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: 8240550
• 负责人: ROSALIND A. SEGAL
• 金额: $ 41.91万
• 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/8240550
• 关键词: Actins; Address; Affect; Antibodies; Autistic Disorder; Axon; Binding Proteins; Biological; Biological Process; Cell physiology; Cells; Cleaved cell; Cognitive; Collaborations; Communication; Communities; Data; Defect; Developmental Delay Disorders; Disease; Distal; Encephalitis; FMRP; 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，传统的智慧宣布哺乳动物神经元的轴突中没有蛋白质翻译。取而代之的是，轴突功能所需的所有蛋白质都应在细胞体中合成并运送到轴突中。最近，累积数据提供了证据，表明在哺乳动物神经系统的轴突中发现了许多mRNA。表明轴突中局部翻译的证据可能受到电活动，神经营养因素和压力的调节，这导致假设是，轴突翻译对于学习过程中的可塑性或对环境压力源的响应尤为重要。为了支持这一想法，局部翻译中的缺陷与脆弱的X综合征有关，X综合征会导致认知问题和自闭症行为，以及副肿瘤疾病，这些疾病会导致一些肺癌患者的脑炎。然而，尚不清楚调节局部翻译的机制，以及为什么局部翻译中的缺陷导致神经精神上的功能障碍。在某种程度上，这反映了目前用于分析局部蛋白质合成或识别依赖于调节局部翻译的生物学功能的方法不足。为了解决这个问题，我们与迈克尔·林（Michael Lin）进行了合作。迈克尔最近开发了一种称为时间戳的技术，以识别在特定时间段内翻译的蛋白质（4）。我们正在与Michael合作修改这种方法，以便它可用于在特定位置中翻译的蛋白质，并在时间和空间上跟随它们。该提案的目的是开发Spacestamp方法并使用它来提出：轴突中是否有监管的本地翻译？如此局部翻译的蛋白质在功能上是否重要？是局部翻译的蛋白质是否仅限于轴突室，还是促进神经元细胞体的轴突末端和偏远部分之间的通信？这些研究将开发和测试一种允许轴突中合成的蛋白质并在时间和空间中遵循这些组件的蛋白质的水上管技术。然后，科学界可以使用这种方法来解决诸如脆弱X基因产物的功能重要性，以及活动或神经营养蛋白调节翻译有助于神经精神疾病的方法。 公共卫生相关性：这些研究将开发一种我们称为Spacestamp的新技术。该技术使人们可以在细胞的一部分中标记蛋白质，并在这些组件上移动时，并跟随它们在时空移动。这种方法将使科学界能够理解影响局部蛋白质合成的疾病。这些疾病包括脆弱的X综合征，X综合征是发育延迟和自闭症的最常见原因之一。