AXIAL-SYMMETRIC MODELING AND KINEMATIC ANALYSIS OF SPREADING OF SPARSELY CULTURED FIBROBLASTS

Cell spreading plays an important role in the modulation of physiological functions such as inflammation and cancer metastasis. The Brownian ratchet model and Bell's model have been used to simulate actin dynamics and bond kinetics for focal adhesion dynamics, respectively. In the present study, these models were modified and two additional subcellular mechanisms, integrin and myosin kinetics, were incoporated. An integrin recruitment function was introduced to determine the size of a focal adhesion associated with the substrate stiffness. The relationship between myosin concentration and the actin protrusion velocity was described by a first-order differential equation. Subcellular processes, including cell protrusion, focal adhesion formation, and stress fiber formation, were integrated into an axial-symmetric biophysical model, while inputs to the model were kinematic data from time-lapse experiments. Numerical simulations of the model using the Gillespie algorithm showed that dynamics of cell spreading can be well described by the model.

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Regulation of the RhoA pathway in human endothelial cell spreading on type IV collagen: role of calcium influx

Journal of Cell Science ◽

10.1242/jcs.112.19.3205 ◽

1999 ◽

Vol 112 (19) ◽

pp. 3205-3213 ◽

Cited By ~ 1

Author(s):

L. Masiero ◽

K.A. Lapidos ◽

I. Ambudkar ◽

E.C. Kohn

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Focal Adhesion ◽

Type Iv Collagen ◽

Cell Spreading ◽

Stress Fiber ◽

Fiber Formation ◽

Stress Fibers ◽

Type Iv ◽

Stress Fiber Formation

We have shown that nonvoltage-operated Ca(2+) entry regulates human umbilical vein endothelial cell adhesion, migration, and proliferation on type IV collagen. We now demonstrate a requirement for Ca(2+) influx for activation of the RhoA pathway during endothelial cell spreading on type IV collagen. Reorganization of actin into stress fibers was complete when the cells where fully spread at 90 minutes. No actin organization into stress fibers was seen in endothelial cells plated on type I collagen, indicating a permissive effect of type IV collagen. CAI, a blocker of nonvoltage-operated Ca(2+) channels, prevented development of stress fiber formation in endothelial cells on type IV collagen. This permissive effect was augmented by Ca(2+) influx, as stimulated by 0. 5 microM thapsigargin or 0.1 microM ionomycin, yielding faster development of actin stress fibers. Ca(2+) influx and actin rearrangement in response to thapsigargin and ionomycin were abrogated by CAI. Activated, membrane-bound RhoA is a substrate for C3 exoenzyme which ADP-ribosylates and inactivates RhoA, preventing actin stress fiber formation. Pretreatment of endothelial cells with C3 exoenzyme prevented basal and thapsigargin-augmented stress fiber formation. While regulation of Ca(2+) influx did not alter RhoA translocation, it reduced in vitro ADP-ribosylation of RhoA (P(2)<0. 05), suggesting Ca(2+) influx is needed for RhoA activation during spreading on type IV collagen; no Ca(2+) regulated change in RhoA was seen in HUVECs spreading on type I collagen matrix. Blockade of Ca(2+) influx of HUVEC spread on type IV collagen also reduced tyrosine phosphorylation of p190Rho-GAP and blocked thapsigargin-enhanced binding of p190Rho-GAP to focal adhesion kinase. Thus, Ca(2+) influx is necessary for RhoA activation and for linkage of the RhoA/stress fiber cascade to the focal adhesion/focal adhesion kinase pathway during human umbilical vein endothelial cell spreading on type IV collagen.

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Nanoporosity Stimulates Cell Spreading and Focal Adhesion Formation in Cells with Mutated Paxillin

ACS Applied Materials & Interfaces ◽

10.1021/acsami.0c01172 ◽

2020 ◽

Vol 12 (13) ◽

pp. 14924-14932

Author(s):

Dainelys Guadarrama Bello ◽

Aurélien Fouillen ◽

Antonella Badia ◽

Antonio Nanci

Keyword(s):

Focal Adhesion ◽

Adhesion Formation ◽

Cell Spreading ◽

Focal Adhesion Formation ◽

In Cells

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Rap1 Can Bypass the FAK-Src-Paxillin Cascade to Induce Cell Spreading and Focal Adhesion Formation

PLoS ONE ◽

10.1371/journal.pone.0050072 ◽

2012 ◽

Vol 7 (11) ◽

pp. e50072 ◽

Cited By ~ 11

Author(s):

Sarah H. Ross ◽

Emma Spanjaard ◽

Anneke Post ◽

Marjolein J. Vliem ◽

Hendy Kristyanto ◽

...

