Defining the translational landscape of dendritic and somatic regions of neurons

定义神经元树突和体细胞区域的翻译景观

• 批准号: 8804069
• 负责人: Calvin Jan
• 金额: $ 11.46万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-19 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8804069
• 关键词: Address; Affect; Award; Axon; Bathing; Bicuculline; Biochemical; Biological Models; Biology; Biotin; Biotinylation; Brain; Calcium; Cataloging; Catalogs; Cells; Cellular Structures; Cellular biology; Collection; Cyclic AMP-Dependent Protein Kinases; Data Analyses; Dendrites; Electrophysiology (science); Engineering; Enzymes; FMRP; Faculty; Foundations; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Translation; Genome; Glutamates; Goals; Growth; High-Throughput Nucleotide Sequencing; Hippocampus (Brain); Housing; Hydrogen Peroxide; Individual; Institution; Intellectual functioning disability; Label; Learning; Ligase; Long-Term Depression; Long-Term Potentiation; Maps; Marine Biology; Mass Spectrum Analysis; Memory; Mental Depression; Mentors; Messenger RNA; Methods; Microscopy; Modeling; Molecular; Monitor; Mutate; Mutation; Neurobiology; Neurodevelopmental Disorder; Neurons; Neuropil; Neurosciences; Pathway interactions; Phase; Phenols; Play; Population; Post-Translational Protein Processing; Postdoctoral Fellow; Preparation; Protein Biosynthesis; Proteins; Proteome; Proteomics; Psyche structure; Radial; Regulation; Research; Research Personnel; Resolution; Ribosomes; Role; Signal Pathway; Signal Transduction; Slice; Specificity; Synapses; Synaptic plasticity; System; Systems Biology; Techniques; Technology; Tetrodotoxin; Time; Tissues; Training; Translating; Translation Process; Translations; Tuberous sclerosis protein complex; ascorbate peroxidase; autism spectrum disorder; base; calmodulin-dependent protein kinase II; career; data acquisition; deep sequencing; excitatory neuron; experience; flexibility; genome-wide; human FRAP1 protein; inhibitor/antagonist; innovation; interest; knowledge base; neuronal cell body; phenoxy radical; post-doctoral training; public health relevance; research study; response; tool; translational approach

DESCRIPTION (provided by applicant): The goal of this application is to map the translational landscape of excitatory neurons in response to synaptic activity. My interest in this problem is motivated by the observations that dynamic regulation of translation is crucial for both long term potentiation (LTP) and long term depression (LTD), the two forms of synaptic plasticity thought to underlie the cellular basis of learning and memory. Despite the importance of translational control in synaptic plasticity, only a handful of genes have been studied in this context. As a postdoctoral fellow in Jonathan Weissman's lab, I have developed an innovative approach for translational profiling with subcellular resolution. This technique is based on ribosome profiling, the deep sequencing of ribosome- protected mRNA fragments, invented in the Weissman lab. My immediate goal is to apply the technical and analytical toolkit that I have developed during my graduate and postdoctoral training to explore fundamental questions in neuroscience. The aims of this application are to 1) understand which mRNAs are translated locally in dendrites, and how translation is regulated in the dendritic and somatic compartments in response to elevated or silenced synaptic activity; 2) develop precise genetically encoded tools to facilitate dendritic ribosome profiling and dendritic proteomics; 3) explore the rapid translational changes that occur during LTD and determine which changes drive the electrophysiological response to activity. Having a genome-scale view of the translational response to synaptic activity should generate numerous hypotheses that I will continue to pursue in my independent research group. In order to achieve these aims, I seek interdisciplinary mentored training in neuroscience, gene regulation and cell biology. Towards this end, I have assembled a team of mentors with expertise in: systems biology and genome-scale data acquisition and analysis (Jonathan Weissman), neuroscience, synaptic plasticity and electrophysiology (Roger Nicoll), circuit analysis and activity-dependent signaling (Zachary Knight) and proteomics and genetic interaction analysis (Nevan Krogran). These mentors represent a broad and relevant collection scientific perspectives as well as career perspectives, as it includes both recently hired junior faculty and experienced tenured faculty. This K99/R00 award provides a protected training period in which I will expand my knowledge base in neuroscience, for example by taking the graduate neuroscience course NS201 at UCSF and the intensive Marine Biology Lab Neurobiology summer course. Such training will greatly facilitate my long-term research goals of understanding how neurons respond to synaptic activity with subcellular specificity and how activity-dependent gene expression changes are integrated into a spatially precise response. My long-term career objective is to pursue these research interests as a tenure-tracked principle investigator in an interdisciplinary biology department at an academic research institution.

描述（由申请人提供）：本申请的目的是映射兴奋性神经元的翻译局势，以响应突触活动。我对这个问题的兴趣是由观察到翻译的动态调节对于长期增强（LTP）和长期抑郁（LTD）至关重要，这两种突触可塑性被认为是学习和记忆的细胞基础。尽管在突触可塑性方面具有转化控制的重要性，但在这种情况下，仅研究了少数基因。作为乔纳森·韦斯曼（Jonathan Weissman）实验室的博士后研究员，我开发了一种具有亚细胞分辨率的转化分析的创新方法。该技术基于核糖体分析， 在魏斯曼实验室中发明的核糖体保护mRNA片段的深度测序。我的直接目标是应用我在研究生和博士后培训期间开发的技术和分析工具包来探索神经科学中的基本问题。该应用的目的是1）了解哪些mRNA在树突中局部翻译，以及如何在树突和体细胞室中调节翻译，以响应升高或沉默的突触活动； 2）开发精确的遗传编码工具，以促进树突状核糖体分析和树突状蛋白质组学； 3）探索LTD期间发生的快速翻译变化，并确定哪种变化促进了电生理对活动的反应。具有对突触活动的转化反应的基因组规模的观点，应产生许多假设，我将继续在我的独立研究小组中追求这些假设。为了实现这些目标，我寻求跨学科的指导神经科学，基因调节和细胞生物学培训。为此，我组建了一支具有专业知识的导师团队：系统生物学和基因组规模的数据获取和分析（Jonathan Weissman）（Jonathan Weissman），神经科学，突触可塑性和电生理学（Roger Nicoll）（Roger Nicoll），电路分析和活动依赖性信号传导（Zachary knight）和蛋白质组学和Genetic Interactionaltacticational（Nevenan Krogran）。这些导师代表着广泛而相关的科学观点以及职业观点，因为它包括最近聘用的初级教师和经验丰富的终身教师。这项K99/R00奖提供了一个受保护的培训期，我将扩大我在神经科学方面的知识基础，例如，在UCSF上参加研究生神经科学课程NS201和强化的海洋生物学实验室神经生物学夏季课程。这种训练将极大地促进我的长期研究目标，即了解神经元如何以亚细胞特异性对突触活动做出反应，以及如何将活动依赖性基因表达变化整合到空间上精确的响应中。我的长期职业目标是作为学术研究机构跨学科生物学系的终身制主要研究者追求这些研究兴趣。