Phenotype modulation in non-adherent and adherent sublines of Walker carcinosarcoma cells: the role of cell-substratum contacts and microtubules in controlling cell shape, locomotion and cytoskeletal structure

Sroka, Jolanta; Gunten, Michael von; Dunn, Graham; Keller, H. U.

doi:10.1016/s1357-2725(01)00178-9

Cited by 27 publications

(29 citation statements)

References 38 publications

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“…AdhSL cells could migrate efficiently on two-dimensional (2D) substrates ( Fig. 1D and Movie S1), as suggested by previous studies (16). In contrast, suspSL cells were nonadherent and displayed uncoordinated, random movements when placed on 2D substrates (Movie S2), but were able to migrate in confined environments, such as inside a micropipette or when placed between glass and agarose ( Fig.…”

Section: Sublines Of Walker Cells Can Form Either Lamellipodia or Blesupporting

confidence: 80%

“…Selection of the sublines was done as described in ref. 16, except that nonadherent suspSL cells were grown in ultra-low-attachment surface flasks (Corning Life Sciences) to prevent any substrate adhesion. Reagents and plasmids, cell preparation for microscopy, cell transfection, treatments, and microcontact printing are described in the SI Text.…”

Section: Methodsmentioning

confidence: 99%

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Cell mechanics control rapid transitions between blebs and lamellipodia during migration

Bergert

Chandradoss

Desai

et al. 2012

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

306

327

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Protrusion formation is an essential step during cell migration. Cells migrating in three-dimensional environments and in vivo can form a wide variety of protrusion types, including actin polymerizationdriven lamellipodia, and contractility-driven blebs. The ability to switch between different protrusions has been proposed to facilitate motility in complex environments and to promote cancer dissemination. However, plasticity in protrusion formation has so far mostly been investigated in the context of transitions between amoeboid and mesenchymal migration modes, which involve substantial changes in overall cell morphology. As a result, the minimal requirements of transitions between blebs and lamellipodia, as well as the time scales on which they occur, remain unknown. To address these questions, we investigated protrusion switching during cell migration at the single cell level. Using cells that can be induced to form either blebs or lamellipodia, we systematically assessed the mechanical requirements, as well as the dynamics, of switching between protrusion types. We demonstrate that shifting the balance between actin protrusivity and actomyosin contractility leads to immediate transitions between blebs and lamellipodia in migrating cells. Switching occurred without changes in global cell shape, polarity, or cell adhesion. Furthermore, rapid transitions between blebs and lamellipodia could also be triggered upon changes in substrate adhesion during migration on micropatterned surfaces. Together, our data reveal that the type of protrusion formed by migrating cells can be dynamically controlled independently of overall cell morphology, suggesting that protrusion formation is an autonomous module in the regulatory network that controls the plasticity of cell migration.

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Section: Sublines Of Walker Cells Can Form Either Lamellipodia or Blesupporting

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Cell mechanics control rapid transitions between blebs and lamellipodia during migration

Bergert

Chandradoss

Desai

et al. 2012

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

306

327

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“…Cells adopt different morphologies and modes of movement depending on various factors, such as adhesion (Friedl et al, 1998;Renkawitz et al, 2009;Sroka et al, 2002), contractility (Leader et al, 1983;Polte et al, 2004), Rho-family GTPase signalling (Kolyada et al, 2003;Sanz-Moreno et al, 2008) and the composition and rigidity of the extracellular matrix (ECM) (Pelham and Wang, 1997;Young and Herman, 1985). Changes in any of these factors can induce switches in cellular morphology and the mode of migration.…”