Keyword(s):

Focal Adhesion ◽

Adhesion Formation ◽

Cell Spreading ◽

Focal Adhesion Formation

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Alteration of transbilayer phospholipid compositions is involved in cell adhesion, cell spreading, and focal adhesion formation

FEBS Letters ◽

10.1002/1873-3468.12247 ◽

2016 ◽

Vol 590 (14) ◽

pp. 2138-2145 ◽

Cited By ~ 7

Author(s):

Rie Miyano ◽

Takashi Matsumoto ◽

Hiroyuki Takatsu ◽

Kazuhisa Nakayama ◽

Hye-Won Shin

Keyword(s):

Cell Adhesion ◽

Focal Adhesion ◽

Adhesion Formation ◽

Cell Spreading ◽

Focal Adhesion Formation

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p130Cas contributes to cellular mechanosensing and force exertion

10.1101/371021 ◽

2018 ◽

Author(s):

Hedde van Hoorn ◽

Dominique M. Donato ◽

H. Emrah Balcioglu ◽

Erik H. Danen ◽

Thomas Schmidt

Keyword(s):

Focal Adhesion ◽

Adhesion Formation ◽

Focal Adhesions ◽

Full Length ◽

Actin Dynamics ◽

Knock Out ◽

Focal Adhesion Formation ◽

Pillar Arrays ◽

Cellular Machinery ◽

And Migration

AbstractCell survival, differentiation, and migration are all dependent on the cell’s interaction with its external environment. In addition to chemical cues, cells react to their physical environment, particularly the stiffness of the substrate. In order for cells to react to these elements, they must make use of cellular machinery to signal changes in their microenvironment. One such proposed machinery is the protein p130Cas, which has been shown to regulate focal adhesion turnover, actin dynamics, and cell migration. Here we show that p130Cas localizes to focal adhesions depending on substrate stiffness and subsequently modulates cellular force exertion. We compared on substrates of tunable stiffness knock-out CAS-/-cells to cells re-expressing either the full-length p130Cas or a mutant lacking the focal adhesion targeting domains. On polyacrylamide gels, we observed that p130Cas prevented focal adhesion formation at low stiffness. On structured micro-pillar arrays, p130Cas preferentially localized to sites of force exertion when the apparent Young’s modulus of the substrate was higher than E = 47 kPa. Stiffness-dependent localization of p130Cas coincided with slower, but increased force exertion for the full-length p130Cas. Cas localization to focal adhesions preceded force build-up by three minutes, suggesting a coordinating role for p130Cas in the cellular mechanoresponse. Thus, p130Cas appears to relay mechanosensory information in the cell through its ability to tune force exertion at the focal adhesion.

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Activation of Rho-kinase and focal adhesion kinase regulates the organization of stress fibers and focal adhesions in the central part of fibroblasts

10.7287/peerj.preprints.3297v1 ◽

2017 ◽

Author(s):

