scholarly journals Discussion of “Cytoskeletal Mechanics Regulating Amoeboid Cell Locomotion” (Álvarez-González, B., Bastounis, E., Meili, R., del Alamo, J. C., Firtel, R. A., and Lasheras, J. C., 2014, ASME Appl. Mech. Rev., 66(5), p. 050804)

2014 ◽  
Vol 66 (5) ◽  
Author(s):  
Xavier Trepat
Keyword(s):  
Cell Motility ◽  
Traction Force ◽  
Traction Forces ◽  
Force Microscopy ◽  
Amoeboid Cell ◽  
Cell Locomotion ◽  
Cell Cycle Synchronization ◽  
Open Questions ◽  
Universal Feature ◽  
The Relationship

A virtually universal feature of adherent cells is their ability to exert traction forces. To measure these forces, several methods have been developed over the past 15 years. In this issue of Applied Mechanics Reviews, Álvarez-González and co-workers review their own traction force microscopy approach and its application to the study of amoeboid cell locomotion. They show that the cycle of cell motility is exquisitely synchronized by a cycle of traction forces. In addition, they show how traction forces and cell cycle synchronization are affected by myosin and SCAR/WAVE mutants. Here, I discuss some open questions that derive from the work of the authors and other laboratories as regards the relationship between cell motility and traction forces.

scholarly journals Both contractile axial and lateral traction force dynamics drive amoeboid cell motility

2014 ◽  
Vol 204 (6) ◽  
pp. 1045-1061 ◽  
Author(s):  
Effie Bastounis ◽  
Ruedi Meili ◽  
Begoña Álvarez-González ◽  
Joshua Francois ◽  
Juan C. del Álamo ◽  
...  
Keyword(s):  
Cell Motility ◽  
Traction Force ◽  
Force Microscopy ◽  
Force Dynamics ◽  
Amoeboid Cell ◽  
Axial Forces ◽  
Extrinsic Cues ◽  
Mutant Strains ◽  
And Migration ◽  
Anterior Posterior

Chemotaxing Dictyostelium discoideum cells adapt their morphology and migration speed in response to intrinsic and extrinsic cues. Using Fourier traction force microscopy, we measured the spatiotemporal evolution of shape and traction stresses and constructed traction tension kymographs to analyze cell motility as a function of the dynamics of the cell’s mechanically active traction adhesions. We show that wild-type cells migrate in a step-wise fashion, mainly forming stationary traction adhesions along their anterior–posterior axes and exerting strong contractile axial forces. We demonstrate that lateral forces are also important for motility, especially for migration on highly adhesive substrates. Analysis of two mutant strains lacking distinct actin cross-linkers (mhcA− and abp120− cells) on normal and highly adhesive substrates supports a key role for lateral contractions in amoeboid cell motility, whereas the differences in their traction adhesion dynamics suggest that these two strains use distinct mechanisms to achieve migration. Finally, we provide evidence that the above patterns of migration may be conserved in mammalian amoeboid cells.

scholarly journals A model for estimating traction force magnitude reveals differential regulation of actomyosin activity and matrix adhesion number in response to smooth muscle cell spreading

2019 ◽  
Author(s):  
Sultan Ahmed ◽  
Panashe Mabeza ◽  
Derek T Warren
Keyword(s):  
Smooth Muscle ◽  
Linear Regression Analysis ◽  
Traction Force ◽  
Strongly Correlated ◽  
Matrix Rigidity ◽  
Aortic Compliance ◽  
Force Microscopy ◽  
Force Magnitude ◽  
The Relationship ◽  
Novel Model

