Glutamate-induced RNA localization and translation in neurons
Glutamate-induced RNA localization and translation in neurons
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DOI：
10.1073/pnas.1614267113
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发表时间：
2016-11-01
影响因子：
11.1
通讯作者：
Singer, Robert H.
中科院分区：
文献类型：
--
作者：
Yoon, Young J.;Wu, Bin;Singer, Robert H.
mRNA的定位是蛋白质合成在离散的细胞内区室中发生所必需的。神经元是研究mRNA运输精确性的理想系统，因为它们的极化结构和突触特异性靶向的需要。为了研究这种定位，我们得出了一个定量和分析的方法。树突棘刺激谷氨酸uncaging在衍射有限的斑点，并在空间和时间的定位测量单个β-肌动蛋白mRNA。定位需要NMDA受体活性，动态肌动蛋白细胞骨架，和transacting RNA结合蛋白，邮编结合蛋白1（ZBP 1）。使用Halo-actin报告基因评估mRNA将新合成的蛋白质引导至定位位点的能力，从而同时检测RNA和蛋白质。新合成的Halo-肌动蛋白在刺激位点富集，需要NMDA受体活性，并且优先定位在棘的外周。这项工作表明，突触活动可以诱导mRNA定位和β-肌动蛋白的局部翻译，其中新的肌动蛋白参与稳定树突棘中扩展的突触。
Localization of mRNA is required for protein synthesis to occur within discrete intracellular compartments. Neurons represent an ideal system for studying the precision of mRNA trafficking because of their polarized structure and the need for synapse-specific targeting. To investigate this targeting, we derived a quantitative and analytical approach. Dendritic spines were stimulated by glutamate uncaging at a diffraction-limited spot, and the localization of single beta-actin mRNAs was measured in space and time. Localization required NMDA receptor activity, a dynamic actin cytoskeleton, and the transacting RNA-binding protein, Zipcode-binding protein 1 (ZBP1). The ability of the mRNA to direct newly synthesized proteins to the site of localization was evaluated using a Halo-actin reporter so that RNA and protein were detected simultaneously. Newly synthesized Halo-actin was enriched at the site of stimulation, required NMDA receptor activity, and localized preferentially at the periphery of spines. This work demonstrates that synaptic activity can induce mRNA localization and local translation of beta-actin where the new actin participates in stabilizing the expanding synapse in dendritic spines.