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      Thrombin-induced cytoskeleton dynamics in spread human platelets observed with fast scanning ion conductance microscopy

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          Abstract

          Platelets are small anucleate blood cells involved in haemostasis. Platelet activation, caused by agonists such as thrombin or by contact with the extracellular matrix, leads to platelet adhesion, aggregation, and coagulation. Activated platelets undergo shape changes, adhere, and spread at the site of injury to form a blood clot. We investigated the morphology and morphological dynamics of human platelets after complete spreading using fast scanning ion conductance microscopy (SICM). In contrast to unstimulated platelets, thrombin-stimulated platelets showed increased morphological activity after spreading and exhibited dynamic morphological changes in the form of wave-like movements of the lamellipodium and dynamic protrusions on the platelet body. The increase in morphological activity was dependent on thrombin concentration. No increase in activity was observed following exposure to other activation agonists or during contact-induced activation. Inhibition of actin polymerization and inhibition of dynein significantly decreased the activity of thrombin-stimulated platelets. Our data suggest that these morphological dynamics after spreading are thrombin-specific and might play a role in coagulation and blood clot formation.

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          The restaurant at the end of the random walk: recent developments in the description of anomalous transport by fractional dynamics

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            Small-molecule inhibitors of the AAA+ ATPase motor cytoplasmic dynein

            The conversion of chemical energy into mechanical force by AAA+ (ATPases associated with diverse cellular activities) ATPases is integral to cellular processes, including DNA replication, protein unfolding, cargo transport, and membrane fusion 1 . The AAA+ ATPase motor cytoplasmic dynein regulates ciliary trafficking 2 , mitotic spindle formation 3 , and organelle transport 4 , and dissecting its precise functions has been challenging due to its rapid timescale of action and the lack of cell-permeable, chemical modulators. Here we describe the discovery of ciliobrevins, the first specific small-molecule antagonists of cytoplasmic dynein. Ciliobrevins perturb protein trafficking within the primary cilium, leading to their malformation and Hedgehog signaling blockade. Ciliobrevins also prevent spindle pole focusing, kinetochore-microtubule attachment, melanosome aggregation, and peroxisome motility in cultured cells. We further demonstrate the ability of ciliobrevins to block dynein-dependent microtubule gliding and ATPase activity in vitro. Ciliobrevins therefore will be useful reagents for studying cellular processes that require this microtubule motor and may guide the development of additional AAA+ ATPase superfamily inhibitors.
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              Biomedical Image Processing

              Sternberg (1983)
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                Author and article information

                Contributors
                tilman.schaeffer@uni-tuebingen.de
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                6 July 2017
                6 July 2017
                2017
                : 7
                : 4810
                Affiliations
                [1 ]ISNI 0000 0001 2190 1447, GRID grid.10392.39, Institute of Applied Physics, , University of Tübingen, ; Tübingen, Germany
                [2 ]ISNI 0000 0001 2190 1447, GRID grid.10392.39, Department of Physiology, , University of Tübingen, ; Tübingen, Germany
                [3 ]ISNI 0000 0001 2190 1447, GRID grid.10392.39, Department of Cardiology and Cardiovascular Diseases, , University of Tübingen, ; Tübingen, Germany
                Author information
                http://orcid.org/0000-0003-2962-1540
                Article
                4999
                10.1038/s41598-017-04999-6
                5500533
                28684746
                eb03e47c-54e8-4efc-a0bf-a6451f13b339
                © The Author(s) 2017

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 14 October 2016
                : 19 May 2017
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