Mechanisms and Functions of MBP mRNA Transport in Oligodendrocytes

少突胶质细胞中 MBP mRNA 转运的机制和功能

• 批准号: 8833702
• 负责人: Meng-Meng Fu
• 金额: $ 2.12万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8833702
• 关键词: 3' Untranslated Regions; Animal Model; Axon; Binding; Biology; Cell physiology; Cells; Cellular biology; Cerebral Palsy; Cultured Cells; Cytoplasmic Granules; Data; Detection; Disease; Distal; Engineering; Imaging Device; Immunoprecipitation; Label; Life; Link; Mass Spectrum Analysis; Messenger RNA; Modification; Molecular; Molecular Motors; Motor; Multiple Sclerosis; Myelin; Myelin Basic Proteins; Neuraxis; Neuroglia; Neurons; Nuclear Export; Oligodendroglia; Organism; Post-Translational Protein Processing; Process; Property; Proteins; Proteomics; RNA; RNA Binding; RNA Splicing; RNA-Binding Proteins; Regulation; Reporter; Scaffolding Protein; Signal Transduction; Site; Small Interfering RNA; System; Time; Translating; Translations; Travel; cell motility; cellular imaging; in vivo; mouse model; mutant; myelination; neuronal cell body; new therapeutic target; novel; protein expression; public health relevance; research study; therapeutic target

DESCRIPTION (provided by applicant): In the central nervous system, oligodendrocytes ensheath and wrap axons in many concentric layers of myelin, a process that is essential for efficient electrical signaling in axons. In diseases such as multiple sclerosis and cerebral palsy, oligodendrocytes fail to form compact myelin and proper distal localization of myelin basic protein (MBP) is absent. MBP is the most highly expressed mRNA in oligodendrocytes by over 10-fold. It is also a special protein who's mRNA must be transported to distal processes of oligodendrocytes before it can be translated. The mechanism and in vivo function of MBP mRNA transport, however, has remained elusive. This project aims to determine whether regulation of MBP mRNA transport at the cargo level is necessary for myelination. In order to visualize MBP mRNA transport in living primary oligodendrocytes, modern imaging tools, such as the RNA-binding reporter protein MS2, will be developed for the detection and analysis of MBP mRNA motility. Furthermore, two independent mass spectrometry approaches will be used to identify proteins in the MBP mRNA granule that are involved in its transport. Regulators of transport are often scaffolding proteins that have the ability to bind to multiple motor proteins. Thus, candidate regulators of MBP mRNA transport will be confirmed via their ability to associate with motor proteins as well as with siRNA knockdown strategies. Finally, two different mouse models will be made to determine whether MBP mRNA transport is necessary for myelination in vivo. The proposed project will elucidate how motor proteins associate with mRNA cargos and identify the molecular mechanism underlying a pivotal process in oligodendrocyte maturation and myelination and will have important implications for the fields of glial cell biology, molecular motors, RNA protein biology, and myelin diseases. Importantly, since distal MBP protein expression and compact myelin formation fail in myelinating diseases, these findings may reveal novel therapeutic targets for stimulating myelination in disease.

描述（由申请人提供）：在中枢神经系统中，少突胶质细胞将轴突包裹并包裹在许多同心髓磷脂层中，这一过程对于轴突中有效的电信号传导至关重要。在多发性硬化症和脑瘫等疾病中， 少突胶质细胞无法形成致密的髓磷脂，并且缺乏髓磷脂碱性蛋白（MBP）的正确远端定位。 MBP 是少突胶质细胞中表达最高的 mRNA，表达量高出 10 倍以上。它也是一种特殊的蛋白质，其 mRNA 必须转运到少突胶质细胞的远端突起才能翻译。然而，MBP mRNA 转运的机制和体内功能仍不清楚。该项目旨在确定在货物水平上调节 MBP mRNA 运输对于髓鞘形成是否是必要的。为了可视化活原代少突胶质细胞中 MBP mRNA 的转运，将开发现代成像工具，例如 RNA 结合报告蛋白 MS2，用于检测和分析 MBP mRNA 运动性。此外，两种独立的质谱方法将用于鉴定 MBP mRNA 颗粒中参与其运输的蛋白质。运输调节剂通常是能够结合多种运动蛋白的支架蛋白。因此，MBP mRNA 运输的候选调节剂将通过其与运动蛋白以及 siRNA 敲低策略相关的能力得到证实。最后，将制作两种不同的小鼠模型来确定 MBP mRNA 转运是否是体内髓鞘形成所必需的。该项目将阐明运动蛋白如何与 mRNA 货物结合，并确定少突胶质细胞成熟和髓鞘形成关键过程的分子机制，并将对神经胶质细胞生物学、分子马达、RNA 蛋白生物学和髓磷脂疾病领域产生重要影响。重要的是，由于髓鞘疾病中远端 MBP 蛋白表达和致密髓磷脂形成失败，这些发现可能揭示刺激疾病中髓鞘形成的新治疗靶点。