Rho GTPase signaling modulates cell shape and contractile phenotype in an isoactin-specific manner

Kolyada, Alexey; Riley, Kathleen N.; Herman, Ira M.

doi:10.1152/ajpcell.00177.2003

Cited by 39 publications

(66 citation statements)

References 31 publications

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“…23,24 Importantly, a recent role for Rho GTPase-dependent signal transduction has now been suggested in the control of pericyte shape and contractility, leading to microvascular tone and blood flow regulation via alterations in cytoskeletal dynamics. 25 In this study, we demonstrate that pericyte-specific and Rho GTPase-dependent signal transduction reversibly regulates both pericyte contractility and capillary endothelial cell growth state. Building on the previous work of Harris et al 26 and others, we have developed two novel in vitro systems to directly quantify and simultaneously link the contractile potential of microvascular pericytes with pericyte Rho GTPase-mediated endothelial cell growth control.…”

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confidence: 54%

Pericyte Rho GTPase Mediates Both Pericyte Contractile Phenotype and Capillary Endothelial Growth State

Kutcher

Kolyada

Surks

et al. 2007

The American Journal of Pathology

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117

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Pericytes regulate microvascular development and maturation through the control of endothelial cell motility, proliferation, and differentiation. The Rho GTPases have recently been described as key regulators of pericyte shape and contractile phenotype by signaling through the actin cytoskeleton in an isoactin-specific manner. In this report, we reveal that Rho GTPase-dependent signal transduction not only influences pericyte shape and contractile potential but also modulates capillary endothelial proliferative status and pericyte-endothelial interactions in vitro. We provide evidence that overexpression of mutant Rho GTPases, but not other Ras-related small GTPases, significantly alters pericyte shape, contractility, and endothelial growth state in microvascular cell co-cultures. In particular, we describe the use of a silicon substrate deformation assay to demonstrate that pericyte contractility is Rho GTP-and Rho kinase-dependent; further, we describe a novel in vitro system for examining pericyte-mediated endothelial growth arrest and show that control pericytes are capable of growtharresting capillary endothelial cells in a cell contactdependent manner, whereas pericytes overexpressing dominant-active and -negative Rho GTPase are comparably incompetent. These data strongly suggest that signaling through the pericyte Rho GTPase pathway may provide critical cues to the processes of microvascular stabilization, maturation, and contractility during development and disease.

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confidence: 54%

Pericyte Rho GTPase Mediates Both Pericyte Contractile Phenotype and Capillary Endothelial Growth State

Kutcher

Kolyada

Surks

et al. 2007

The American Journal of Pathology

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117

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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

108

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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“…Furthermore, specific cytoskeletondisrupting agents/pharmacological inhibitors similarly reverse the enhanced cytoskeletal stiffening and contractile activity (41). Reorganization/expression of smooth muscle proteins underlies contractile force generation and has been shown to be dependent on Rho signaling in a variety of cell types (32,35,39,44,48,68). Moreover, ROCK can directly induce smooth muscle contraction via phosphorylation of MLC (37) and has been implicated in the generation of central stress fibers via increased phosphorylated MLC levels, while MLCK plays a role in assembly of peripheral stress fibers (31, 69).…”

Section: Mrip Silencing In Pericytesmentioning

confidence: 99%

“…Similar to MRIP-silenced, hypertrophic pericytes that are unable to sustain CEC growth arrest in a contact-dependent manner, pericytes with altered Rho GTP status or ROCK activity influence both pericyte contractile phenotype and abrogate maintenance of EC growth arrest (35,36,39,41). While S phase entry is not always indicative of cytokinesis, preliminary data reveal an increase in the ratio of CEC to pericytes and in the total number of EC in MRIP-silenced cocultures (1.7-and 1.4-fold, respectively; data not shown) on day 7.…”

