Localized mRNAs in Synaptic Plasticity

项目摘要 mRNA定位和调节翻译在轴突导向和突触可塑性中起着重要作用。通过 在神经元内的空间限制基因表达，局部蛋白质合成允许生长锥和突触 自主调节它们的结构和功能。本项目的长期目标是了解 学习相关神经元可塑性和突触过程中局部翻译的细胞生物学和功能 阵我们的实验旨在解决以下四个问题：1）存在哪些mRNA 在神经过程中？2)mRNA如何定位于神经元内？3)如何翻译本地化的成绩单 在突触处受到调节4）神经元中特定的局部翻译蛋白的功能是什么？我们 在一个特征良好的、实验上易于处理的突触发生模型中解决这些问题， 学习相关的突触可塑性：培养的感觉运动神经元。我们的战略包括使用 微阵列和高通量测序技术来鉴定存在于神经元中的mRNA， 神经元刺激后的过程，以及细胞生物学方法来可视化mRNA 在活神经元中的定位和调节翻译以及RNA干扰方法来研究功能 在突触形成和突触可塑性过程中特定的局部mRNA的表达。我们提出的结果 实验将提供深入了解mRNA定位在神经元内的细胞生物学和功能， 特定的本地化转录本。

Project Summary mRNA localization and regulated translation play central roles in axon guidance and synaptic plasticity. By spatially restricting gene expression within neurons, local protein synthesis allows growth cones and synapses to autonomously regulate their structure and function. The long-term goals of this project are to understand the cell biology and function of local translation during learning-related neuronal plasticity and during synapse formation. Our experiments are designed to address the following four questions: 1) what mRNAs are present in neuronal processes? 2) how do mRNAs localize within neurons? 3) how is translation of localized transcripts regulated at the synapse? and 4) what is the function of specific locally translated proteins in neurons? We address these questions in a well-characterized, experimentally tractable model of synaptogenesis and learning-related synaptic plasticity: cultured Aplysia sensory-motor neurons. Our strategy involves the use of microarray and high throughput sequencing technologies to identify mRNAs that are present in neuronal processes following neuronal stimulation, together with cell biological approaches to visualize mRNA localization and regulated translation in living neurons and RNA interference approaches to study the function of specific localized mRNAs during synapse formation and synaptic plasticity. The results of our proposed experiments will provide insight into the cell biology of mRNA localization within neurons and into the function of specific localized transcripts.