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使用荧光显微镜进行单细胞成像翻译。

Imaging translation in single cells using fluorescent microscopy.

机构信息

Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.

出版信息

Cold Spring Harb Perspect Biol. 2012 Nov 1;4(11):a012310. doi: 10.1101/cshperspect.a012310.

Abstract

The regulation of translation provides a mechanism to control not only the abundance of proteins, but also the precise time and subcellular location that they are synthesized. Much of what is known concerning the molecular basis for translational control has been gleaned from experiments (e.g., luciferase assays and polysome analysis) that measure average changes in the protein synthesis of a population of cells, however, mechanistic insights can be obscured in ensemble measurements. The development of fluorescent microscopy techniques and reagents has allowed translation to be studied within its cellular context. Here we highlight recent methodologies that can be used to study global changes in protein synthesis or regulation of specific mRNAs in single cells. Imaging of translation has provided direct evidence for local translation of mRNAs at synapses in neurons and will become an important tool for studying translational control.

摘要

翻译的调控不仅提供了一种控制蛋白质丰度的机制,还提供了一种控制蛋白质合成的精确时间和亚细胞位置的机制。关于翻译调控的分子基础的大部分知识都是从实验中获得的(例如,荧光素酶测定和多核糖体分析),这些实验测量了细胞群体中蛋白质合成的平均变化,然而,在整体测量中,机制上的见解可能会被掩盖。荧光显微镜技术和试剂的发展使得可以在细胞环境中研究翻译。在这里,我们重点介绍了一些可以用于研究单个细胞中蛋白质合成的整体变化或特定 mRNA 调控的最新方法。翻译的成像为神经元突触中 mRNA 的局部翻译提供了直接证据,并将成为研究翻译调控的重要工具。

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