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.

项目摘要 神经系统的发展需要产生不同类型的神经元，随后 彻底改变了它们的形状和生理，形成了巨大的相互连接的网络。每个流程都需要 精确的基因调控。技术创新极大地增加了我们可以达到的规模 根据转录学或表观遗传学区分神经元的身份或阶段。然而，指导原则 转录后对基因表达的控制，这是神经发育和动态平衡所必需的，通过 包括RNA结合蛋白(RBP)和miRNAs在内的调控因子还没有被定义为类似的 系统层面。由于许多神经退行性疾病与RBP-RNA受损有关 相互作用，以及对于治疗学、洞察力的改进的细胞类型工程策略的巨大需求 进入这一进程是至关重要的。我建议用果蝇的视觉系统作为研究后视觉系统的模型。 在整个神经感觉系统的指定和连接过程中的转录调节。具体地说，这 提案的重点是调节mrna的翻译抑制，这是两者的保守特征。 脊椎动物和无脊椎动物神经元命运的多样化和分化。一种单细胞转录图谱 最近描述了发育中的视觉系统，或称视叶，它定义了每一个 视叶发育过程中的神经元型。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。 早在功能需要它们之前，相应的蛋白质在两个选定的基因中就缺失了 学习。我的目标是在这个系统中通过首先执行整体组织和 选择视叶发育过程中的细胞特异性核糖体图谱，并确定上游调控 利用生物信息学和遗传学的结合来研究这一过程(目标1)。接下来，我将梳理分子 和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 在翻译层面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 果蝇视觉系统中的翻译简档技术，scRibo-Stamp，以简档所有视神经的翻译 脑叶神经元在规范和连接过程中。结合基于机器学习的分析，我将预测 RBP/miRNA-RNA靶向相互作用，以获得对RNA调控网络如何形成的基本见解 (目标3)。综上所述，这项研究将为翻译调控在 与人类发育和健康相关的神经系统的形成。