A Fast Proteometric Synapse Census Platform

快速蛋白质突触普查平台

• 批准号: 8460319
• 负责人: Stephen J Smith
• 金额: $ 21.78万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-12 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8460319
• 关键词: Accounting; Address; Adult; Animal Experimentation; Antibodies; Architecture; Biological Assay; Brain; Brain Diseases; Censuses; Central Nervous System Diseases; Classification; Clinical; Computer software; Data; Development; Disease; Distress; Face; Hour; Human; Image; Immunofluorescence Immunologic; In Situ; Individual; Journals; Knowledge; Label; Left; Link; Measures; Memory; Mental disorders; Methods; Microscope; Molecular; Mus; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurons; Neurosciences; Neurotransmitters; Population; Property; Proteins; Protocols documentation; Publications; Publishing; Reproducibility; Research; Resolution; Retrieval; Specificity; Specimen; Speed; Standardization; Structure; Surveys; Synapses; System; Technology; Time; Work; barrel cortex; base; design; improved; information framework; innovation; memory encoding; molecular imaging; neural circuit; novel; open source; postsynaptic; presynaptic; prevent; sensor; software development; tomography; tool; wiki

DESCRIPTION (provided by applicant): Synapses of the mammalian central nervous system (CNS) are very deeply diverse in both molecular and functional properties. At present, unfortunately, our understanding of this diversity is rudimentary, and quantitative data on the subject are very few. Left unfathomed, CNS synapse diversity poses formidable obstacles to better understanding of the development, function and disorders of the brain's synaptic circuitry. The major reason for the persistence of this distressing state of ignorance lies in the fact that tools for exploring synapse populations at the level of individual synapses are few and limited in their capabilities. To address the challenges synapse diversity poses to both basic and clinical neuroscience, this project aims to develop a superlative new proteometric imaging platform capable of analyzing very large synapse populations in situ with single- synapse resolution. Deployment and dissemination of this platform will facilitate study and treatment of the many neurodevelopmental, mental and neurodegenerative disorders linked to specific synapse subpopulations, as well as opening new perspectives on molecular mechanisms, circuit architectures and disorders of CNS memory encoding, storage and retrieval. The platform will be based on immunofluorescence array tomography (IAT) and involve development of novel antibody standardization protocols and novel image acquisition hardware and software. These innovations will improve the reproducibility, quantitative reliability, and speed of IAT by large margins and overcome limitations that have so far prevented proteometric analysis of large synapse populations at the single-synapse level. The platform would be capable of proteometric census of a million of more synapses per hour, at 50 or more markers per synapse, while maintaining precise neuroanatomical and molecular coordinates for each synapse. The new platform will be demonstrated by a 7-marker proteometric survey of an adult mouse cortex barrel column that would enumerate each of the tens of millions of synapses in the column and allow classification of each synapse based on neurotransmitter type and a set of neurons type markers. The results will be disseminated via methods publications, open-source PUBLIC HEALTH RELEVANCE: Abnormalities of individual synapses and of the brain's large and diverse synapse populations are widely believed to account for many or most neurodevelopmental, neurodegenerative and substance-abuse-related brain disorders. This work will address major gaps in present knowledge of such abnormalities by developing superlative new tools for the very rapid and highly detailed quantitative survey of large synapse populations in both research animal and human brain specimens.

描述（由申请人提供）：哺乳动物中枢神经系统（CNS）的突触在分子和功能特性上都非常多样化。不幸的是，目前我们对这种多样性的理解还很初级，关于这个主题的定量数据也很少。如果不了解中枢神经系统突触多样性，就会给更好地理解大脑突触回路的发育、功能和紊乱带来巨大的障碍。这种令人痛苦的无知状态持续存在的主要原因在于，用于在单个突触水平上探索突触群体的工具很少且能力有限。为了解决突触多样性给基础和临床神经科学带来的挑战，该项目旨在开发一种最高级的新型蛋白质成像平台，能够以单突触分辨率原位分析非常大的突触群体。该平台的部署和传播将促进与特定突触亚群相关的许多神经发育、精神和神经退行性疾病的研究和治疗，并为中枢神经系统记忆编码、存储和检索的分子机制、电路结构和疾病开辟新的视角。该平台将基于免疫荧光阵列断层扫描（IAT），涉及新型抗体标准化协议和新型图像采集硬件和软件的开发。这些创新将大幅提高 IAT 的可重复性、定量可靠性和速度，并克服迄今为止阻碍在单突触水平上对大型突触群体进行蛋白质计量分析的限制。该平台能够每小时对一百万个以上突触进行蛋白质普查，每个突触有 50 个或更多标记，同时保持每个突触精确的神经解剖学和分子坐标。新平台将通过对成年小鼠皮层桶状柱进行 7 标记蛋白质测量研究来展示，该调查将枚举柱中数千万个突触中的每一个，并允许根据神经递质类型和一组神经元类型标记对每个突触进行分类。结果将通过方法出版物、开源 公共健康相关性：人们普遍认为，个体突触和大脑大量多样的突触群的异常是许多或大多数神经发育、神经退行性和药物滥用相关脑部疾病的原因。这项工作将通过开发最先进的新工具来对研究动物和人脑标本中的大量突触群体进行非常快速和高度详细的定量调查，从而解决目前对此类异常的认识的主要差距。