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      Isotropic actomyosin dynamics promote organization of the apical cell cortex in epithelial cells

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          Abstract

          Apical membrane organization of nonconfluent epithelial cells is driven by a dynamic network of actin and myosin II filaments.

          Abstract

          Although cortical actin plays an important role in cellular mechanics and morphogenesis, there is surprisingly little information on cortex organization at the apical surface of cells. In this paper, we characterize organization and dynamics of microvilli (MV) and a previously unappreciated actomyosin network at the apical surface of Madin–Darby canine kidney cells. In contrast to short and static MV in confluent cells, the apical surfaces of nonconfluent epithelial cells (ECs) form highly dynamic protrusions, which are often oriented along the plane of the membrane. These dynamic MV exhibit complex and spatially correlated reorganization, which is dependent on myosin II activity. Surprisingly, myosin II is organized into an extensive network of filaments spanning the entire apical membrane in nonconfluent ECs. Dynamic MV, myosin filaments, and their associated actin filaments form an interconnected, prestressed network. Interestingly, this network regulates lateral mobility of apical membrane probes such as integrins or epidermal growth factor receptors, suggesting that coordinated actomyosin dynamics contributes to apical cell membrane organization.

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          Most cited references54

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          Epithelial-mesenchymal transitions in development and disease.

          The epithelial to mesenchymal transition (EMT) plays crucial roles in the formation of the body plan and in the differentiation of multiple tissues and organs. EMT also contributes to tissue repair, but it can adversely cause organ fibrosis and promote carcinoma progression through a variety of mechanisms. EMT endows cells with migratory and invasive properties, induces stem cell properties, prevents apoptosis and senescence, and contributes to immunosuppression. Thus, the mesenchymal state is associated with the capacity of cells to migrate to distant organs and maintain stemness, allowing their subsequent differentiation into multiple cell types during development and the initiation of metastasis.
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            Actin, a central player in cell shape and movement.

            The protein actin forms filaments that provide cells with mechanical support and driving forces for movement. Actin contributes to biological processes such as sensing environmental forces, internalizing membrane vesicles, moving over surfaces, and dividing the cell in two. These cellular activities are complex; they depend on interactions of actin monomers and filaments with numerous other proteins. Here, we present a summary of the key questions in the field and suggest how those questions might be answered. Understanding actin-based biological phenomena will depend on identifying the participating molecules and defining their molecular mechanisms. Comparisons of quantitative measurements of reactions in live cells with computer simulations of mathematical models will also help generate meaningful insights.
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              Feature point tracking and trajectory analysis for video imaging in cell biology.

              This paper presents a computationally efficient, two-dimensional, feature point tracking algorithm for the automated detection and quantitative analysis of particle trajectories as recorded by video imaging in cell biology. The tracking process requires no a priori mathematical modeling of the motion, it is self-initializing, it discriminates spurious detections, and it can handle temporary occlusion as well as particle appearance and disappearance from the image region. The efficiency of the algorithm is validated on synthetic video data where it is compared to existing methods and its accuracy and precision are assessed for a wide range of signal-to-noise ratios. The algorithm is well suited for video imaging in cell biology relying on low-intensity fluorescence microscopy. Its applicability is demonstrated in three case studies involving transport of low-density lipoproteins in endosomes, motion of fluorescently labeled Adenovirus-2 particles along microtubules, and tracking of quantum dots on the plasma membrane of live cells. The present automated tracking process enables the quantification of dispersive processes in cell biology using techniques such as moment scaling spectra.
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                Author and article information

                Journal
                J Cell Biol
                J. Cell Biol
                jcb
                jcb
                The Journal of Cell Biology
                The Rockefeller University Press
                0021-9525
                1540-8140
                13 October 2014
                : 207
                : 1
                : 107-121
                Affiliations
                [1 ]Cellular Dynamics and Cell Patterning and [2 ]Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany
                [3 ]Institute of Cell Dynamics and Imaging and [4 ]Cells-In-Motion Cluster of Excellence (EXC1003–CiM), University of Münster, 48149 Münster, Germany
                [5 ]Laboratory for Computational Biology & Biophysics, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139
                [6 ]Max Planck Institute of Molecular Cell Biology and Genetics, 01307 Dresden, Germany
                [7 ]Division of Molecular Biology, Ruđer Bošković Institute, 10000 Zagreb, Croatia
                Author notes
                Correspondence to Roland Wedlich-Soldner: wedlich@ 123456uni-muenster.de

                C. Klingner and A.V. Cherian contributed equally to this paper.

                Article
                201402037
                10.1083/jcb.201402037
                4195824
                25313407
                f7c7109d-bf41-48b1-b42d-d7a728292ce4
                © 2014 Klingner et al.

                This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

                History
                : 7 February 2014
                : 22 August 2014
                Categories
                Research Articles
                Article

                Cell biology
                Cell biology

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