scholarly journals Nonmuscle Myosin IIB Is Involved in the Guidance of Fibroblast Migration

2004 ◽  
Vol 15 (3) ◽  
pp. 982-989 ◽  
Author(s):  
Chun-Min Lo ◽  
Denis B. Buxton ◽  
Gregory C.H. Chua ◽  
Micah Dembo ◽  
Robert S. Adelstein ◽  
...  
Keyword(s):  
Cell Migration ◽  
Large Fraction ◽  
Myosin Ii ◽  
Cell Movement ◽  
Traction Forces ◽  
Migration Patterns ◽  
Fibroblast Migration ◽  
Substrate Rigidity ◽  
Migration Analysis ◽  
Mutant Cells

Although myosin II is known to play an important role in cell migration, little is known about its specific functions. We have addressed the function of one of the isoforms of myosin II, myosin IIB, by analyzing the movement and mechanical characteristics of fibroblasts where this protein has been ablated by gene disruption. Myosin IIB null cells displayed multiple unstable and disorganized protrusions, although they were still able to generate a large fraction of traction forces when cultured on flexible polyacrylamide substrates. However, the traction forces were highly disorganized relative to the direction of cell migration. Analysis of cell migration patterns indicated an increase in speed and decrease in persistence, which were likely responsible for the defects in directional movements as demonstrated with Boyden chambers. In addition, unlike control cells, mutant cells failed to respond to mechanical signals such as compressing forces and changes in substrate rigidity. Immunofluorescence staining indicated that myosin IIB was localized preferentially along stress fibers in the interior region of the cell. Our results suggest that myosin IIB is involved not in propelling but in directing the cell movement, by coordinating protrusive activities and stabilizing the cell polarity.

The Dictyostelium RasS protein is required for macropinocytosis, phagocytosis and the control of cell movement

2000 ◽  
Vol 113 (4) ◽  
pp. 709-719 ◽  
Author(s):  
J.R. Chubb ◽  
A. Wilkins ◽  
G.M. Thomas ◽  
R.H. Insall
Keyword(s):  
Cell Migration ◽  
Significant Proportion ◽  
Cell Movement ◽  
Fluid Phase ◽  
Small Gtpases ◽  
Cell Cortex ◽  
Fluid Phase Endocytosis ◽  
Ras Family ◽  
Actin Protrusions ◽  
Mutant Cells

Endocytosis and cell migration both require transient localised remodelling of the cell cortex. Several lines of evidence suggest a key regulatory role in these activities for members of the Ras family of small GTPases. We have generated Dictyostelium cells lacking one member of this family, RasS, and the mutant cells are perturbed in endocytosis and cell migration. Mutant amoebae are defective in phagocytosis and fluid-phase endocytosis and are impaired in growth. Conversely, the rasS(-)cells show an enhanced rate of cell migration, moving three times faster than wild-type controls. The mutant cells display an aberrant morphology, are highly polarised, carry many elongated actin protrusions and show a concomitant decrease in formation of pinocytic crowns on the cell surface. These morphological aberrations are paralleled by changes in the actin cytoskeleton, with a significant proportion of the cortical F-actin relocalised to prominent pseudopodia. Rapid migration and endocytosis appear to be mutually incompatible and it is likely that RasS protein is required to maintain the normal balance between these two actin-dependent processes.

scholarly journals αvβ3 and α5β1 integrin recycling pathways dictate downstream Rho kinase signaling to regulate persistent cell migration

2007 ◽  
Vol 177 (3) ◽  
pp. 515-525 ◽  
Author(s):  
Dominic P. White ◽  
Patrick T. Caswell ◽  
Jim C. Norman
Keyword(s):  
Cell Migration ◽  
Rho Kinase ◽  
Cell Movement ◽  
Αvβ3 Integrin ◽  
Protein Kinase D1 ◽  
Fibroblast Migration ◽  
Short Loop ◽  
Cell Front ◽  
Α5β1 Integrin ◽  
Persistent Cell

