Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus

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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...

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Autores principales: Von Bilderling, C., Caldarola, M., Masip, M.E., Bragas, A.V., Pietrasanta, L.I.
Formato: JOUR
Materias:
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
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00346748_v88_n1_p_VonBilderling
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Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id todo:paper_00346748_v88_n1_p_VonBilderling
record_format dspace
spelling todo:paper_00346748_v88_n1_p_VonBilderling2023-10-03T14:46:11Z Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus Von Bilderling, C. Caldarola, M. Masip, M.E. Bragas, A.V. Pietrasanta, L.I. 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 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). Fil:Von Bilderling, C. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Masip, M.E. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Bragas, A.V. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00346748_v88_n1_p_VonBilderling
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic 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
spellingShingle 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
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
topic_facet 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
description 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).
format JOUR
author Von Bilderling, C.
Caldarola, M.
Masip, M.E.
Bragas, A.V.
Pietrasanta, L.I.
author_facet Von Bilderling, C.
Caldarola, M.
Masip, M.E.
Bragas, A.V.
Pietrasanta, L.I.
author_sort Von Bilderling, C.
title Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
title_short Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
title_full Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
title_fullStr Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
title_full_unstemmed Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
title_sort monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
url http://hdl.handle.net/20.500.12110/paper_00346748_v88_n1_p_VonBilderling
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AT masipme monitoringinrealtimefocaladhesionproteindynamicsinresponsetoadiscretemechanicalstimulus
AT bragasav monitoringinrealtimefocaladhesionproteindynamicsinresponsetoadiscretemechanicalstimulus
AT pietrasantali monitoringinrealtimefocaladhesionproteindynamicsinresponsetoadiscretemechanicalstimulus
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