Optogenetic imaging of protein activity in the synapse by using 2-photon fluorescence lifetime imaging microscopy. Optogenetics -Light-Sensing Proteins and Their Applications-.
Optogenetic imaging of protein activity in the synapse by using 2-photon fluorescence lifetime imaging microscopy. Optogenetics -Light-Sensing Proteins and Their Applications-.
复制标题
使用 2 光子荧光寿命成像显微镜对突触中的蛋白质活性进行光遗传学成像。
DOI：
10.1007/978-4-431-55516-2_12
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Murakoshi H.* and Shibata ACE.
中科院分区：
文献类型：
--
作者：
Murakoshi H;Shibata AC;Nakahata Y;and Nabekura J;Murakoshi H.* and Shibata ACE.
Elucidating the spatiotemporal dynamics of proteins in living cells, including protein–protein interactions and conformational changes, is essential for understanding the mechanisms of intracellular signal transduction and cellular functions such as synaptic plasticity, cell motility, and cell division. One of the best ways to visualize protein activity with high spatiotemporal resolution is to utilize optogenetic probes, such as green fluorescent protein, in combination with fluorescence resonance energy transfer (FRET) techniques, which enable us to measure the distance between donor and acceptor fluorescent proteins fused to the signaling proteins. Among the various FRET detection methods, 2-photon fluorescence lifetime imaging microscopy (2pFLIM) is the ideal method to monitor FRET in subcellular compartments of living cells located deep within tissues, such as in brain slices. This review introduces the principle of the 2pFLIM-FRET for monitoring intracellular protein activities and protein–protein interactions using two examples: detecting small GTPase activity and monitoring actin polymerization in dendrites and synapses of hippocampal neurons in brain slices.
