Spatio-temporal analysis of eukaryotic cell motility by improved force cytometry

Álamo, Juan C. del; Meili, Ruedi; Alonso-Latorre, Baldomero; Rodríguez-Rodríguez, Javier; Aliseda, Alberto; Firtel, Richard A.; Lasheras, Juan C.

doi:10.1073/pnas.0705815104

Cited by 195 publications

(335 citation statements)

References 35 publications

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“…We plated keratinocytes onto a fibronectin-coated, elastic silicone gel coupled to glass. To quantify gel deformation due to cell-ECM traction force, we imaged fluorescent beads embedded in the silicone gel and measured the beads' displacements relative to their positions after removing the cells with proteinase K. We calculated inplane traction stresses, σ iz , from bead displacements and the substrate's elastic properties (38,39) …”

Section: Resultsmentioning

confidence: 99%

Cadherin-based intercellular adhesions organize epithelial cell–matrix traction forces

Mertz¹,

Che²,

Banerjee³

et al. 2012

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

217

201

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Cell-cell and cell-matrix adhesions play essential roles in the function of tissues. There is growing evidence for the importance of cross talk between these two adhesion types, yet little is known about the impact of these interactions on the mechanical coupling of cells to the extracellular matrix (ECM). Here, we combine experiment and theory to reveal how intercellular adhesions modulate forces transmitted to the ECM. In the absence of cadherin-based adhesions, primary mouse keratinocytes within a colony appear to act independently, with significant traction forces extending throughout the colony. In contrast, with strong cadherin-based adhesions, keratinocytes in a cohesive colony localize traction forces to the colony periphery. Through genetic or antibody-mediated loss of cadherin expression or function, we show that cadherin-based adhesions are essential for this mechanical cooperativity. A minimal physical model in which cell-cell adhesions modulate the physical cohesion between contractile cells is sufficient to recreate the spatial rearrangement of traction forces observed experimentally with varying strength of cadherin-based adhesions. This work defines the importance of cadherin-based cell-cell adhesions in coordinating mechanical activity of epithelial cells and has implications for the mechanical regulation of epithelial tissues during development, homeostasis, and disease. mechanotransduction | traction force microscopy M echanical interactions of individual cells have a crucial role in the spatial organization of tissues (1, 2) and in embryonic development (3-5). The mechanical cooperation of cells is evident in dynamic processes such as flow-induced alignment of vascular endothelial cells (6) and muscle contraction (7). However, mechanical interactions of cells within a tissue also affect the tissue's static mechanical properties including elastic modulus (8), surface tension (9), and fracture toughness (10). Little is known about how these tissue-scale mechanical phenomena emerge from interactions at the molecular and cellular levels (11).Tissue-scale mechanical phenomena are particularly important in developmental morphogenesis (12), homeostasis (13), and wound healing (14) in epithelial tissues. Cells exert mechanical force on each other at sites of intercellular adhesion, typically through cadherins (15, 16), as well as on the underlying extracellular matrix (ECM) through integrins (17-19). Cadherin-based adhesions can alter physical aspects of cells such as the surface tension of cellular aggregates (20) and the spreading (21) and migration (22) of single cells adherent to cadherin-patterned substrates. Integrity of intercellular adhesions may also contribute to metastatic potential (23). We and others have shown that epithelial cell clusters with strong cell-cell adhesions exhibit coordinated mechanical behavior over length scales much larger than a single cell (24-27). Several studies have implicated cross talk between cell-ECM and cell-cell adhesions (28, 29) that can be modulated by ac...

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Section: Resultsmentioning

confidence: 99%

Cadherin-based intercellular adhesions organize epithelial cell–matrix traction forces

Mertz¹,

Che²,

Banerjee³

et al. 2012

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

217

201

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“…Traction forces were analyzed using Fourier transform traction cytometry according to the methods discussed in (26)(27)(28). Fiducial motion was imaged using wide-field fluorescence microscopy, and particle displacement was calculated using particle-tracking velocimetry based on algorithms presented in (29).…”

Section: Traction Force Microscopymentioning

confidence: 99%

Frustrated Phagocytic Spreading of J774A-1 Macrophages Ends in Myosin II-Dependent Contraction

