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Quantitative Phase Imaging of Spreading Fibroblasts Identifies the Role of Focal Adhesion Kinase in the Stabilization of the Cell Rear
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SYSNO ASEP 0533929 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Quantitative Phase Imaging of Spreading Fibroblasts Identifies the Role of Focal Adhesion Kinase in the Stabilization of the Cell Rear Author(s) Ramaniuk, Olga (MBU-M) ORCID
Klímová, Zuzana (MBU-M) RID
Groušl, Tomáš (MBU-M) RID, ORCID
Vomastek, Tomáš (MBU-M) RID, ORCIDArticle number 1089 Source Title Biomolecules. - : MDPI
Roč. 10, č. 8 (2020)Number of pages 23 s. Language eng - English Country CH - Switzerland Keywords coherence-controlled holographic microscopy ; quantitative phase imaging ; cell dry mass Subject RIV EE - Microbiology, Virology OECD category Microbiology R&D Projects GJ19-07603Y GA ČR - Czech Science Foundation (CSF) GA18-11908S GA ČR - Czech Science Foundation (CSF) LM2018129 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support MBU-M - RVO:61388971 UT WOS 000578864200001 EID SCOPUS 85088288037 DOI 10.3390/biom10081089 Annotation Cells attaching to the extracellular matrix spontaneously acquire front-rear polarity. This self-organization process comprises spatial activation of polarity signaling networks and the establishment of a protruding cell front and a non-protruding cell rear. Cell polarization also involves the reorganization of cell mass, notably the nucleus that is positioned at the cell rear. It remains unclear, however, how these processes are regulated. Here, using coherence-controlled holographic microscopy (CCHM) for non-invasive live-cell quantitative phase imaging (QPI), we examined the role of the focal adhesion kinase (FAK) and its interacting partner Rack1 in dry mass distribution in spreading Rat2 fibroblasts. We found that FAK-depleted cells adopt an elongated, bipolar phenotype with a high central body mass that gradually decreases toward the ends of the elongated processes. Further characterization of spreading cells showed that FAK-depleted cells are incapable of forming a stable rear, rather, they form two distally positioned protruding regions. Continuous protrusions at opposite sides results in an elongated cell shape. In contrast, Rack1-depleted cells are round and large with the cell mass sharply dropping from the nuclear area towards the basal side. We propose that FAK and Rack1 act differently yet coordinately to establish front-rear polarity in spreading cells. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2021 Electronic address https://www.mdpi.com/2218-273X/10/8/1089
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