Accumulating evidence suggests that integrin recycling regulates cell migration. However, the lack of reagents to selectively target the trafficking of individual heterodimers, as opposed to endocytic transport as a whole, has made it difficult to define the contribution made by particular recycling pathways to directional cell movement. We show that autophosphorylation of protein kinase D1 (PKD1) at Ser916 is necessary for its association with αvβ3 integrin. Expression of PKD1916A or the use of mutants of β3 that do not bind to PKD1 selectively inhibits short-loop, Rab4-dependent recycling of αvβ3, and this suppresses the persistence of fibroblast migration. However, we report that short-loop recycling does not directly contribute to fibroblast migration by moving αvβ3 to the cell front, but by antagonizing α5β1 recycling, which, in turn, influences the cell's decision to migrate with persistence or to move randomly.

scholarly journals Robust Quantification of Regional Patterns of Migration in Three-Dimensional Cell Culture Models

2021 ◽  
Author(s):  
Chun Kiet Vong ◽  
Alan Wang ◽  
Mike Dragunow ◽  
Thomas I-H Park ◽  
Vickie Shim
Keyword(s):  
Cell Migration ◽  
Three Dimensional ◽  
Regional Patterns ◽  
Migration Patterns ◽  
Regional Migration ◽  
Migration Model ◽  
Culture Models ◽  
Drug Treatments ◽  
Migration Analysis

Abstract Wound healing assays is a common two-dimensional migration model, with the spheroid assay three-dimensional migration model recently emerging as being more representative of in vivo migration behaviours. These models provide insight to the overall migration of cells in response to various factors such as biological, chemotactic and molecular agents. However, currently available analysis techniques for these assays fall short on providing quantifiable means to measure regional migration patterns, , which is essential to allow more robust assessment of drug treatments on cell migration in a chemotactic fashion. Therefore, the aim of this study is to develop a finite element (FE) based pipeline that can objectively quantify regional migration patterns of cells. Here, we report that our FE based approach was able to accurately measure changes in overall migration areas compared to the standard ImageJ method. Furthermore, our regional migration analysis provided accurate and quantitative means to analyse the migration pattern seen in the phantom data and our experimental results, giving us confidence that it can be a robust tool for analysing cell migration patterns.

scholarly journals A Cellular Potts Model for Analyzing Cell Migration across Constraining Pillar Arrays

Axioms ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 32
Author(s):  
Marco Scianna ◽  
Luigi Preziosi
Keyword(s):  
Cell Migration ◽  
Potts Model ◽  
Large Fraction ◽  
Cell Movement ◽  
Stochastic Approach ◽  
Cellular Potts Model ◽  
Cell Dynamics ◽  
Experimental Systems ◽  
Pillar Arrays ◽  
Constrained Environments

Cell migration in highly constrained environments is fundamental in a wide variety of physiological and pathological phenomena. In particular, it has been experimentally shown that the migratory capacity of most cell lines depends on their ability to transmigrate through narrow constrictions, which in turn relies on their deformation capacity. In this respect, the nucleus, which occupies a large fraction of the cell volume and is substantially stiffer than the surrounding cytoplasm, imposes a major obstacle. This aspect has also been investigated with the use of microfluidic devices formed by dozens of arrays of aligned polymeric pillars that limit the available space for cell movement. Such experimental systems, in particular, in the designs developed by the groups of Denais and of Davidson, were here reproduced with a tailored version of the Cellular Potts model, a grid-based stochastic approach where cell dynamics are established by a Metropolis algorithm for energy minimization. The proposed model allowed quantitatively analyzing selected cell migratory determinants (e.g., the cell and nuclear speed and deformation, and forces acting at the nuclear membrane) in the case of different experimental setups. Most of the numerical results show a remarkable agreement with the corresponding empirical data.

scholarly journals Distinct Roles of Frontal and Rear Cell-Substrate Adhesions in Fibroblast Migration

2001 ◽  
Vol 12 (12) ◽  
pp. 3947-3954 ◽  
Author(s):  
Steven Munevar ◽  
Yu-li Wang ◽  
Micah Dembo
Keyword(s):  
Cell Migration ◽  
Traction Force ◽  
Leading Edge ◽  
Traction Forces ◽  
Passive Resistance ◽  
Fibroblast Migration ◽  
3T3 Fibroblasts ◽  
Cell Substrate ◽  
Extensive Cell ◽  
Physical And Chemical

