Molecular Substrates of Translation at the Synapse

描述（由申请人提供）：尽管对记忆存储中局部翻译的意义进行了数十年的研究，但对在特定突触处翻译的不同RNA的身份、其化学计量变化的时间窗口以及它们如何在任何系统中的突触处被利用知之甚少。RNA作为核糖核蛋白复合物通过分子马达主动转运到突触。在小鼠大脑中发现了40多种不同的RNA结合蛋白。我们已经开发了一种基因组学的方法来确定所有的RNA运输到突触的重点是积极运输的核糖核蛋白复合物。该策略涉及从感兴趣的组织中分离和表征免疫沉淀的分子马达复合物。我们建议通过分离和表征由驱动蛋白（一种微管依赖的分子马达，从细胞体移动到远端神经元过程）转运的蛋白复合物来识别和表征转运到海马和前额皮质（PFC）突触的编码和非编码RNA。该项目的长期目标是识别和表征突触翻译的分子底物及其在海马和PFC突触的长期记忆存储的启动和持续期间的时间调节，这两个关键的大脑区域参与长期记忆的存储和巩固。 公共卫生相关性：该项目以小鼠大脑的海马和前额叶皮层神经元为重点，将识别所有主动运输到突触的编码和非编码RNA。这些数据将用于识别和表征突触处的信号网络，这些信号网络在长期记忆的存储和巩固过程中受到局部翻译的调节。

DESCRIPTION (provided by applicant): Despite decades of research on the significance of local translation in memory storage, very little is known about the identity of the different RNAs translated at specific synapses, the temporal window of their stoichiometric changes, and how they are utilized at synapses in any system. RNAs are actively transported to synapses as ribo-nucleoprotein complexes by molecular motors. There are more than 40 different RNA binding proteins identified in a mouse brain. We have developed a genomics approach to identify all RNAs transported to synapses by focusing on the actively transported ribo-nucleoprotein complexes. This strategy involves isolation and characterization of immunoprecipitated molecular motor complexes from the tissue of interest. We propose to identify and characterize coding and noncoding RNAs transported to the hippocampal and prefrontal cortex (PFC) synapses by isolation and characterization of protein complexes transported by kinesin, a microtubule dependent molecular motor that moves from the cell-body to distal neuronal processes. The long-term goal of this project is to identify and characterize molecular substrates of synaptic translation and their temporal regulation during initiation and persistence of long-term memory storage at the hippocampal and PFC synapses, the two critical brain regions involved in storage and consolidation of long-term memories. PUBLIC HEALTH RELEVANCE: Focusing on the hippocampal and prefrontal cortex neurons of the mouse brain, this project will identify all the coding and noncoding RNAs that are actively transported to synapses. This data will be used to identify and characterize signaling networks at the synapse that are regulated by local translation during storage and consolidation of long-term memories.