The Regulation and Function of Neuron-Specific Alternative Splicing

神经元特异性选择性剪接的调控和功能

• 批准号: 10318594
• 负责人: Sika Zheng
• 金额: $ 34.49万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-15 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318594
• 关键词: Address; Alleles; Alternative Splicing; Axon; Biochemical; Biological Assay; Birth; Brain; Brain Diseases; CRISPR/Cas technology; Cell Death; Cell physiology; Cells; Cellular biology; Code; Collaborations; Computational Biology; Cues; Data; Defect; Development; Down-Regulation; Embryo; Event; Evolution; Exhibits; Gene Expression; Genes; Genetic; Genetic Determinism; Genetic Programming; Goals; Hippocampus (Brain); In Vitro; Knock-out; Mediating; Messenger RNA; Methods; Microtubules; Modeling; Molecular; Morphogenesis; Morphology; Mus; Mutant Strains Mice; Mutation; Neurobiology; Neurodevelopmental Disorder; Neuronal Differentiation; Neurons; Nuclear RNA; Phenotype; Physiological; Play; Polypyrimidine Tract-Binding Protein; Positioning Attribute; Protein Isoforms; Proteins; Proteome; Public Health; RNA Splicing; RNA-Binding Proteins; Recording of previous events; Regenerative Medicine; Regulation; Research; Resources; Risk Factors; Role; Signal Transduction; Synapses; System; Testing; Time; Tissues; Variant; Vertebral column; base; extracellular; genetic manipulation; human disease; in vivo; insight; loss of function; mRNA Precursor; nervous system disorder; neural circuit; prevent; repaired; synaptogenesis; tool; transcription factor; transcriptome

Given the increasing evidence that RNA binding proteins (RBPs) serve as genetic causes or predisposing risk factors underlying a wide spectrum of neurological diseases, a pressing need exists to understand their specific cellular functions in developing and mature brains. Similar to transcription factors, RBPs could be master regulators of certain cellular processes owing to their coordinated target sets. Unlike widely-studied transcription factors, the physiological functions of RBPs are historically underexplored. Most of the dozen RBPs found to exhibit tissue-specific expression are involved in regulating neuronal alternative splicing. We and others have revealed the large-scale genetic programming of alternative splicing during embryonic brain development, resulting in neuron-specific alternative isoforms in mature brains. We show many of these splicing changes are mediated by a nuclear RBP, polypyrimidine tract-binding protein 2 (PTBP2). PTBP2 exhibits dynamic temporal expression during neuronal differentiation and may orchestrate the developmental programming of alternative splicing in order to accomplish the prolonged and continuous neuronal morphological transformations. Our previous study shows that downregulation of PTBP2 prior to synaptogenesis is necessary for spine formation. Our newest data show that PTBP2 has additional functions in early differentiating neurons prior to synapse formation. Guided by strong preliminary data, we hypothesize that PTBP2 governs neuron-specific splicing to control the initiation of neuronal polarity. This proposal will address multiple critical barriers to the study of neuronal polarity and provide a new framework for functional analysis of alternative splicing. Our long history of researching alternative splicing and PTBP2 in the brain places us in a unique position to advance these fields. Our team, with complementary expertise in genetics, neurobiology, and molecular cellular biochemical and computational biology, has demonstrated successful collaborations. We have generated new tools and resource to thoroughly determine the cellular and molecular defects caused by Ptbp2 knockout in cortical neurons. Completion of this project will allow us to further our long-term goal of revealing new genetic, molecular and cellular controls of neuronal polarity and morphogenesis that enables neural circuit formation.

鉴于越来越多的证据表明 RNA 结合蛋白 (RBP) 是遗传原因或诱发风险的因素 多种神经系统疾病的潜在因素，迫切需要了解它们 发育中和成熟大脑中的特定细胞功能。与转录因子类似，RBP 可以 由于其协调的目标集，某些细胞过程的主要调节者。与广泛研究的不同 作为转录因子，RBP 的生理功能历来未被充分研究。十几个中的大多数 发现表现出组织特异性表达的 RBP 参与调节神经元选择性剪接。我们 等人揭示了胚胎大脑中选择性剪接的大规模遗传编程 发育，导致成熟大脑中神经元特异性的替代亚型。我们展示了其中许多 剪接变化是由核 RBP、聚嘧啶束结合蛋白 2 (PTBP2) 介导的。 PTBP2 在神经元分化过程中表现出动态的时间表达，并可能协调发育 选择性剪接的编程以实现长期且连续的神经元 形态转变。我们之前的研究表明，PTBP2 的下调之前 突触发生对于脊柱的形成是必要的。我们的最新数据表明 PTBP2 在以下方面具有附加功能： 突触形成之前的早期分化神经元。在强有力的初步数据的指导下，我们假设 PTBP2 控制神经元特异性剪接以控制神经元极性的启动。该提案将 解决神经元极性研究的多个关键障碍，并为功能性研究提供新的框架 选择性剪接分析。我们研究大脑选择性剪接和 PTBP2 的悠久历史 使我们处于推动这些领域发展的独特地位。我们的团队在遗传学方面具有互补的专业知识， 神经生物学、分子细胞生化和计算生物学已被证明是成功的 合作。我们已经开发出新的工具和资源来彻底确定细胞和分子 皮质神经元中 Ptbp2 敲除引起的缺陷。该项目的完成将使我们能够进一步 揭示神经元极性和细胞极性的新遗传、分子和细胞控制的长期目标 能够形成神经回路的形态发生。