Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes.
Transport and localization of exogenous myelin basic protein mRNA microinjected into oligodendrocytes.
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DOI：
10.1083/jcb.123.2.431
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发表时间：
1993-10
期刊：
影响因子：
--
通讯作者：
Carson JH
中科院分区：
文献类型：
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作者：
Ainger K;Avossa D;Morgan F;Hill SJ;Barry C;Barbarese E;Carson JH
我们已经研究了运输和定位的MBP mRNA在少突胶质细胞培养的标记的mRNA注射到活细胞和分析的细胞内分布的注射RNA的共聚焦显微镜。注射的mRNA最初似乎分散在核周体中。在几分钟内，RNA形成颗粒，在MBP mRNA的情况下，这些颗粒沿着突起向下运输到细胞的外围，在那里分布再次分散。原位杂交结果显示，少突胶质细胞内的内源性MBP mRNA也以颗粒形式存在于细胞核周围和突起中，并弥散分布于细胞膜上。用非离子去污剂提取时，颗粒未释放，表明它们与细胞骨架基质相关。三维可视化显示，MBP mRNA颗粒经常排列在轨道沿着微管穿越细胞质和过程。观察到几种不同的颗粒运动模式。过程中的颗粒以约0.2微米/秒的速度进行持续的定向运动。分支点处的颗粒经历平均位移为0.1微米/秒的振荡运动。细胞周围的颗粒随机循环，平均位移约为1微米/秒。结果进行了讨论，在一个多步骤的少突胶质细胞中的MBP mRNA的运输和本地化的途径。这项工作代表了活细胞中mRNA的细胞内运动的第一个特征，以及RNA颗粒在细胞中mRNA的运输和定位中的作用的第一个描述。
We have studied transport and localization of MBP mRNA in oligodendrocytes in culture by microinjecting labeled mRNA into living cells and analyzing the intracellular distribution of the injected RNA by confocal microscopy. Injected mRNA initially appears dispersed in the perikaryon. Within minutes, the RNA forms granules which, in the case of MBP mRNA, are transported down the processes to the periphery of the cell where the distribution again becomes dispersed. In situ hybridization shows that endogenous MBP mRNA in oligodendrocytes also appears as granules in the perikaryon and processes and dispersed in the peripheral membranes. The granules are not released by extraction with non-ionic detergent, indicating that they are associated with the cytoskeletal matrix. Three dimensional visualization indicates that MBP mRNA granules are often aligned in tracks along microtubules traversing the cytoplasm and processes. Several distinct patterns of granule movement are observed. Granules in the processes undergo sustained directional movement with a velocity of approximately 0.2 micron/s. Granules at branch points undergo oscillatory motion with a mean displacement of 0.1 micron/s. Granules in the periphery of the cell circulate randomly with a mean displacement of approximately 1 micron/s. The results are discussed in terms of a multi-step pathway for transport and localization of MBP mRNA in oligodendrocytes. This work represents the first characterization of intracellular movement of mRNA in living cells, and the first description of the role of RNA granules in transport and localization of mRNA in cells.