Defining the translational landscape of dendritic and somatic regions of neurons

描述（由申请人提供）：本申请的目标是绘制兴奋性神经元对突触活动反应的翻译景观。我之所以对这个问题感兴趣，是因为观察到翻译的动态调节对长期增强（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，但只有少数基因在这方面得到了研究。作为Jonathan Weissman实验室的博士后研究员，我开发了一种创新的亚细胞分辨率翻译分析方法。这项技术是基于核糖体分析，

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.