Absence of filamin A prevents cells from responding to stiffness gradients on gels coated with collagen but not fibronectin

Fitzroy J. Byfield, Qi Wen, Ilya Levental, Kerstin Nordstrom, Paulo E. Arratia, R. Tyler Miller, Paul A. Janmey

Research output: Contribution to journalArticle

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Abstract

Cell types from many tissues respond to changes in substrate stiffness by actively remodeling their cytoskeletons to alter spread area or adhesion strength, and in some cases changing their own stiffness to match that of their substrate. These cell responses to substrate stiffness are linked to substrate-induced changes in the state, localization, and amount of numerous proteins, but detailed evidence for the requirement of specific proteins in these distinct forms of mechanical response are scarce. Here we use microfluidics techniques to produce gels with a gradient of stiffness to show the essential function of filamin A in cell responses to mechanical stimuli and dissociate cell spreading and stiffening by contrasting responses of a pair of human melanoma-derived cell lines that differ in expression of this actin cross-linking protein. M2 melanoma cells null for filamin A do not alter their adherent area in response to increased substrate stiffness when they link to the substrate only through collagen receptors, but change adherent area normally when bound through fibronectin receptors. In contrast, filamin A-replete A7 cells change adherent area on both substrates and respond more strongly to collagen I-coated gels than to fibronectin-coated gels. Strikingly, A7 cells alter their stiffness, as measured by atomic force microscopy, to match the elastic modulus of the substrate immediately adjacent to them on the gradient. M2 cells, in contrast, maintain a constant stiffness on all substrates that is as low as that of A7 cells on the softest gels examined (1000 Pa). Comparison of cell spreading and cell stiffening on the same gradient substrates shows that cell spreading is uncoupled from stiffening. At saturating collagen and fibronectin concentrations, adhesion of M2 cells is reduced compared to that of A7 cells to an extent approximately equal to the difference in adherent area. Filamin A appears to be essential for cell stiffening on collagen, but not for cell spreading on fibronectin. These results have implications for different models of cell protrusion and adhesion and identify a key role for filamin A in altering cellular stiffness that cannot be compensated for by other actin cross-linkers in vivo.

Original languageEnglish (US)
Pages (from-to)5095-5102
Number of pages8
JournalBiophysical Journal
Volume96
Issue number12
DOIs
StatePublished - 2009

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Filamins
Fibronectins
Collagen
Gels
Cell Adhesion
Actins
Melanoma
Fibronectin Receptors
Collagen Receptors
Null Lymphocytes
Proteins
Microfluidics
Atomic Force Microscopy
Elastic Modulus

ASJC Scopus subject areas

  • Biophysics

Cite this

Absence of filamin A prevents cells from responding to stiffness gradients on gels coated with collagen but not fibronectin. / Byfield, Fitzroy J.; Wen, Qi; Levental, Ilya; Nordstrom, Kerstin; Arratia, Paulo E.; Miller, R. Tyler; Janmey, Paul A.

In: Biophysical Journal, Vol. 96, No. 12, 2009, p. 5095-5102.

Research output: Contribution to journalArticle

Byfield, Fitzroy J. ; Wen, Qi ; Levental, Ilya ; Nordstrom, Kerstin ; Arratia, Paulo E. ; Miller, R. Tyler ; Janmey, Paul A. / Absence of filamin A prevents cells from responding to stiffness gradients on gels coated with collagen but not fibronectin. In: Biophysical Journal. 2009 ; Vol. 96, No. 12. pp. 5095-5102.
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