Abstract:
The adhesion of cells to the extracellular matrix is a hierarchical, force-dependent, multistage process that evolves at several temporal scales. An understanding of this complex process requires a precise measurement of forces and its correlation with protein responses in living cells. We present a method to quantitatively assess live cell responses to a local and specific mechanical stimulus. Our approach combines atomic force microscopy with fluorescence imaging. Using this approach, we evaluated the recruitment of adhesion proteins such as vinculin, focal adhesion kinase, paxillin, and zyxin triggered by applying forces in the nN regime to live cells. We observed in real time the development of nascent adhesion sites, evident from the accumulation of early adhesion proteins at the position where the force was applied. We show that the method can be used to quantify the recruitment characteristic times for adhesion proteins in the formation of focal complexes. We also found a spatial remodeling of the mature focal adhesion protein zyxin as a function of the applied force. Our approach allows the study of a variety of complex biological processes involved in cellular mechanotransduction. © 2017 Author(s).
Registro:
Documento: |
Artículo
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Título: | Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus |
Autor: | Von Bilderling, C.; Caldarola, M.; Masip, M.E.; Bragas, A.V.; Pietrasanta, L.I. |
Filiación: | Centro de Microscopías Avanzadas, Departamento de Física, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina IFIBA-CONICET-UBA, Buenos Aires, Argentina Laboratorio de Electrónica Cuántica, Departamento de Física, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina Leiden Institute of Physics, Leiden University, Netherlands Max Planck Institute of Molecular Physiology, Dortmund, Germany
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Palabras clave: | Atomic force microscopy; Cells; Cytology; Proteins; Biological process; Extracellular matrices; Fluorescence imaging; Focal adhesion kinase; Mechanical stimulus; Mechanotransduction; Multistage process; Precise measurements; Adhesion; protein; cell function; chemistry; focal adhesion; mechanotransduction; Cell Physiological Phenomena; Focal Adhesions; Mechanotransduction, Cellular; Proteins |
Año: | 2017
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Volumen: | 88
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Número: | 1
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DOI: |
http://dx.doi.org/10.1063/1.4973664 |
Título revista: | Review of Scientific Instruments
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Título revista abreviado: | Rev. Sci. Instrum.
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ISSN: | 00346748
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CODEN: | RSINA
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CAS: | protein, 67254-75-5; Proteins
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Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00346748_v88_n1_p_VonBilderling |
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Citas:
---------- APA ----------
Von Bilderling, C., Caldarola, M., Masip, M.E., Bragas, A.V. & Pietrasanta, L.I.
(2017)
. Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus. Review of Scientific Instruments, 88(1).
http://dx.doi.org/10.1063/1.4973664---------- CHICAGO ----------
Von Bilderling, C., Caldarola, M., Masip, M.E., Bragas, A.V., Pietrasanta, L.I.
"Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus"
. Review of Scientific Instruments 88, no. 1
(2017).
http://dx.doi.org/10.1063/1.4973664---------- MLA ----------
Von Bilderling, C., Caldarola, M., Masip, M.E., Bragas, A.V., Pietrasanta, L.I.
"Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus"
. Review of Scientific Instruments, vol. 88, no. 1, 2017.
http://dx.doi.org/10.1063/1.4973664---------- VANCOUVER ----------
Von Bilderling, C., Caldarola, M., Masip, M.E., Bragas, A.V., Pietrasanta, L.I. Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus. Rev. Sci. Instrum. 2017;88(1).
http://dx.doi.org/10.1063/1.4973664