Kovari

Wei

Chang

et al. 2016

Biophysical Journal

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Conventional studies of dynamic phagocytic behavior have been limited in terms of spatial and temporal resolution due to the inherent three-dimensionality and small features of phagocytosis. To overcome these issues, we use a series of frustrated phagocytosis assays to quantitatively characterize phagocytic spreading dynamics. Our investigation reveals that frustrated phagocytic spreading occurs in phases and is punctuated by a distinct period of contraction. The spreading duration and peak contact areas are independent of the surface opsonin density, although the opsonin density does affect the likelihood that a cell will spread. This reinforces the idea that phagocytosis dynamics are primarily dictated by cytoskeletal activity. Structured illumination microscopy reveals that F-actin is reorganized during the course of frustrated phagocytosis. F-actin in early stages is consistent with that observed in lamellipodial protrusions. During the contraction phase, it is bundled into fibers that surround the cell and is reminiscent of a contractile belt. Using traction force microscopy, we show that cells exert significant strain on the underlying substrate during the contraction phase but little strain during the spreading phase, demonstrating that phagocytes actively constrict during late-stage phagocytosis. We also find that latestage contraction initiates after the cell surface area increases by 225%, which is consistent with the point at which cortical tension begins to rise. Moreover, reducing tension by exposing cells to hypertonic buffer shifts the onset of contraction to occur in larger contact areas. Together, these findings provide further evidence that tension plays a significant role in signaling late-stage phagocytic activity.

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“…These force measurements were strictly one-dimensional and were obtained either underneath a small portion of the foot using a force plate mounted vertically at an instant of time (Denny, 1980a) or integrated over a large area of the foot of the animal with a force plate mounted horizontally (Lissmann, 1945b). In our study, we obtained time series of high-resolution measurements of the spatial distribution of two-dimensional forces under the entire foot of the animal using a method that was initially developed in our laboratory to study the dynamics of cell migration (del Alamo et al, 2007). This traction cytometry method consists of measuring the deformation of a flat elastic substrate on which the animal is crawling.…”

Section: Dynamicsmentioning

confidence: 99%

The mechanics of the adhesive locomotion of terrestrial gastropods

Lai

Álamo

Rodríguez-Rodríguez

et al. 2010

Journal of Experimental Biology

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SUMMARYResearch on the adhesive locomotion of terrestrial gastropods is gaining renewed interest as it provides a source of guidance for the design of soft biomimetic robots that can perform functions currently not achievable by conventional rigid vehicles. The locomotion of terrestrial gastropods is driven by a train of periodic muscle contractions (pedal waves) and relaxations (interwaves) that propagate from their tails to their heads. These ventral waves interact with a thin layer of mucus secreted by the animal that transmits propulsive forces to the ground. The exact mechanism by which these propulsive forces are generated is still a matter of controversy. Specifically, the exact role played by the complex rheological and adhesive properties of the mucus is not clear. To provide quantitative data that could shed light on this question, we use a newly developed technique to measure, with high temporal and spatial resolution, the propulsive forces that terrestrial gastropods generate while crawling on smooth flat surfaces. The traction force measurements demonstrate the importance of the finite yield stress of the mucus in generating thrust and are consistent with the surface of the ventral foot being lifted with the passage of each pedal wave. We also show that a forward propulsive force is generated beneath each stationary interwave and that this net forward component is balanced by the resistance caused by the outer rim of the ventral foot, which slides at the speed of the center of mass of the animal. Simultaneously, the animal pulls the rim laterally inward. Analysis of the traction forces reveals that the kinematics of the pedal waves is far more complex than previously thought, showing significant spatial variation (acceleration/deceleration) as the waves move from the tail to the head of the animal. Supplementary material available online at

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Spatio-temporal analysis of eukaryotic cell motility by improved force cytometry

Cited by 195 publications

References 35 publications

Cadherin-based intercellular adhesions organize epithelial cell–matrix traction forces

Cadherin-based intercellular adhesions organize epithelial cell–matrix traction forces

Frustrated Phagocytic Spreading of J774A-1 Macrophages Ends in Myosin II-Dependent Contraction

The mechanics of the adhesive locomotion of terrestrial gastropods

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