Cell migration involves complex physical and chemical interactions with the substrate. To probe the mechanical interactions under different regions of migrating 3T3 fibroblasts, we have disrupted cell-substrate adhesions by local application of the GRGDTP peptide, while imaging stress distribution on the substrate with traction force microscopy. Both spontaneous and GRGDTP-induced detachment of the trailing edge caused extensive cell shortening, without changing the overall level of traction forces or the direction of migration. In contrast, disruption of frontal adhesions caused dramatic, global loss of traction forces before any significant shortening of the cell. Although traction forces and cell migration recovered within 10–20 min of transient frontal treatment, persistent treatment with GRGDTP caused the cell to develop traction forces elsewhere and reorient toward a new direction. We conclude that contractile forces of a fibroblast are transmitted to the substrate through two distinct types of adhesions. Leading edge adhesions are unique in their ability to transmit active propulsive forces. Their functions cannot be transferred directly to existing adhesions upon detachment. Trailing end adhesions create passive resistance during cell migration and readily redistribute their loads upon detachment. Our results indicate the distinct nature of mechanical interactions at the leading versus trailing edges, which together generate the mechanical interactions for fibroblast migration.

Traction Force Measurement During Collective Cell Migration Measured by Multichannel Micropillar Device

2013 ◽  
Author(s):  
Toshiro Ohashi ◽  
Akito Sugawara
Keyword(s):  
Cell Migration ◽  
Soft Lithography ◽  
Force Measurement ◽  
Single Cells ◽  
Cell Movement ◽  
Traction Force ◽  
Collective Cell Migration ◽  
Maximum Magnitude ◽  
Traction Forces ◽  
Moving Front

Cell migration is essential for a variety of biological and pathological processes such as wound healing, inflammation and tumor metastasis. However, the mechanical environment within a group of cells during collective migration has not been well characterized. In this study, a polydimethylsiloxane (PDMS) multichannel device was fabricated using standard photolithography and soft lithography techniques and was used to monitor cellular traction forces during migration. A migration rate of 5.7 μm/h was measured in microchannels and leading cells in the moving front of the migration generated traction forces with a maximum magnitude of 14 nN at their front side. Traction forces generated by cells behind the leading cells directed forces backward at both the front and rear sides. However, traction forces generated by cells behind the second row had forces in random directions and with smaller magnitudes compared to those on the front and the second row. It is assumed that cells on the front line generated large traction forces and migrated actively as single cells, pulling adjacent cells forward, whereas the cell movement after the third row was restricted by mechanical linkages between their neighboring cells.

scholarly journals A three-component mechanism for fibroblast migration with a contractile cell body that couples a myosin II–independent propulsive anterior to a myosin II–dependent resistive tail

2012 ◽  
Vol 23 (9) ◽  
pp. 1657-1663 ◽  
Author(s):  
Wei-hui Guo ◽  
Yu-li Wang
Keyword(s):  
Cell Migration ◽  
Cell Body ◽  
Nuclear Translocation ◽  
Myosin Ii ◽  
Local Treatment ◽  
Anterior Region ◽  
Nuclear Movement ◽  
Forward Movement ◽  
Cultured Fibroblasts ◽  
Fibroblast Migration

To understand the mechanism of cell migration, we cultured fibroblasts on micropatterned tracks to induce persistent migration with a highly elongated morphology and well-defined polarity, which allows microfluidic pharmacological manipulations of regional functions. The function of myosin II was probed by applying inhibitors either globally or locally. Of interest, although global inhibition of myosin II inhibited tail retraction and caused dramatic elongation of the posterior region, localized inhibition of the cell body inhibited nuclear translocation and caused elongation of the anterior region. In addition, local application of cytochalasin D at the tip inhibited frontal extension without inhibiting forward movement of the cell nucleus, whereas local treatment posterior to the nucleus caused reversal of nuclear movement. Imaging of cortical dynamics indicated that the region around the nucleus is a distinct compression zone where activities of anterior and posterior regions converge. These observations suggest a three-component model of cell migration in which a contractile middle section is responsible for the movement of a bulky cell body and the detachment/retraction of a resistive tail, thereby allowing these regions to undergo coordinated movement with a moving anterior region that carries little load.