AbstractDecreased aortic compliance is associated with ageing and vascular disease, including atherosclerosis and hypertension. Ultimately, changes in aortic compliance are driven by altered ECM composition however, recent findings have identified a cellular component to decreased aortic compliance observed in ageing and hypertension. Vascular smooth muscle cells (VSMCs) line the blood vessel wall and VSMC contraction regulates vascular tone and contributes to aortic compliance. Mechanical cues derived from the ECM influence VSMC function, yet whether ECM rigidity influences VSMC force generation remains unclear. In this study, we describe the relationship between VSMC spreading, traction force magnitude and matrix rigidity. Importantly, we show that spreading predicts integrated traction force (integrated-TF) magnitude independently of matrix rigidity. Using linear regression analysis, we have generated a model for calculating integrated-TF from VSMC area. This model closely predicts the integrated traction force measured by live VSMC traction force microscopy. Vinculin staining analysis revealed that spreading strongly correlated with adhesion number per VSMC, suggesting that increased VSMC integrated-TF was due to enhanced matrix anchor points. Further analysis revealed that calculated integrated-TF per adhesion was reduced by matrix rigidity, however, adhesion number/μm2increased, resulting in the average integrated-TF/μm2remaining unaltered. As a result, the integrated-TF/VSMC spreading relationship is independent of matrix rigidity. Therefore, our study has identified and validated a novel model to predict and understand the mechanisms influencing VSMC traction force magnitude.

scholarly journals Black Dots: Microcontact-Printed, Reference-Free Traction Force Microscopy

2021 ◽  
Author(s):  
Kevin M Beussman ◽  
Molly Y Mollica ◽  
Andrea Leonard ◽  
Jeffrey Miles ◽  
John Hocter ◽  
...  
Keyword(s):  
Flexible Substrate ◽  
Traction Force ◽  
Human Platelets ◽  
High Yield ◽  
Traction Forces ◽  
Cooperative Effects ◽  
Force Microscopy ◽  
Interaction Terms ◽  
Spread Area ◽  
Insight Into

Measuring the traction forces produced by cells provides insight into their behavior and physiological function. Here, we developed a technique (dubbed 'black dots') that microcontact prints a fluorescent micropattern onto a flexible substrate to measure cellular traction forces without constraining cell shape or needing to detach the cells. To demonstrate our technique, we assessed human platelets, which can generate a large range of forces within a population. We find platelets that exert more force have more spread area, are more circular, and have more uniformly distributed F-actin filaments. As a result of the high yield of data obtainable by this technique, we were able to evaluate multivariate mixed effects models with interaction terms and conduct a clustering analysis to identify clusters within our data. These statistical techniques demonstrated a complex relationship between spread area, circularity, F-actin dispersion, and platelet force, including cooperative effects that significantly associate with platelet traction forces.

scholarly journals A Novel Method for Quantifying Traction Forces from Hexagonal Micropatterned Features on Deformable Poly-Dimethyl Siloxane Sheets

2018 ◽  
Author(s):  
Brian P. Griffin ◽  
Christopher J. Largaespada ◽  
Nicole A. Rinaldi ◽  
Christopher A. Lemmon
Keyword(s):  
Computational Algorithm ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Polydimethyl Siloxane ◽  
Traction Forces ◽  
Homogeneous Surface ◽  
Force Microscopy ◽  
Dimethyl Siloxane ◽  
Novel Method ◽  
Cellular Forces

AbstractMany methods exist for quantifying cellular traction forces, including traction force microscopy and microfabricated post arrays. However, these methodologies have limitations, including a requirement to remove cells to determine undeflected particle locations and the inability to quantify forces of cells with low cytoskeletal stiffness, respectively. Here we present a novel method of traction force quantification that eliminates both of these limitations. Through the use of a hexagonal pattern of microcontact-printed protein spots, a novel computational algorithm, and thin surfaces of polydimethyl siloxane (PDMS) blends, we demonstrate a system that quantifies cellular forces on a homogeneous surface that is stable, easily manufactured, and can quantify forces without need for cellular removal.

scholarly journals Three-Dimensional Quantification of Cellular Traction Forces and Mechanosensing of Thin Substrata by Fourier Traction Force Microscopy

PLoS ONE ◽  
2013 ◽  
Vol 8 (9) ◽  
pp. e69850 ◽  
Author(s):  
Juan C. del Álamo ◽  
Ruedi Meili ◽  
Begoña Álvarez-González ◽  
Baldomero Alonso-Latorre ◽  
Effie Bastounis ◽  
...  
Keyword(s):  
Three Dimensional ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Traction Forces ◽  
Force Microscopy

scholarly journals Quantifying force transmission through fibroblasts: changes of traction forces under external shearing

