Wehrle-Haller Bernhard
Department of Cellular Physiology and Metabolism, Centre Medical Universitaire, University of Geneva, Geneva, Switzerland.
Methods Mol Biol. 2007;370:173-202. doi: 10.1007/978-1-59745-353-0_13.
Cell migration is a complex cellular behavior that involves the controlled reorganization of the link between the actin cytoskeleton and the extracellular matrix. This mechanical connection is provided by transmembrane receptors of the integrin family. Integrins are heterodimeric receptors that undergo an allosteric switch when activated by external or intracellular signals, providing binding sites for ligands of the extracellular matrix and actin-associated cytoplasmic adapter proteins. Techniques such as fluorescence recovery after photobleaching (FRAP) are used to analyze the remodeling of green fluorescent protein (GFP)-tagged integrin receptors within focal adhesions, demonstrating that the dynamics of the remodeling of integrins in substrate adhesion sites is carefully regulated by extracellular ligands, cytoskeletal adapter proteins, and the actin cytoskeleton. FRAP analysis of GFP-tagged integrins is a tool that allows one to detect and dissect the internal dynamics of apparently immobile focal adhesions, allowing one to perceive the hierarchies and mechanisms of focal adhesion formation and dispersion.
细胞迁移是一种复杂的细胞行为，涉及肌动蛋白细胞骨架与细胞外基质之间连接的受控重组。这种机械连接由整合素家族的跨膜受体提供。整合素是异二聚体受体，当被外部或细胞内信号激活时会发生变构转换，为细胞外基质的配体和肌动蛋白相关的细胞质衔接蛋白提供结合位点。诸如光漂白后荧光恢复（FRAP）等技术用于分析粘着斑内绿色荧光蛋白（GFP）标记的整合素受体的重塑，表明底物粘附位点中整合素重塑的动力学受到细胞外配体、细胞骨架衔接蛋白和肌动蛋白细胞骨架的精细调节。对GFP标记的整合素进行FRAP分析是一种工具，可用于检测和剖析看似固定的粘着斑的内部动力学，使人们能够了解粘着斑形成和分散的层次结构及机制。
