Coupled; Cyclic AMP; Cyclic GMP; Darkness; 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; 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; neural circuit; neural network; neurogenetics; novel; phenomenological models; phosphoric diester hydrolase; polypeptide; promoter; response; sensory input; sensory integration; transcriptome; transcriptome sequencing; transcriptomic profiling; 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 依赖于经验的神经回路重塑使动物能够适应自己的行为 不断变化的环境。由感觉输入引起的神经元活动依赖性分子改变 从转录到翻译后修饰的水平是神经重新布线的基础。毫不奇怪， 对神经元活动的分子反应受损是广泛认知的共同标志 疾病，例如抑郁症、精神分裂症和自闭症。因此，理解活性依赖性分子 变化对于理解这些精神障碍的机制至关重要。在过去的几年里， 广泛的研究在转录组中活性依赖性变化的分析方面取得了巨大的进步， 选择性剪接和蛋白质组。这些研究为大脑如何运作提供了重要的新见解。 整合感觉输入并重组特定的神经网络。 然而，RNA 分子的一部分（即 5' 端）却逃脱了全基因组范围的审查。广泛使用 RNA-seq 方法未能充分代表 RNA 的 5' 端。分析转录起始位点的现有方法 然而，(TSS) 主要代表 5' 端；因此，大部分转录组在此类中不存在 库，使得将 TSS 使用与动态 mRNA 表达联系起来具有挑战性。 我们最近开发了一种全长新生转录组分析方法 Bru-seq-DLAF，该方法 结合了 TSS 的灵敏检测和新生 RNA 测序。通过 Bru-seq-DLAF，我们揭开了 数十个基因在减少或增加网络活动时动态改变 TSS。其中大部分 TSS 导致多肽氨基末端的改变。此外，蛋白质亚型是保守的 小鼠和人类之间的差异，表明依赖于活动的 TSS 使用可能代表了一种保守的新机制 突触可塑性的基因调控层。拟议的研究目标是 1) 彻底 在小鼠模型中描述这一新现象，并2）确定活动依赖性 TSS 在 突触可塑性。我们假设神经元活动依赖性 TSS 通过以下方式控制突触可塑性： 改变蛋白质的亚细胞定位。我们的团队准备用其独特的方法来检验这一假设 基因组学和突触可塑性方面的统一专业知识。 拟议研究的一个积极成果是阐明活动依赖性 TSS 选择作为一种 突触可塑性的复杂分子机制。据我们所知，我们的数据代表了第一个基因组- 动物细胞中细胞外刺激引起的 TSS 变化的广泛表征。此外，许多基因 进行活动依赖性 TSS 选择与精神分裂症等精神障碍有关 和自闭症谱系障碍。因此，拟议的研究将改善精神障碍的遗传学和 指导未来疾病相关基因的功能研究。