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3D Coupling of Fibronectin Fibril Arrangement with Topology of Ventral Plasma Membrane

机译:纤连蛋白原纤维排列与腹膜质膜拓扑的3D耦合

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Interaction of integrins with extracellular matrices is essential for cell adhesion to substrata. Ventral surfaces of fibroblasts adhering to flat substrata are not flat but have uneven 3D topology. However, spatial relationship between the topology of the ventral cell surface and arrangement of extracellular matrix fibrils remains unclear. Here, we report a novel and simple method based on total internal reflection fluorescence microscopy to quantify the distance between the ventral plasma membrane and the glass substratum. We observe that the distance varies from ?25 nm at focal adhesions to 40–50 nm at close contacts and ?80 nm in other regions. Furthermore, by applying this novel method, we show that fibronectin fibrils are also separated from the substratum in regions where the ventral cell surface-substratum distance is ?80 nm. Our results reveal that fibronectin fibrils are not simply adsorbed to the glass substratum but follow the ventral cell surface topology.
机译:整联蛋白与细胞外基质的相互作用对于细胞粘附至基质至关重要。附着在平坦基底上的成纤维细胞的腹表面不平坦,但具有不均匀的3D拓扑。然而,腹侧细胞表面的拓扑结构和细胞外基质原纤维的排列之间的空间关系仍然不清楚。在这里,我们报告了一种基于全内反射荧光显微镜的新颖而简单的方法,以量化腹膜质膜和玻璃基质之间的距离。我们观察到,距离从粘着斑处的<?25 nm到紧密接触处的40–50 nm,在其他区域的>?80 nm处变化。此外,通过应用这种新颖的方法,我们表明纤连蛋白原纤维在腹侧细胞表面-基质距离> 80nm的区域中也与基质分离。我们的研究结果表明纤连蛋白原纤维不仅被简单地吸附到玻璃基底上,而且遵循腹侧细胞表面的拓扑结构。

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