Studying early stages of fibronectin fibrillogenesis in living cells by atomic force microscopy скачать в хорошем качестве

Studying early stages of fibronectin fibrillogenesis in living cells by atomic force microscopy

Studying early stages of fibronectin fibrillogenesis in living cells by atomic force microscopy. Tetyana Gudzenko and Clemens M. Franz (2015), Molecular Biology of the Cell http://dx.doi.org/10.1091/mbc.E14-05-... Fibronectin (FN) is an extracellular matrix protein that can be assembled by cells into large fibrillar networks, but the dynamics of FN remodeling and the transition through intermediate fibrillar stages are still incompletely understood. Here we have used a combination of fluorescence microscopy and time-lapse atomic force microscopy (AFM) to visualize initial stages of FN fibrillogenesis in living fibroblasts at high resolution. Initial FN nanofibrils form within 5 min of cell-matrix contact and subsequently extend at a rate of 0.25 μm/min at sites of cell membrane retraction. FN nanofibrils display a complex linear array of globular features spaced at varying distances, indicating the coexistence of different conformational states within the fibril. In some cases initial fibrils extended in discrete increments of ∼800 nm during a series of cyclical membrane retractions, indicating a stepwise fibrillar extension mechanism. In presence of Mn2+, a known activator of integrin adhesion to FN, fibrillogenesis was accelerated almost threefold to 0.68 μm/min and fibrillar dimensions were increased, underlining the importance of integrin activation for early FN fibrillogenesis. FN fibrillogenesis visualized by time-lapse AFM thus provides new structural and mechanistic insight into initial steps of cell-driven FN fibrillogenesis. Good channel:    / dlium   Subscribe, like and comment. Good website: https://www.dlium.com Bookmark, subscribe and comment.