2021 ◽  
Author(s):  
Steven Huth ◽  
Johannes W. Blumberg ◽  
Dimitri Probst ◽  
Jan Lammerding ◽  
Ulrich S. Schwarz ◽  
...  
Keyword(s):  
Cell Surface ◽  
Mammalian Cells ◽  
Shear Force ◽  
Apical Cell ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Force Transmission ◽  
Traction Forces ◽  
Force Microscopy ◽  
Adhesion Sites

AbstractMammalian cells have evolved complex mechanical connections to their microenvironment, including focal adhesion clusters that physically connect the cytoskeleton and the extracellular matrix. This mechanical link is also part of the cellular machinery to transduce, sense and respond to external forces. Although methods to measure cell attachment and cellular traction forces are well established, these are not capable of quantifying force transmission through the cell body to adhesion sites. We here present a novel approach to quantify intracellular force transmission by combining microneedle shearing at the apical cell surface with traction force microscopy at the basal cell surface. The change of traction forces exerted by fibroblasts to underlying polyacrylamide substrates as a response to a known shear force exerted with a calibrated microneedle reveals that cells redistribute forces dynamically under external shearing and during sequential rupture of their adhesion sites. Our quantitative results demonstrate a transition from dipolar to monopolar traction patterns, an inhomogeneous distribution of the external shear force to the adhesion sites as well as dynamical changes in force loading prior to and after the rupture of single adhesion sites. Our strategy of combining traction force microscopy with external force application opens new perspectives for future studies of force transmission and mechanotransduction in cells.

scholarly journals The biochemical composition of the actomyosin network sets the magnitude of cellular traction forces

2021 ◽  
Vol 32 (18) ◽  
pp. 1737-1748
Author(s):  
Somanna Kollimada ◽  
Fabrice Senger ◽  
Timothée Vignaud ◽  
Manuel Théry ◽  
Laurent Blanchoin ◽  
...  
Keyword(s):  
Focal Adhesion ◽  
Biochemical Composition ◽  
Force Production ◽  
Traction Force ◽  
Traction Force Microscopy ◽  
Traction Forces ◽  
Force Microscopy ◽  
Cell Force

The endogenous content of proteins associated with force production and the resultant traction forces were quantified in the same cells using a new traction force-microscopy assay. Focal adhesion size correlated with force in stationary cells. Relative numbers of motors and cross-linkers per actin required an optimum to maximize cell force production.

scholarly journals Crosstalk between myosin II and formin functions in the regulation of force generation and actomyosin dynamics in stress fibers

2021 ◽  
Author(s):  
Yukako Nishimura ◽  
Shidong Shi ◽  
Qingsen Li ◽  
Alexander D. Bershadsky ◽  
Virgile Viasnoff
Keyword(s):  
Contractile Activity ◽  
Myosin Ii ◽  
Focal Adhesions ◽  
Traction Force ◽  
Stress Fiber ◽  
Force Generation ◽  
Stress Fibers ◽  
Force Microscopy ◽  
Fiber Dynamics ◽  
The Relationship

REF52 fibroblasts have a well-developed contractile machinery, the most prominent elements of which are actomyosin stress fibers with highly ordered organization of actin and myosin IIA filaments. The relationship between contractile activity and turnover dynamics of stress fibers is not sufficiently understood. Here, we simultaneously measured the forces exerted by stress fibers (using traction force microscopy or micropillar array sensors) and the dynamics of actin and myosin (using photoconversion-based monitoring of actin incorporation and high-resolution fluorescence microscopy of myosin II light chain). Our data revealed new features of the crosstalk between myosin II-driven contractility and stress fiber dynamics. During normal stress fiber turnover, actin incorporated all along the stress fibers and not only at focal adhesions. Incorporation of actin into stress fibers/focal adhesions, as well as actin and myosin II filaments flow along stress fibers, strongly depends on myosin II activity. Myosin II-dependent generation of traction forces does not depend on incorporation of actin into stress fibers per se, but still requires formin activity. This previously overlooked function of formins in maintenance of the actin cytoskeleton connectivity could be the main mechanism of formin involvement in traction force generation.

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