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）：本申请的目标是绘制兴奋性神经元响应突触活动的翻译图谱。我对这个问题的兴趣源于这样的观察：翻译的动态调节对于长期增强（LTP）和长期抑制（LTD）都至关重要，这两种形式的突触可塑性被认为是学习和记忆的细胞基础。尽管翻译控制在突触可塑性中很重要，但在这方面只研究了少数基因。作为 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翻译分析的创新方法。该技术基于核糖体分析， 对核糖体保护的 mRNA 片段进行深度测序，由 Weissman 实验室发明。我的近期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的基本问题。该应用的目的是 1) 了解哪些 mRNA 在树突中进行局部翻译，以及如何在树突和体细胞区室中调节翻译以响应突触活动的升高或沉默； 2）开发精确的基因编码工具以促进树突核糖体分析和树突蛋白质组学； 3) 探索LTD期间发生的快速转化变化，并确定哪些变化驱动对活动的电生理反应。对突触活动的翻译反应有一个基因组规模的看法应该会产生许多假设，我将在我的独立研究小组中继续追求这些假设。为了实现这些目标，我寻求神经科学、基因调控和细胞生物学方面的跨学科指导培训。为此，我组建了一支具有以下专业知识的导师团队：系统生物学和基因组规模的数据采集和分析（Jonathan Weissman），神经科学，突触可塑性和电生理学（Roger Nicoll），电路分析和活动依赖性信号传导（Zachary） Knight）以及蛋白质组学和遗传相互作用分析（Nevan Krogran）。这些导师代表了广泛且相关的科学观点和职业观点，因为其中包括最近聘用的初级教师和经验丰富的终身教员。这个 K99/R00 奖项提供了一个受保护的培训期，在此期间我将扩展我在神经科学方面的知识基础，例如通过参加 UCSF 的研究生神经科学课程 NS201 和强化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，以及活动依赖性基因表达变化如何整合到空间精确的反应中。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教授首席研究员来追求这些研究兴趣。