Traction forces of fibroblasts are regulated by the Rho-dependent kinase but not by the myosin light chain kinase

Beningo, Karen A.; Hamao, Kozue; Dembo, Micah; Wang, Yu Li; Hosoya, Hiroshi

doi:10.1016/j.abb.2006.09.025

Cited by 115 publications

(107 citation statements)

References 51 publications

(67 reference statements)

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“…Because the bulk of traction forces have been shown to be generated by myosin-II-mediated contractility (Beningo et al, 2006), we expected blebbistatin, a potent inhibitor of myosin II ATPase, to inhibit not only traction forces, but also any increase in traction force upon microtubule depolymerization. Blebbistatin treatment caused a sixfold decrease in average traction stress in patterned cells.…”

Section: Resultsmentioning

confidence: 99%

“…Previous studies have indicated a strong dependence of traction forces on myosin II (Beningo et al, 2006;Gardel et al, 2008). Patterning cells allows more precise quantification and has indicated an incomplete inhibition of traction stress upon blebbistatin treatment.…”

Section: Discussionmentioning

confidence: 99%

“…The production of traction forces -forces that are transmitted from the actin cytoskeleton through focal adhesions to the substrate (Harris et al, 1980) -is necessary for the cell to perform these vital tasks. The actin cytoskeleton, in conjunction with the motor protein myosin II, is responsible for the production of traction forces (Pelham and Wang, 1999;Beningo et al, 2006). Probably due to the association of many signaling factors with cytoskeletal elements, proper regulation of traction forces is essential for many mechanosensing phenomena, including the responses of cell migration, growth and differentiation to substrate compliance (Engler et al, 2006;Lo et al, 2004;Ulrich et al, 2009) and to cell shape (McBeath et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

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Microtubule depolymerization induces traction force increase through two distinct pathways

Rape

Guo

Wang

2011

Journal of Cell Science

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SummaryTraction forces increase after microtubule depolymerization; however, the signaling mechanisms underlying this, in particular the dependence upon myosin II, remain unclear. We investigated the mechanism of traction force increase after nocodazole-induced microtubule depolymerization by applying traction force microscopy to cells cultured on micropatterned polyacrylamide hydrogels to obtain samples of homogeneous shape and size. Control cells and cells treated with a focal adhesion kinase (FAK) inhibitor showed similar increases in traction forces, indicating that the response is independent of FAK. Surprisingly, pharmacological inhibition of myosin II did not prevent the increase of residual traction forces upon nocodazole treatment. This increase was abolished upon pharmacological inhibition of FAK. These results suggest two distinct pathways for the regulation of traction forces. First, microtubule depolymerization activates a myosin-II-dependent mechanism through a FAK-independent pathway. Second, microtubule depolymerization also enhances traction forces through a myosin-II-independent, FAK-regulated pathway. Traction forces are therefore regulated by a complex network of complementary signals and force-generating mechanisms.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Microtubule depolymerization induces traction force increase through two distinct pathways

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Guo

Wang

2011

Journal of Cell Science

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“…Therefore, mechanical forces, either generated internally by myosin II or applied externally through focal adhesions, may play a role in inducing the fluxes. Y-27632, an inhibitor of the Rho-dependent kinase, and blebbistatin, an inhibitor of myosin II ATPase, both caused strong inhibition of traction forces (Beningo et al, 2006), and focal adhesion fluxes, while reducing zyxin tails to small dot-like structures (Figure 6, A-C, and Supplemental Movie 6). Conversely, application of pulling forces to cells plated on flexible polyacrylamide substrates stimulated the appearance of focal adhesion fluxes (Figure 6, D-G).…”

Section: Dependence Of Focal Adhesion Fluxes On Cellular Contractilitmentioning

confidence: 99%

Retrograde Fluxes of Focal Adhesion Proteins in Response to Cell Migration and Mechanical Signals

Guo

Wang

2007

MBoC

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Recent studies suggest that mechanical signals mediated by the extracellular matrix play an essential role in various physiological and pathological processes; yet, how cells respond to mechanical stimuli remains elusive. Using live cell fluorescence imaging, we found that actin filaments, in association with a number of focal adhesion proteins, including zyxin and vasodilator-stimulated phosphoprotein, undergo retrograde fluxes at focal adhesions in the lamella region. This flux is inversely related to cell migration, such that it is amplified in fibroblasts immobilized on micropatterned islands. In addition, the flux is regulated by mechanical signals, including stretching forces applied to flexible substrates and substrate stiffness. Conditions favoring the flux share the common feature of causing large retrograde displacements of the interior actin cytoskeleton relative to the substrate anchorage site, which may function as a switch translating mechanical input into chemical signals, such as tyrosine phosphorylation. In turn, the stimulation of actin flux at focal adhesions may function as part of a feedback mechanism, regulating structural assembly and force production in relation to cell migration and mechanical load. The retrograde transport of associated focal adhesion proteins may play additional roles in delivering signals from focal adhesions to the interior of the cell. INTRODUCTIONRecent studies suggest that mechanosensing is involved in several physiological processes, including embryogenesis (Newman and Comper, 1990;Beloussov et al., 1994) and wound healing (Hinz et al., 2001;Tomasek et al., 2002), and in pathological processes such as fibrosis and carcinogenesis (Paszek et al., 2005). Adherent cells seem capable of both responding to applied mechanical forces (Tzima et al., 2005) and applying contractile forces to probe mechanical properties of the environment (Discher et al., 2005). The downstream responses include changes in migration (Pelham and Wang, 1997;Sheetz et al., 1998;Lo et al., 2000), cell-cell interactions (Guo et al., 2006), proliferation (Wang et al., 2000Nelson et al., 2005), differentiation (Engler et al., 2004(Engler et al., , 2006, and apoptosis . For cultured adherent cells, focal adhesions are thought to mediate mechanosensing through integrin-mediated anchorage to the extracellular matrix (Larsen et al., 2006). The molecular repertoire of focal adhesions includes Ͼ100 adaptor and signaling proteins (Bershadsky et al., 2006), in addition to associated actomyosin bundles that provide mechanical forces for probing the environment (Choquet et al., 1997), inside-out signaling (Chrzanowska-Wodnicka and Burridge, 1996;Schwartz and Ginsberg, 2002), and cell migration (Ridley et al., 2003). However, few details are available concerning how these components work in concert to generate the responses to mechanical signals.Several aspects have emerged recently as the key features of integrin-mediated mechanosensing. First is the increase in cortical organization and contractility in resp...

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“…Cells respond to outside-in signals by exerting actomyosin-based contractile forces on the matrix (inside-out forces) that increase cell stiffness and scale with ECM stiffness [2]. Rho/ROCK signalling is rapidly activated at ECM adhesions in response to matrix stiffness to augment actomyosin activity, via actin polymerisation and myosin light chain phosphorylation, and increase cell stiffness [3,4].…”

Section: Introductionmentioning

confidence: 99%

The Use of Polyacrylamide Hydrogels to Study the Effects of Matrix Stiffness on Nuclear Envelope Properties

Minaisah

Cox

Warren

2016

Methods in Molecular Biology

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Traction forces of fibroblasts are regulated by the Rho-dependent kinase but not by the myosin light chain kinase

Cited by 115 publications

References 51 publications

Microtubule depolymerization induces traction force increase through two distinct pathways

Microtubule depolymerization induces traction force increase through two distinct pathways

Retrograde Fluxes of Focal Adhesion Proteins in Response to Cell Migration and Mechanical Signals

The Use of Polyacrylamide Hydrogels to Study the Effects of Matrix Stiffness on Nuclear Envelope Properties

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