Translational control of a complete developing sensory circuit

完整发育中的感觉回路的平移控制

• 批准号: 10678098
• 负责人: Ryan E Loker
• 金额: $ 6.91万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10678098
• 关键词: Address; Adult; Atlases; Bioinformatics; Biological Assay; Biological Models; Birth; Caring; Cell physiology; Cells; Chromatin; Development; Drosophila genus; Engineering; Epigenetic Process; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Transcription; Genome; Genomics; Glutamates; Growth; Health; Homeostasis; Human Development; Impairment; Individual; Invertebrates; Ion Channel; Link; Machine Learning; Messenger RNA; Methods; MicroRNAs; Modeling; Molecular; Morphology; Nervous System; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Neurosciences; Neurotransmitters; Optic Lobe; Organ; Pathway interactions; Pattern; Physiology; Post-Transcriptional Regulation; Process; Protein Biosynthesis; Proteins; RNA; RNA-Binding Proteins; RNA-Protein Interaction; Regulation; Repression; Reproducibility; Research; Role; Sensory; Shapes; Signal Pathway; Specific qualifier value; Subcellular Spaces; Synapses; System; Technical Expertise; Techniques; Testing; Therapeutic; Tissues; Training; Transcript; Translating; Translational Regulation; Translational Repression; Translations; Vertebrates; Visual System; Work; Writing; axon guidance; cell type; cholinergic; data dissemination; derepression; design; experimental study; fly; genome-wide; genome-wide analysis; improved; insight; mRNA Translation; nervous system development; neural; neurodevelopment; neurotransmission; ribosome profiling; sensory system; single-cell RNA sequencing; technological innovation; tool; transcriptome; transcriptomics

Project Summary Development of the nervous system requires the generation of diverse neuronal types that subsequently drastically alter their shape and physiology to form vast interconnected networks. Each process requires precise gene regulation. Technological innovations have drastically increased the scale at which we can distinguish neuronal identities or stages based on transcriptomics or epigenetics. However, principles guiding post-transcriptional control of gene expression, which is essential for neural development and homeostasis, by regulatory factors including RNA-binding proteins (RBP) and miRNAs has not been defined at a similar systematic level. As numerous neurodegenerative disorders have been linked to impaired RBP-RNA interactions, and there is a huge need for improved cell type engineering strategies for therapeutics, insight into this process is critically important. I propose to use the Drosophila visual system as a model to study post- transcriptional regulation during specification and wiring of an entire neural sensory system. Specifically, this proposal focuses on the regulation of mRNA translational repression, which is a conserved feature of both neuronal fate diversification and differentiation in vertebrate and invertebrates. A single-cell transcriptomic atlas of the developing visual system, or optic lobe, was recently described which defines the transcriptome of each neuronal type in the optic lobe throughout development. Consistent with results from other model systems, many genes associated with terminal neuronal function were detected at transcripts in the immature neurons well before they are functionally required, and the corresponding protein was absent for two selected genes studied. I aim to define the scope of translational regulation in this system by first performing whole-tissue and select cell-specific ribosome profiling of the optic lobe over development, and determine the upstream control of this process using a combination of bioinformatics and genetics (Aim 1). Next I will tease apart the molecular and cellular mechanism by which two genes associated with different neuron signaling pathways are repressed at the translational level to assess how their expression is coordinated (Aim 2). Finally, I will adapt a single-cell translation profiling technique, scRibo-STAMP, in the Drosophila visual system to profile translation of all optic lobe neurons during specification and wiring. Combined with machine-learning based analysis, I will predict RBP/miRNA-RNA target interactions to gain fundamental insight into how RNA regulatory networks are shaped (Aim 3). Together this study will provide significant insights into the role of translational regulation during formation of the nervous system relevant to both human development and health.

项目概要 神经系统的发育需要产生不同的神经元类型，随后 彻底改变它们的形状和生理机能，形成巨大的互连网络。每个过程都需要 精准的基因调控。技术创新极大地扩大了我们的规模 根据转录组学或表观遗传学区分神经元身份或阶段。然而，指导原则 基因表达的转录后控制，这对于神经发育和稳态至关重要 包括 RNA 结合蛋白 (RBP) 和 miRNA 在内的调控因子尚未在类似的研究中得到定义。 系统化水平。由于许多神经退行性疾病与 RBP-RNA 受损有关 相互作用，并且非常需要改进用于治疗、洞察力的细胞类型工程策略 进入这个过程至关重要。我建议使用果蝇视觉系统作为模型来研究后处理 整个神经感觉系统的规范和连接过程中的转录调节。具体来说，这 提案重点关注 mRNA 翻译抑制的调节，这是两者的保守特征 脊椎动物和无脊椎动物的神经元命运多样化和分化。单细胞转录组图谱 最近描述了发育中的视觉系统或视叶的转录组，它定义了每个视觉系统的转录组 整个发育过程中视叶的神经元类型。与其他模型系统的结果一致， 在未成熟神经元的转录本中检测到许多与终末神经元功能相关的基因 早在它们被功能需要之前，两个选定的基因就缺乏相应的蛋白质 研究过。我的目标是通过首先进行全组织和 选择发育过程中视叶的细胞特异性核糖体分析，并确定上游控制 结合生物信息学和遗传学来研究这一过程（目标 1）。接下来我将梳理一下分子 以及与不同神经元信号通路相关的两个基因被抑制的细胞机制 在翻译水平上评估它们的表达是如何协调的（目标 2）。最后，我将改编一个单细胞 果蝇视觉系统中的平移分析技术 scRibo-STAMP 可分析所有光学系统的平移 叶神经元在规范和接线过程中。结合基于机器学习的分析，我将预测 RBP/miRNA-RNA 靶标相互作用，以获得有关 RNA 调控网络如何形成的基本见解 （目标 3）。总之，这项研究将为翻译调控在过程中的作用提供重要的见解。 神经系统的形成与人类发育和健康相关。