Section: Introductionmentioning

confidence: 99%

An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells

Lorentzen

Bamber

Sadok

et al. 2011

Journal of Cell Science

107

111

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SummaryMelanoma cells can switch between an elongated mesenchymal-type and a rounded amoeboid-type migration mode. The rounded 'amoeboid' form of cell movement is driven by actomyosin contractility resulting in membrane blebbing. Unlike elongated A375 melanoma cells, rounded A375 cells do not display any obvious morphological front-back polarisation, although polarisation is thought to be a prerequisite for cell movement. We show that blebbing A375 cells are polarised, with ezrin (a linker between the plasma membrane and actin cytoskeleton), F-actin, myosin light chain, plasma membrane, phosphatidylinositol (4,5)-bisphosphate and b1-integrin accumulating at the cell rear in a uropod-like structure. This structure does not have the typical protruding shape of classical leukocyte uropods, but, as for those structures, it is regulated by protein kinase C. We show that the ezrin-rich uropod-like structure (ERULS) is an inherent feature of polarised A375 cells and not a consequence of cell migration, and is necessary for cell invasion. Furthermore, we demonstrate that membrane blebbing is reduced at this site, leading to a model in which the rigid ezrin-containing structure determines the direction of a moving cell through localised inhibition of membrane blebbing. Journal of Cell ScienceIn confined spaces, blebbing cells can move forward without adhesion by a process termed 'chimneying' (Hawkins et al., 2009;Malawista and de Boisfleury Chevance, 1997). In fibrillar threedimensional matrices, such as collagen gels, blebs can expand into pores in the matrix, causing the squeezing phenotype observed in amoeboid lymphocytes and cancer cells in collagen gels (Haston and Shields, 1984;Wolf et al., 2003a). Cells that form small blebs can use them to 'elbow' their way through the meshwork of the ECM (Friedl and Wolf, 2009). The site of bleb formation determines the direction of cell movement (Keller and Bebie, 1996). However, some cells, such as A375 melanoma cells, form multiple blebs over the membrane and would therefore not be able to move directionally. However, A375 melanoma cells are able to invade into a collagen matrix and move in vitro and in vivo (Sahai and Marshall, 2003;Sanz-Moreno et al., 2008). As asymmetry is a prerequisite of cell movement (Petrie et al., 2009), this suggests some form of cellular polarisation. To understand cell movement driven by multiple blebs, we investigated polarisation in blebbing 'amoeboid' A375 melanoma cells. We show that A375 cells form an ezrin-rich uropod-like structure (which we call the ERULS) at their cell rear, and that this is required for invasion into a three-dimensional matrix. We present a model whereby accumulation of ezrin, membrane and actin in this structure reflects strong connections between the plasma membrane and the submembrane cortex, leading to the formation of a rigid structure that inhibits blebbing at the back of the cell. This leads to net movement of the cell in the direction opposite to the uropod-like structure. Our model explains how cells can move ...

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“…Mean Ϯ SD. initial proportion of polarised cells was relatively low, but not in cultures with a high proportion of polarised cells or in adhesive cells [Keller and Zimmermann, 1986;Sroka et al, 2002]. Most of the present experiments were performed with cultures showing a very high proportion of spontaneously polarised cells (Ͻ80%) and locomoting cells (Ͻ70%).…”

Section: Effects Of Colchicine And/or Latrunculin a On Locomotionmentioning

confidence: 99%

“…High concentrations of latrunculin A or cytochalasin suppress cell polarity and migration [Cassimeris et al, 1990;Keller and Zimmerman, 1986]. Microtubule-disassembling agents induce actomyosin-dependent contraction, they may uncover intrinsic polarity, stimulate contraction and/or migration, increase the F-actin content in neutrophils and/or nonadhesive Walker carcinosarcoma cells, and induce blebbing by dissociation of the plasma membrane from the actin cortex [Danowski, 1989;Keller et al, 1984;Keller and Zimmermann, 1986;Keller and Eggli, 1998;Sroka et al, 2002;Keller et al, 2002]. Substantial evidence for positive feedback interactions between microtubules and actin dynamics in cell motility has emerged [Singer and Kupfer,1986;Waterman-Storer and Salmon, 1999;Small et al, 1999].…”

Section: Introductionmentioning

confidence: 99%

Localised depletion of polymerised actin at the front of Walker carcinosarcoma cells increases the speed of locomotion

Keller

Zadeh

Eggli

2002

Cell Motil. Cytoskeleton

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Spontaneously migrating Walker carcinosarcoma cells usually form lamellipodia at the front. Combined treatment with 10(-5)M colchicine and 10(-7)M latrunculin A produces large defects in the cortical F-actin layer at the leading front and suppresses lamellipodia. However, the cortical actin layer at the rear is intact and shows myosin IIA accumulation. These cells, showing no or little detectable cortical F-actin at the front and no morphologically recognisable protrusions, migrate faster than control cells with lamellipodia and an intact cortical actin layer. This documents that the cortical actin layer or actin-powered force generation at the front is redundant for locomotion. Colchicine and latrunculin A have synergistic effects in compromising the cortical layer at the front and in increasing the speed of locomotion, but antagonistic effects on the relative amount of F-actin per cell. Colchicine but not latrunculin A, can increase the proportion of polarised and locomoting cells under appropriate conditions. Locomotion and polarity of cells treated with latrunculin A and colchicine is inhibited at latrunculin A concentrations >10(-7)M, by the myosin inhibitor BDM or the ROCK inhibitor Y-27632. Colchicine and Y-27632 have antagonistic effects on polarity and the speed of locomoting cells. The data show that locomotion of metazoan cells, which normally form lamellipodia, can be driven by actomyosin contraction behind the front (cell body, uropod). They are best compatible with a cortical contraction/frontal expansion model, but they are not compatible with models implying that actin polymerisation or actomyosin contraction at the front drive locomotion of the cells studied.

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Phenotype modulation in non-adherent and adherent sublines of Walker carcinosarcoma cells: the role of cell-substratum contacts and microtubules in controlling cell shape, locomotion and cytoskeletal structure

Cited by 27 publications

References 38 publications

Cell mechanics control rapid transitions between blebs and lamellipodia during migration

Cell mechanics control rapid transitions between blebs and lamellipodia during migration

An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells

Localised depletion of polymerised actin at the front of Walker carcinosarcoma cells increases the speed of locomotion

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