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, Catalina, Masip, Martín E., Bragas, Andrea Verónica
Publicado:	2017
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:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00346748_v88_n1_p_VonBilderling http://hdl.handle.net/20.500.12110/paper_00346748_v88_n1_p_VonBilderling
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00346748_v88_n1_p_VonBilderling
record_format	dspace
spelling	paper:paper_00346748_v88_n1_p_VonBilderling2023-06-08T15:00:55Z Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus von Bilderling, Catalina Masip, Martín E. Bragas, Andrea Verónica 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. 2017 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00346748_v88_n1_p_VonBilderling 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, Catalina Masip, Martín E. Bragas, Andrea Verónica 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).
author	von Bilderling, Catalina Masip, Martín E. Bragas, Andrea Verónica
author_facet	von Bilderling, Catalina Masip, Martín E. Bragas, Andrea Verónica
author_sort	von Bilderling, Catalina
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
publishDate	2017
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00346748_v88_n1_p_VonBilderling http://hdl.handle.net/20.500.12110/paper_00346748_v88_n1_p_VonBilderling
work_keys_str_mv	AT vonbilderlingcatalina monitoringinrealtimefocaladhesionproteindynamicsinresponsetoadiscretemechanicalstimulus AT masipmartine monitoringinrealtimefocaladhesionproteindynamicsinresponsetoadiscretemechanicalstimulus AT bragasandreaveronica monitoringinrealtimefocaladhesionproteindynamicsinresponsetoadiscretemechanicalstimulus
_version_	1768544356936974336

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

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