scholarly journals Usiigaci: Instance-aware cell tracking in stain-free phase contrast microscopy enabled by machine learning

2019 ◽  
Author(s):  
Hsieh-Fu Tsai ◽  
Joanna Gajda ◽  
Tyler F.W. Sloan ◽  
Andrei Rares ◽  
Amy Q. Shen
Keyword(s):  
Cell Migration ◽  
Phase Contrast ◽  
Cell Tracking ◽  
Morphological Changes ◽  
Cell Movement ◽  
Cell Segmentation ◽  
Phase Contrast Microscopy ◽  
3T3 Fibroblasts ◽  
Contrast Microscopy ◽  
Migration Analysis

AbstractStain-free, single-cell segmentation and tracking is tantamount to the holy grail of microscopic cell migration analysis. Phase contrast microscopy (PCM) images with cells at high density are notoriously difficult to segment accurately; thus, manual segmentation remains the de facto standard practice. In this work, we introduce Usiigaci, an all-in-one, semi-automated pipeline to segment, track, and visualize cell movement and morphological changes in PCM. Stain-free, instance-aware segmentation is accomplished using a mask regional convolutional neural network (Mask R-CNN). A Trackpy-based cell tracker with a graphical user interface is developed for cell tracking and data verification. The performance of Usiigaci is validated with electrotaxis of NIH/3T3 fibroblasts. Usiigaci provides highly accurate cell movement and morphological information for quantitative cell migration analysis.

Protein tyrosine phosphatase-α complexes with the IGF-I receptor and undergoes IGF-I-stimulated tyrosine phosphorylation that mediates cell migration

2009 ◽  
Vol 297 (1) ◽  
pp. C133-C139 ◽  
Author(s):  
Shirley C. Chen ◽  
Ranvikram S. Khanna ◽  
Darrell C. Bessette ◽  
Lionel A. Samayawardhena ◽  
Catherine J. Pallen
Keyword(s):  
Cell Migration ◽  
Tyrosine Phosphorylation ◽  
Protein Tyrosine Phosphatase ◽  
Tyrosine Kinases ◽  
Tyrosine Phosphatase ◽  
Phosphorylation Site ◽  
Protein Tyrosine ◽  
Fibroblast Migration ◽  
Igf I ◽  
In Cells

Protein tyrosine phosphatase-α (PTPα) is a widely expressed receptor-type phosphatase that functions in multiple signaling systems. The actions of PTPα can be regulated by its phosphorylation on serine and tyrosine residues, although little is known about the conditions that promote PTPα phosphorylation. In this study, we tested the ability of several extracellular factors to stimulate PTPα tyrosine phosphorylation. The growth factors IGF-I and acidic FGF induced the highest increase in PTPα phosphorylation at tyrosine 789, followed by PMA and lysophosphatidic acid, while EGF had little effect. Further investigation of IGF-I-induced PTPα tyrosine phosphorylation demonstrated that this occurs through a novel Src family kinase-independent mechanism that does not require focal adhesion kinase, phosphatidylinositol 3-kinase, or MEK. We also show that PTPα physically interacts with the IGF-I receptor. In contrast to IGF-I-induced PTPα phosphorylation, this association does not require IGF-I. The interaction of PTPα and the IGF-I receptor is independent of PTPα catalytic activity, and expression of exogenous PTPα does not promote IGF-I receptor tyrosine dephosphorylation, indicating that PTPα does not act as an IGF-I receptor phosphatase. However, PTPα mediates IGF-I signaling, because IGF-I-stimulated fibroblast migration was reduced by ∼50% in cells lacking PTPα or in cells with mutant PTPα lacking the tyrosine 789 phosphorylation site. Our results suggest that PTPα tyrosine phosphorylation can occur in response to diverse stimuli and can be mediated by various tyrosine kinases. In the case of IGF-I, we propose that IGF-I-induced tyrosine 789 phosphorylation of PTPα, possibly catalyzed by the PTPα-associated IGF-I receptor tyrosine kinase, is required for efficient cell migration in response to this growth factor.

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