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

Autores
Von Bilderling, Catalina; Caldarola, Martín; Masip, Martín E.; Bragas, Andrea Veronica; Pietrasanta, Lia
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The adhesion of cells to the extracellular matrix is a hierarchical, force-dependent, multistage processthat evolves at several temporal scales. An understanding of this complex process requires a precisemeasurement of forces and its correlation with protein responses in living cells. We present a methodto quantitatively assess live cell responses to a local and specific mechanical stimulus. Our approachcombines atomic force microscopy with fluorescence imaging. Using this approach, we evaluatedthe recruitment of adhesion proteins such as vinculin, focal adhesion kinase, paxillin, and zyxintriggered by applying forces in the nN regime to live cells. We observed in real time the developmentof nascent adhesion sites, evident from the accumulation of early adhesion proteins at the positionwhere the force was applied. We show that the method can be used to quantify the recruitmentcharacteristic times for adhesion proteins in the formation of focal complexes. We also found aspatial 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 cellularmechanotransduction
Fil: Von Bilderling, Catalina. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Caldarola, Martín. Leiden University; Países Bajos. Universidad de Buenos Aires; Argentina
Fil: Masip, Martín E.. Universidad de Buenos Aires; Argentina. Institut Max Planck fur Molekulare Physiologie; Alemania
Fil: Bragas, Andrea Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; Argentina
Fil: Pietrasanta, Lia. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Materia
Focal Adhesions
Mechanotransduction
Optical Microscope
Afm
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/48990
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/48990
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulusVon Bilderling, CatalinaCaldarola, MartínMasip, Martín E.Bragas, Andrea VeronicaPietrasanta, LiaFocal AdhesionsMechanotransductionOptical MicroscopeAfmhttps://purl.org/becyt/ford/1.3https://purl.org/becyt/ford/1The adhesion of cells to the extracellular matrix is a hierarchical, force-dependent, multistage processthat evolves at several temporal scales. An understanding of this complex process requires a precisemeasurement of forces and its correlation with protein responses in living cells. We present a methodto quantitatively assess live cell responses to a local and specific mechanical stimulus. Our approachcombines atomic force microscopy with fluorescence imaging. Using this approach, we evaluatedthe recruitment of adhesion proteins such as vinculin, focal adhesion kinase, paxillin, and zyxintriggered by applying forces in the nN regime to live cells. We observed in real time the developmentof nascent adhesion sites, evident from the accumulation of early adhesion proteins at the positionwhere the force was applied. We show that the method can be used to quantify the recruitmentcharacteristic times for adhesion proteins in the formation of focal complexes. We also found aspatial 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 cellularmechanotransductionFil: Von Bilderling, Catalina. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Caldarola, Martín. Leiden University; Países Bajos. Universidad de Buenos Aires; ArgentinaFil: Masip, Martín E.. Universidad de Buenos Aires; Argentina. Institut Max Planck fur Molekulare Physiologie; AlemaniaFil: Bragas, Andrea Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; ArgentinaFil: Pietrasanta, Lia. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaAmerican Institute of Physics2017-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/48990Von Bilderling, Catalina; Caldarola, Martín; Masip, Martín E.; Bragas, Andrea Veronica; Pietrasanta, Lia; Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus; American Institute of Physics; Review of Scientific Instruments; 88; 1; 1-2017; 1-90034-6748CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1063/1.4973664info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:50:33Zoai:ri.conicet.gov.ar:11336/48990instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:50:33.877CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
title Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
spellingShingle Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus
Von Bilderling, Catalina
Focal Adhesions
Mechanotransduction
Optical Microscope
Afm
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
dc.creator.none.fl_str_mv Von Bilderling, Catalina
Caldarola, Martín
Masip, Martín E.
Bragas, Andrea Veronica
Pietrasanta, Lia
author Von Bilderling, Catalina
author_facet Von Bilderling, Catalina
Caldarola, Martín
Masip, Martín E.
Bragas, Andrea Veronica
Pietrasanta, Lia
author_role author
author2 Caldarola, Martín
Masip, Martín E.
Bragas, Andrea Veronica
Pietrasanta, Lia
author2_role author
author
author
author
dc.subject.none.fl_str_mv Focal Adhesions
Mechanotransduction
Optical Microscope
Afm
topic Focal Adhesions
Mechanotransduction
Optical Microscope
Afm
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.3
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The adhesion of cells to the extracellular matrix is a hierarchical, force-dependent, multistage processthat evolves at several temporal scales. An understanding of this complex process requires a precisemeasurement of forces and its correlation with protein responses in living cells. We present a methodto quantitatively assess live cell responses to a local and specific mechanical stimulus. Our approachcombines atomic force microscopy with fluorescence imaging. Using this approach, we evaluatedthe recruitment of adhesion proteins such as vinculin, focal adhesion kinase, paxillin, and zyxintriggered by applying forces in the nN regime to live cells. We observed in real time the developmentof nascent adhesion sites, evident from the accumulation of early adhesion proteins at the positionwhere the force was applied. We show that the method can be used to quantify the recruitmentcharacteristic times for adhesion proteins in the formation of focal complexes. We also found aspatial 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 cellularmechanotransduction
Fil: Von Bilderling, Catalina. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Caldarola, Martín. Leiden University; Países Bajos. Universidad de Buenos Aires; Argentina
Fil: Masip, Martín E.. Universidad de Buenos Aires; Argentina. Institut Max Planck fur Molekulare Physiologie; Alemania
Fil: Bragas, Andrea Veronica. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de Buenos Aires; Argentina
Fil: Pietrasanta, Lia. Universidad de Buenos Aires; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
description The adhesion of cells to the extracellular matrix is a hierarchical, force-dependent, multistage processthat evolves at several temporal scales. An understanding of this complex process requires a precisemeasurement of forces and its correlation with protein responses in living cells. We present a methodto quantitatively assess live cell responses to a local and specific mechanical stimulus. Our approachcombines atomic force microscopy with fluorescence imaging. Using this approach, we evaluatedthe recruitment of adhesion proteins such as vinculin, focal adhesion kinase, paxillin, and zyxintriggered by applying forces in the nN regime to live cells. We observed in real time the developmentof nascent adhesion sites, evident from the accumulation of early adhesion proteins at the positionwhere the force was applied. We show that the method can be used to quantify the recruitmentcharacteristic times for adhesion proteins in the formation of focal complexes. We also found aspatial 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 cellularmechanotransduction
publishDate 2017
dc.date.none.fl_str_mv 2017-01
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/48990
Von Bilderling, Catalina; Caldarola, Martín; Masip, Martín E.; Bragas, Andrea Veronica; Pietrasanta, Lia; Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus; American Institute of Physics; Review of Scientific Instruments; 88; 1; 1-2017; 1-9
0034-6748
CONICET Digital
CONICET
url http://hdl.handle.net/11336/48990
identifier_str_mv Von Bilderling, Catalina; Caldarola, Martín; Masip, Martín E.; Bragas, Andrea Veronica; Pietrasanta, Lia; Monitoring in real-time focal adhesion protein dynamics in response to a discrete mechanical stimulus; American Institute of Physics; Review of Scientific Instruments; 88; 1; 1-2017; 1-9
0034-6748
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1063/1.4973664
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Institute of Physics
publisher.none.fl_str_mv American Institute of Physics
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844613558067789824
score 13.070432

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