González-González Inmaculada M, Jaskolski Frédéric, Goldberg Yves, Ashby Michael C, Henley Jeremy M
MRC Centre for Synaptic Plasticity, School of Biochemistry, Medical Sciences Building, University of Bristol, Bristol, United Kingdom.
Methods Enzymol. 2012;504:127-46. doi: 10.1016/B978-0-12-391857-4.00006-9.
The use of genetically encoded fluorescent tags such as green fluorescent protein (GFP) as reporters to monitor processes in living cells has transformed cell biology. One major application for these tools has been to analyze protein dynamics in neurons. In particular, fluorescence recovery after photobleach (FRAP) of surface expressed fluorophore-tagged proteins has been instrumental to addressing outstanding questions about how neurons orchestrate the synaptic delivery of proteins. Here, we provide an overview of the methodology, equipment, and analysis required to perform, analyze, and interpret these experiments.
使用诸如绿色荧光蛋白（GFP）等基因编码荧光标签作为报告分子来监测活细胞中的过程，已经改变了细胞生物学。这些工具的一个主要应用是分析神经元中的蛋白质动态。特别是，对表面表达的荧光团标记蛋白进行光漂白后的荧光恢复（FRAP），对于解决有关神经元如何协调蛋白质的突触传递的突出问题起到了重要作用。在这里，我们概述了进行、分析和解释这些实验所需的方法、设备和分析。
