力传导中亚细胞水平的分子FRET成像（英文）

FRET imaging of molecular hierarchy at subcellular levels in mechanotransduction

体内细胞受到含有化学和力学因素的生理和病理生理的刺激,故研究这些因素在细胞和器官水平如何调节功能就尤为重要。有关细胞和器官对化学因素的反应已开展诸多研究,而力学因素的影响却鲜有报道。近年来,荧光蛋白和显微镜技术的发展已成为阐明力传导过程的有用工具,先进的信号活细胞成像技术促进了力学生物学中分子机制的时空因素研究。本文综述荧光蛋白的基本知识以及其在生物学研究中的应用,特别讨论了以荧光共振能量迁移(fluorescence proteins and microscopy,FRET)技术为基础的生物传感器的发展和特征。基因编码的FRET生物传感器能够实现分子时空活动的成像和定量,使得活细胞中生物化学信号在力学刺激下的反应和传导可视化。同时,本文重点阐述分子水平力学刺激下的活细胞信号传导。

Cells in the body are exposed to physiological and pathophysiological stimuli that encompass both chemical and mechanical factors. It is important to understand how these factors modulate functions at cellular and organ levels. Compared to the large amount of information on cellular or organ responses to chemical factors, there is a paucity of knowledge on the effects of mechanical factors. Recent advances of fluorescence proteins and microscopy make it a very useful tool for elucidating the mechanotransduction processes; the state-of- the-art technologies for live-cell imaging of signaling is particularly valuable for investigating the spatia and temporal aspects of molecular mechanisms in mechanobiology. This review will cover the basic knowledge of fluores- cence proteins and their application for biological research. In particular, the development and characterization of biosensors based on fluorescent resonance energy transfer （FRET） will be discussed. Genetically encoded FRET biosensors, which allows the imaging and quantification of tempo-spatial activation of molecules, will be introduced to demonstrate how the initiation and transmission of biochemical signals in response to local mechanical stimulation can be visualized in live cells. Specific emphasis will be on the elucidation of molecule hierarchy of signaling transduction in live cells upon the mechanical stimulation.