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综述:整合素研究中的成像技术

Overview: imaging in the study of integrins.

作者信息

Carman Christopher V

机构信息

Center for Vascular Biology Research, Division of Molecular and Vascular Medicine, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.

出版信息

Methods Mol Biol. 2012;757:159-89. doi: 10.1007/978-1-61779-166-6_12.

Abstract

Integrins play critical adhesion and signaling roles during development, wound healing, immunity, and cancer. Central to their function is a unique ability to dynamically modulate their adhesiveness and signaling properties through changes in conformation, both homo- and heterotypic protein-protein interactions and cellular distribution. Genetic, biochemical and structural studies have been instrumental in uncovering overall functions, describing ligand and regulatory protein interactions and elucidating the molecular architecture of integrins. However, such approaches alone are inadequate to describe how dynamic integrin behaviors are orchestrated in intact cells. To fill this void, a wide array of distinct light microscopy (largely fluorescence-based) imaging approaches have been developed and employed. Various microscopy technologies, including wide-field, optical sectioning (laser-scanning confocal, spinning-disk confocal, and multiphoton), TIRF and range of novel "Super-Resolution" techniques have been used in combination with diverse imaging modalities (such as IRM, FRET, FRAP, CALI, and fluorescence speckle imaging) to address distinct aspects of integrin function and regulation. This chapter provides an overview of these imaging approaches and how they have advanced our understanding of integrins.

摘要

整合素在发育、伤口愈合、免疫和癌症过程中发挥着关键的黏附与信号传导作用。其功能的核心在于通过构象变化、同型和异型蛋白质-蛋白质相互作用以及细胞分布,动态调节其黏附性和信号特性的独特能力。遗传、生化和结构研究有助于揭示整合素的整体功能、描述配体和调节蛋白相互作用以及阐明整合素的分子结构。然而,仅靠这些方法不足以描述完整细胞中整合素动态行为的调控机制。为填补这一空白,人们开发并采用了一系列不同的光学显微镜(主要基于荧光)成像方法。各种显微镜技术,包括宽场显微镜、光学切片显微镜(激光扫描共聚焦显微镜、转盘共聚焦显微镜和多光子显微镜)、全内反射荧光显微镜以及一系列新型“超分辨率”技术,已与多种成像模式(如干涉反射显微镜、荧光共振能量转移、荧光漂白恢复、化学诱导荧光淬灭和荧光斑点成像)结合使用,以研究整合素功能和调控的不同方面。本章概述了这些成像方法以及它们如何推动我们对整合素的理解。

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