Traction force microscopy in <i>Dictyostelium</i> reveals distinct roles for myosin II motor and actin-crosslinking activity in polarized cell movement

Lombardi, Maria L.; Knecht, David A.; Dembo, Micah; Lee, Juliet

doi:10.1242/jcs.002527

Cited by 94 publications

(97 citation statements)

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“…They claimed that myosin II motor and actin crosslinking activity were both required to develop and maintain asymmetrical patterns of traction stress, but they found that asymmetrical distributions sometimes even developed in myosin-II-null cells. These findings have led us to raise two questions: first, are there rearward traction stresses at the front of cells, as suggested by Uchida et al (Uchida et al, 2003), Tsujioka et al (Tsujioka et al, 2004), Ladam et al (Ladam et al, 2005) and del Alamo et al (del Alamo et al, 2007), or not, as suggested by Lombardi et al (Lombardi et al, 2007)? Second, if not, then what is the source of myosin-II-independent traction stress at the rear of myosin-II-null cells?…”

Section: Introductionmentioning

confidence: 95%

“…Tsujioka and colleagues (Tsujioka et al, 2004) also detected rearward traction stresses in migrating Dictyostelium cells. Recently, Lombardi et al (Lombardi et al, 2007) mapped traction stresses in migrating vegetative Dictyostelium cells. They found a lack of traction stress at the front but high stress at the rear of the cells, in contrast to Uchida et al (Uchida et al, 2003), It has been suggested that myosin II exerts traction forces at the posterior ends and retracting pseudopodia of migrating cells, but there is no direct evidence.…”

Section: Introductionmentioning

confidence: 99%

“…Lombardi et al (Lombardi et al, 2007) suggested that this traction stress asymmetry induced directed cell movement, and that the degree of this asymmetry decreased in cells null for the myosin II essential light chain (mlcE -) and was additionally decreased in myosin-II-null cells (mhcA -). They claimed that myosin II motor and actin crosslinking activity were both required to develop and maintain asymmetrical patterns of traction stress, but they found that asymmetrical distributions sometimes even developed in myosin-II-null cells.…”

Section: Introductionmentioning

confidence: 99%

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Actin-based propulsive forces and myosin-II-based contractile forces in migratingDictyosteliumcells

Iwadate

Yumura

2008

Journal of Cell Science

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It has been suggested that myosin II exerts traction forces at the posterior ends and retracting pseudopodia of migrating cells, but there is no direct evidence. Here, using a combination of total internal reflection fluorescence (TIRF) microscopy and force microscopy with a high spatial resolution of ∼400 nm, we simultaneously recorded GFP-myosin II dynamics and traction forces under migrating Dictyostelium cells. Accumulation of filamentous myosin II and a subsequent increase in traction forces were detected in pseudopodia just before retraction. In the case of motorless myosin II, traction forces did not increase after accumulation, suggesting that the source of the retraction force is the motor activity of accumulated myosin II. Simultaneous recording of F-actin and traction forces revealed that traction forces were exerted under spot-like regions where F-actin accumulated. Cells migrated in a direction counter to the sum of the force vectors exerted at each spot, suggesting that the stress spots act as scaffolds to transmit the propulsive forces at the leading edge generated by actin polymerization.

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Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Actin-based propulsive forces and myosin-II-based contractile forces in migratingDictyosteliumcells

Iwadate

Yumura

2008

Journal of Cell Science

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“…The maximum force of large fibroblasts is several tens of kilopascals (Dembo and Wang, 1999;Munevar et al, 2001aMunevar et al, , 2001b, although that of small cell types, such as Dictyostelium cells (Iwadate and Yumura, 2008a;Lombardi et al, 2007;Tanimoto and Sano, 2014), is several kilopascals or smaller. The magnitude of the traction forces may be related to cell size.…”

Section: Introductionmentioning

confidence: 99%

Shape and Area of Keratocytes Are Related to the Distribution and Magnitude of Their Traction Forces

Sonoda

Okimura

Iwadate

2016

Cell Struct. Funct.

