Techniques for studying protein trafficking and molecular motors in neurons.
Techniques for studying protein trafficking and molecular motors in neurons.
复制标题
研究神经元中蛋白质运输和分子马达的技术。
DOI：
10.1002/cm.21274
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Arnold,DonB
中科院分区：
文献类型：
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作者：
Feng,Shanxi;Arnold,DonB
本文综述了促进蛋白质运输可视化的技术。在20世纪90年代中期，GFP的克隆使荧光标记蛋白在细胞中表达，然后实时可视化。这一进展使我们第一次得以窥见细胞内分配蛋白质的复杂系统。然而，它很快变得明显，外源表达的GFP标记蛋白的时间推移序列可能难以解释。其原因包括来自移动蛋白的相对较低的信号和来自静止蛋白和其他来源的高背景速率，以及难以识别特定囊泡载体的起源和目的地。在这篇综述中，回顾了不同程度上克服这些问题的一系列技术，并讨论了它们对蛋白质运输的见解。注意力将集中在神经元上，因为它们是高度极化的，因此，它们的传输系统往往易于研究。©2016 Wiley期刊公司
This review focused on techniques that facilitated the visualization of protein trafficking. In the mid‐1990s the cloning of GFP allowed fluorescently tagged proteins to be expressed in cells and then visualized in real time. This advance allowed a glimpse, for the first time, of the complex system within cells for distributing proteins. It quickly became apparent, however, that time‐lapse sequences of exogenously expressed GFP‐labeled proteins can be difficult to interpret. Reasons for this include the relatively low signal that comes from moving proteins and high background rates from stationary proteins and other sources, as well as the difficulty of identifying the origins and destinations of specific vesicular carriers. In this review a range of techniques that have overcome these issues to varying degrees was reviewed and the insights into protein trafficking that they have enabled were discussed. Concentration will be on neurons, as they are highly polarized and, thus, their trafficking systems tend to be accessible for study. © 2016 Wiley Periodicals, Inc.