New approaches to local translation: spaceSTAMP of proteins synthesized in axons

局部翻译的新方法：轴突中合成的蛋白质的 spaceSTAMP

• 批准号: 7993398
• 负责人: ROSALIND A. SEGAL
• 金额: $ 16.11万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-08 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7993398
• 关键词: Actins; Address; Affect; Antibodies; Autistic Disorder; Axon; Binding Proteins; Biological; Biological Process; Cell physiology; Cells; Cleaved cell; Cognitive; Collaborations; Communication; Communities; Covalent Interaction; Data; Defect; Developmental Delay Disorders; Disease; Distal; Encephalitis; FMRP; Figs - dietary; Fragile X Gene; Fragile X Syndrome; Functional disorder; Goals; Inherited; Lead; Learning; Left; Limbic Encephalitis; Link; Location; Malignant neoplasm of lung; Mental Retardation; Mental disorders; Messenger RNA; Methodology; Methods; Mitochondria; Nervous system structure; Neurons; Paraneoplastic Syndromes; Patients; Peptide Hydrolases; Presynaptic Terminals; Problem Solving; Protein Biosynthesis; Proteins; RNA-Binding Proteins; Role; Shipping; Ships; Stress; Techniques; Testing; Time; Translating; Translations; Work; autistic behaviour; environmental stressor; neuronal cell body; neuropsychiatry; neurotrophic factor; novel strategies; public health relevance; response; transcription factor

DESCRIPTION (provided by applicant): For many years, conventional wisdom declared that no protein translation occurs in the axons of mammalian neurons. Instead all proteins needed for axonal functions were supposed to be synthesized in the cell bodies and shipped out to the axons. More recently, accumulating data has provided evidence that many mRNAs are found in axons in the mammalian nervous system. Evidence indicating that local translation in axons may be regulated by electrical activity, neurotrophic factors, and stress, have led to the hypothesis that axonal translation is particularly important for plasticity during learning, or in response to environmental stressors. In support of this idea, defects in local translation have been linked to Fragile X syndrome, which causes cognitive problems and autistic behaviors, and paraneoplastic disorders, which cause encephalitis in some patients with lung cancers. However, the mechanisms regulating local translation, and why defects in local translation lead to neuro-psychiatric dysfunction, are not understood. In part, this reflects the inadequacy of current approaches for analyzing local protein synthesis or identifying biological functions that rely on regulated local translation. To address this problem we initiated a collaboration with Michael Lin. Michael recently developed a technique called timeSTAMP, to identify proteins that are translated at a particular time period (4). We are working with Michael to modify this approach so it can be used for spaceSTAMP, to tag proteins translated in a particular location, and follow them over time and space. The goal of this proposal is develop the spaceSTAMP approach and use it to ask: Is there regulated local translation in axons? Are such locally translated proteins functionally important? Are locally translated proteins restricted to the axonal compartment, or do they facilitate communication between axonal terminals and remote portions of the neuronal cell body? These studies will develop and test a spaceSTAMP technique that allows one to tag proteins synthesized in axons, and to follow these components in time and space. This approach can then be used by the scientific community to solve problems such as the functional importance of the fragile X gene product, and ways in which activity or neurotrophin regulated translation contribute to neuro-psychiatric disorders. PUBLIC HEALTH RELEVANCE: These studies will develop a new technique that we call spaceSTAMP. This technique allows one to tag proteins made in one part of the cell and to follow these components as they move in time and space. This approach will enable the scientific community to understand diseases that affect local protein synthesis; these diseases include Fragile X syndrome, one of the most common inherited causes of developmental delay and of autism.

描述（由申请人提供）：多年来，传统观点认为哺乳动物神经元的轴突中不发生蛋白质翻译。相反，轴突功能所需的所有蛋白质都应该在细胞体中合成并运送到轴突。最近，越来越多的数据提供了证据表明，许多mRNA被发现在哺乳动物神经系统的轴突。有证据表明，轴突的局部翻译可能受到电活动、神经营养因子和压力的调节，这导致了轴突翻译对学习过程中的可塑性或对环境压力的反应特别重要的假设。为了支持这一观点，局部翻译的缺陷与脆性X综合征有关，脆性X综合征会导致认知问题和自闭症行为，而副肿瘤性疾病会导致一些肺癌患者的脑炎。然而，调节局部翻译的机制，以及为什么局部翻译缺陷导致神经精神功能障碍，还不清楚。在某种程度上，这反映了目前分析局部蛋白质合成或鉴定依赖于受调控的局部翻译的生物功能的方法的不足。为了解决这个问题，我们开始与Michael Lin合作。Michael最近开发了一种名为timeSTAMP的技术，用于识别在特定时间段翻译的蛋白质。我们正在与Michael合作修改这种方法，以便它可以用于spaceSTAMP，标记在特定位置翻译的蛋白质，并随时间和空间跟踪它们。这项提议的目标是发展spaceSTAMP方法，并使用它来问：轴突中是否存在受调节的局部翻译？这些局部翻译的蛋白质在功能上重要吗？局部翻译的蛋白质是局限于轴突区室，还是促进轴突终末和神经元细胞体远端部分之间的通讯？这些研究将开发和测试一种spaceSTAMP技术，该技术允许标记轴突中合成的蛋白质，并在时间和空间上跟踪这些成分。然后，科学界可以使用这种方法来解决诸如脆性X基因产物的功能重要性，以及活动或神经营养因子调节翻译导致神经精神疾病的方式等问题。 公共卫生相关性：这些研究将开发一种新技术，我们称之为spaceSTAMP。这项技术允许人们标记细胞一部分中产生的蛋白质，并跟踪这些成分在时间和空间中的移动。这种方法将使科学界能够了解影响局部蛋白质合成的疾病;这些疾病包括脆性X综合征，这是发育迟缓和自闭症最常见的遗传原因之一。