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ABSTRACT. Fish epidermal keratocytes maintain an overall fan shape during their crawling migration. The shape-determination mechanism has been described theoretically and experimentally on the basis of graded radial extension of the leading edge, but the relationship between shape and traction forces has not been clarified. Migrating keratocytes can be divided into fragments by treatment with the protein kinase inhibitor staurosporine. Fragments containing a nucleus and cytoplasm behave as mini-keratocytes and maintain the same fan shape as the original cells. We measured the shape of the leading edge, together with the areas of the ventral region and traction forces, of keratocytes and mini-keratocytes. The shapes of keratocytes and mini-keratocytes were similar. Mini-keratocytes exerted traction forces at the rear left and right ends, just like keratocytes. The magnitude of the traction forces was proportional to the area of the keratocytes and mini-keratocytes. The myosin II ATPase inhibitor blebbistatin decreased the forces at the rear left and right ends of the keratocytes and expanded their shape laterally. These results suggest that keratocyte shape depends on the distribution of the traction forces, and that the magnitude of the traction forces depends on the area of the cells.

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“…Use of a recently developed digital volume correlation (DVC) method (16) allows 3-D displacements and traction fields to be determined directly from volumetric confocal image stacks, and obviates the need for complex inverse formulations (10). The method has a temporal resolution that permits confocal imaging over time scales relevant for the migration of anchorage dependent cells, such as endothelial cells and fibroblasts (17).…”

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confidence: 99%

Quantifying cellular traction forces in three dimensions

Maskarinec

Franck

Tirrell

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

264

224

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Cells engage in mechanical force exchange with their extracellular environment through tension generated by the cytoskeleton. A method combining laser scanning confocal microscopy (LSCM) and digital volume correlation (DVC) enables tracking and quantification of cell-mediated deformation of the extracellular matrix in all three spatial dimensions. Time-lapse confocal imaging of migrating 3T3 fibroblasts on fibronectin (FN)-modified polyacrylamide gels of varying thickness reveals significant in-plane (x, y) and normal (z) displacements, and illustrates the extent to which cells, even in nominally two-dimensional (2-D) environments, explore their surroundings in all three dimensions. The magnitudes of the measured displacements are independent of the elastic moduli of the gels. Analysis of the normal displacement profiles suggests that normal forces play important roles even in 2-D cell migration.digital volume correlation ͉ laser scanning confocal microscopy ͉ three-dimensional T he measurement of cellular traction forces has been of increasing interest since the discovery that the mechanical properties of the cellular microenvironment can direct many important cellular processes including spreading, migration, and differentiation (1-4). It is now widely accepted that mechanical properties must be considered along with chemical signals if we are to understand how cells integrate environmental cues to modulate their behavior (5-8). The correlation between cell-induced deformations in materials and biochemical signaling and regulation, particularly focal adhesion formation and clustering, has been investigated through the use of a variety of techniques including surface wrinkling, displacement-tracking using traction force microscopy (TFM), and bending of pillar arrays (9-15). These methods have yielded substantial insight into cellular behavior, but are inherently restricted to two-dimensional (2-D) analysis and interpretation of cell-matrix interactions. Furthermore, these approaches calculate stresses by comparing images before and after cell detachment (10), thus providing only snapshots of cell behavior rather than dynamic analyses of the processes by which cells explore their microenvironments.In this report, we demonstrate the capability to dynamically track and quantify cellular traction forces in three dimensions (3-D). Mechanical interactions between 3T3 fibroblasts and FN-modified polyacrylamide gels are quantified dynamically by computing the displacement and traction fields generated by motile cells. Use of a recently developed digital volume correlation (DVC) method (16) allows 3-D displacements and traction fields to be determined directly from volumetric confocal image stacks, and obviates the need for complex inverse formulations (10). The method has a temporal resolution that permits confocal imaging over time scales relevant for the migration of anchorage dependent cells, such as endothelial cells and fibroblasts (17).The 3-D character of this approach relies on the use of laser scanning confoca...

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Traction force microscopy in Dictyostelium reveals distinct roles for myosin II motor and actin-crosslinking activity in polarized cell movement

Cited by 94 publications

References 46 publications

Actin-based propulsive forces and myosin-II-based contractile forces in migratingDictyosteliumcells

Actin-based propulsive forces and myosin-II-based contractile forces in migratingDictyosteliumcells

Shape and Area of Keratocytes Are Related to the Distribution and Magnitude of Their Traction Forces

Quantifying cellular traction forces in three dimensions

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