Molecular Substrates of Translation at the Synapse

描述(申请人提供)：尽管对局部翻译在记忆存储中的重要性进行了数十年的研究，但人们对在特定突触翻译的不同RNA的身份、它们化学计量学变化的时间窗口以及它们如何在任何系统中的突触中被利用知之甚少。RNA通过分子马达以核糖核蛋白复合体的形式主动转运到突触。在小鼠的大脑中发现了40多种不同的RNA结合蛋白。我们开发了一种基因组学方法，通过专注于活跃运输的核糖核蛋白复合体来识别所有运输到突触的RNA。这一策略包括从感兴趣的组织中分离和鉴定免疫沉淀的分子马达复合体。我们建议通过分离和表征由Kinesin运输的蛋白质复合体来识别和表征运输到海马和前额叶皮质(PFC)突触的编码和非编码RNA。Kinesin是一种微管依赖的分子马达，从细胞体移动到远端神经元突起。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突触长期记忆存储的启动和持续期间的时间调节，这两个大脑区域参与长期记忆的存储和巩固。 与公共卫生相关：该项目将聚焦于小鼠大脑的海马区和前额叶皮质神经元，识别所有活跃地运输到突触的编码和非编码RNA。这些数据将被用来识别和描述突触上的信号网络，这些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.