Section: Coculture Of Cec and Mrip-silenced Pericytesmentioning

confidence: 99%

“…Overexpression and selective inhibition studies have revealed that RhoA GTPase activity affects specifically the smooth muscle actin (SMA)-based cytoskeletal networks (35). Furthermore, alteration of pericyte Rho GTPase status via viral transduction of dominantactive or dominant-negative RhoA constructs leads to altered force generation on deformable substrates and hyper-or hypocontractile cells, respectively (39).…”

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confidence: 99%

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Pericyte contractility controls endothelial cell cycle progression and sprouting: insights into angiogenic switch mechanics

Durham

Surks

Dulmovits

et al. 2014

American Journal of Physiology-Cell Physiology

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Durham JT, Surks HK, Dulmovits BM, Herman IM. Pericyte contractility controls endothelial cell cycle progression and sprouting: insights into angiogenic switch mechanics. Am J Physiol Cell Physiol 307: C878 -C892, 2014. First published August 20, 2014; doi:10.1152/ajpcell.00185.2014.-Microvascular stability and regulation of capillary tonus are regulated by pericytes and their interactions with endothelial cells (EC). While the RhoA/Rho kinase (ROCK) pathway has been implicated in modulation of pericyte contractility, in part via regulation of the myosin light chain phosphatase (MLCP), the mechanisms linking Rho GTPase activity with actomyosin-based contraction and the cytoskeleton are equivocal. Recently, the myosin phosphatase-RhoA-interacting protein (MRIP) was shown to mediate the RhoA/ROCK-directed MLCP inactivation in vascular smooth muscle. Here we report that MRIP directly interacts with the ␤-actinspecific capping protein ␤cap73. Furthermore, manipulation of MRIP expression influences pericyte contractility, with MRIP silencing inducing cytoskeletal remodeling and cellular hypertrophy. MRIP knockdown induces a repositioning of ␤cap73 from the leading edge to stress fibers; thus MRIP-silenced pericytes increase F-actin-driven cell spreading twofold. These hypertrophied and cytoskeleton-enriched pericytes demonstrate a 2.2-fold increase in contractility upon MRIP knockdown when cells are plated on a deformable substrate. In turn, silencing pericyte MRIP significantly affects EC cycle progression and angiogenic activation. When MRIP-silenced pericytes are cocultured with capillary EC, there is a 2.0-fold increase in EC cycle entry. Furthermore, in three-dimensional models of injury and repair, silencing pericyte MRIP results in a 1.6-fold elevation of total tube area due to EC network formation and increased angiogenic sprouting. The pivotal role of MRIP expression in governing pericyte contractile phenotype and endothelial growth should lend important new insights into how chemomechanical signaling pathways control the "angiogenic switch" and pathological angiogenic induction. cytoskeleton; Rho kinase; myosin light chain phosphatase; myosin light chain; capillary; mural cell; angiogenesis IN ARTERIES AND VEINS, vascular tone is largely regulated via vascular smooth muscle. In capillaries and venules, however, tonus is coregulated by endothelial cells (EC) and pericytes. In the microcirculation, pericyte dysfunction contributes to several debilitating vascular pathologies, including diabetic retinopathy (DR), hypertension, atherosclerosis, ischemia, stroke, asthma, and Alzheimer's disease (10,13,30,53).A growing body of evidence in pericyte pathophysiology indicates that pericytes play a critical role in the maintenance of capillary EC (CEC) physiology and angiogenic induction (2,12,16,38). For example, in the retina, perturbations in pericyte-EC interactions can result in pathological angiogenesis accompanying proliferative DR (2, 13, 21). The notion has been widely held that, early in DR, vascular cell ...

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Rho GTPase signaling modulates cell shape and contractile phenotype in an isoactin-specific manner

Cited by 39 publications

References 31 publications

Pericyte Rho GTPase Mediates Both Pericyte Contractile Phenotype and Capillary Endothelial Growth State

Pericyte Rho GTPase Mediates Both Pericyte Contractile Phenotype and Capillary Endothelial Growth State

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

Pericyte contractility controls endothelial cell cycle progression and sprouting: insights into angiogenic switch mechanics

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