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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
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
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
Volumen:88
Número:1
DOI: http://dx.doi.org/10.1063/1.4973664
Título revista:Review of Scientific Instruments
Título revista abreviado:Rev. Sci. Instrum.
ISSN:00346748
CODEN:RSINA
CAS:protein, 67254-75-5; Proteins
Registro:http://digital.bl.fcen.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