Identification of axon-enriched microRNAs localized to growth cones of cortical neurons.
Identification of axon-enriched microRNAs localized to growth cones of cortical neurons.
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DOI：
10.1002/dneu.22113
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发表时间：
2014-03
影响因子：
3
通讯作者：
Bassell, Gary J.
中科院分区：
文献类型：
--
作者：
Sasaki, Yukio;Gross, Christina;Xing, Lei;Goshima, Yoshio;Bassell, Gary J.
越来越多的证据表明，轴突和生长锥中的局部mRNA通过局部翻译在轴突延伸和寻路中发挥重要作用。一些研究已经揭示了轴突中存在微小RNA（miRNAs），其可以控制轴突发育期间的局部蛋白质合成。然而，到目前为止，还没有尝试筛选轴突富集的miRNA并验证它们可能定位于来自中枢神经系统神经元的发育轴突的生长锥。在这项研究中，使用“神经元球”培养方法检查了miRNAs在皮层神经元轴突和生长锥中的定位，该方法适合于以高产量和纯度制备轴突miRNAs。通过定量实时RT-PCR分析从小鼠皮层神经元的神经元球培养物制备的轴突miRNA。在分析的375个miRNAs中，105个miRNAs在轴突中检测到，6个miRNAs与细胞体组分相比在轴突组分中显著富集。荧光原位杂交显示，两个轴突富集的miRNA，miR-181 a-1* 和miR-532，定位为不同的颗粒在远端轴突和生长锥。这些miRNA与RNA诱导的沉默复合物的结合进一步支持了它们调节脑中mRNA水平或翻译的功能。这些结果表明，特定的miRNAs定位于远端轴突和生长锥的机制，在那里它们可能参与局部mRNA的调节。这些发现为轴突miRNAs的存在提供了新的见解，并促使进一步分析其在轴突指导下的局部蛋白质合成中的功能。
There is increasing evidence that localized mRNAs in axons and growth cones play an important role in axon extension and pathfinding via local translation. A few studies have revealed the presence of microRNAs (miRNAs) in axons, which may control local protein synthesis during axon development. However, so far, there has been no attempt to screen for axon-enriched miRNAs and to validate their possible localization to growth cones of developing axons from neurons of the central nervous system. In this study, the localization of miRNAs in axons and growth cones in cortical neurons was examined using a “neuron ball” culture method that is suitable to prepare axonal miRNAs with high yield and purity. Axonal miRNAs prepared from the neuron ball cultures of mouse cortical neurons were analyzed by quantitative real-time RT-PCR. Among 375 miRNAs that were analyzed, 105 miRNAs were detected in axons, and six miRNAs were significantly enriched in axonal fractions when compared with cell body fractions. Fluorescence in situ hybridization revealed that two axon-enriched miRNAs, miR-181a-1* and miR-532, localized as distinct granules in distal axons and growth cones. The association of these miRNAs with the RNA-induced silencing complex further supported their function to regulate mRNA levels or translation in the brain. These results suggest a mechanism to localize specific miRNAs to distal axons and growth cones, where they could be involved in local mRNA regulation. These findings provide new insight into the presence of axonal miRNAs and motivate further analysis of their function in local protein synthesis underlying axon guidance.
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未损伤和再生的哺乳动物皮质轴突中的轴突 mRNA。
DOI：
10.1523/jneurosci.6130-08.2009
发表时间：
2009-04-15
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
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作者：
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影响因子：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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DOI：
10.1146/annurev.cellbio.20.010403.111746
发表时间：
2004
影响因子：
11.3
作者：
Piper M;Holt C
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