Kazuo Katoh

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Adhesion Formation ◽

Rho Kinase ◽

Focal Adhesions ◽

Fiber Formation ◽

Stress Fibers ◽

Focal Adhesion Formation ◽

Specific Regulation

Specific regulation and activation of focal adhesion kinase (FAK) are thought to be important for focal adhesion formation, and activation of Rho-kinase has been suggested to play a role in determining the effects of FAK on the formation of stress fibers and focal adhesions. To clarify the role of FAK in stress fiber formation and focal adhesion organization, we examined the formation of new stress fibers and focal adhesions by activation of Rho-kinase in FAK knockout (FAK–/–) fibroblasts. FAK–/– cells were elliptical in shape, and showed reduced numbers of stress fibers and focal adhesions in the central part of the cells along with large focal adhesions in the peripheral regions. Activation of Rho-kinase in FAK–/– cells transiently increased the actin filaments in the cell center, but these did not form typical thick stress fibers. Moreover, only plaque-like structures as the origins of newly formed focal adhesions were observed in the center of the cell. Furthermore, introduction of an exogenous GFP-labeled FAK gene into FAK–/– cells resulted in increased numbers of stress fibers and focal adhesions in the center of the cells, which showed typical fibroblast morphology. These results indicated that FAK plays an important role in the formation of stress fibers and focal adhesions as well as in regulation of cell shape and morphology with the activation of Rho-kinase.

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Smooth muscle cell rigidity and extracellular matrix organization influence endothelial cell spreading and adhesion formation in coculture

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00618.2007 ◽

2007 ◽

Vol 293 (3) ◽

pp. H1978-H1986 ◽

Cited By ~ 22

Author(s):

Charles S. Wallace ◽

Sophie A. Strike ◽

George A. Truskey

Keyword(s):

Extracellular Matrix ◽

Smooth Muscle ◽

Endothelial Cell ◽

Focal Adhesion ◽

Adhesion Formation ◽

Focal Adhesions ◽

Cell Spreading ◽

Tissue Engineered Blood Vessels ◽

Focal Adhesion Formation ◽

Fibrillar Adhesion

Efforts to develop functional tissue-engineered blood vessels have focused on improving the strength and mechanical properties of the vessel wall, while the functional status of the endothelium within these vessels has received less attention. Endothelial cell (EC) function is influenced by interactions between its basal surface and the underlying extracellular matrix. In this study, we utilized a coculture model of a tissue-engineered blood vessel to evaluate EC attachment, spreading, and adhesion formation to the extracellular matrix on the surface of quiescent smooth muscle cells (SMCs). ECs attached to and spread on SMCs primarily through the α5β1-integrin complex, whereas ECs used either α5β1- or αvβ3-integrin to spread on fibronectin (FN) adsorbed to plastic. ECs in coculture lacked focal adhesions, but EC α5β1-integrin bound to fibrillar FN on the SMC surface, promoting rapid fibrillar adhesion formation. As assessed by both Western blot analysis and quantitative real-time RT-PCR, coculture suppressed the expression of focal adhesion proteins and mRNA, whereas tensin protein and mRNA expression were elevated. When attached to polyacrylamide gels with similar elastic moduli as SMCs, focal adhesion formation and the rate of cell spreading increased relative to ECs in coculture. Thus, the elastic properties are only one factor contributing to EC spreading and focal adhesion formation in coculture. The results suggest that the softness of the SMCs and the fibrillar organization of FN inhibit focal adhesions and reduce cell spreading while promoting fibrillar adhesion formation. These changes in the type of adhesions may alter EC signaling pathways in tissue-engineered blood vessels.

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ONE-DIMENSIONAL MODELING AND SIMULATIONS OF MIGRATION OF CULTURED FIBROBLASTS

Journal of Mechanics in Medicine and Biology ◽

10.1142/s0219519414500274 ◽

2014 ◽

Vol 14 (02) ◽

pp. 1450027

Author(s):

PEI-JUNG WU ◽

CHOU-CHING K. LIN ◽

MING-SHAUNG JU

Keyword(s):

Type I Collagen ◽

Model Simulation ◽

Adhesion Formation ◽

Fiber Formation ◽

Type I ◽

Cultured Fibroblasts ◽

One Dimensional ◽

3T3 Fibroblasts ◽

Kinematic Analyses ◽

Cell Front

Cell migration is crucial for many physiological functions such as wound healing, immuno-response and carcinogenesis. In this study an one-dimensional model of migration of fibroblasts was developed by modeling and integrating five subcellular processes, namely, actin protrusion, focal adhesion formation, stress fiber formation, polarization and retraction. The direction of migration was determined by polarization, which was related to direction of the stiffness gradient of the substrate. By controlling intensity of ultraviolet exposure on type-I collagen, a substrate with a stiffness gradient could be fabricated. Kinematic analyses of positions of the cell front, the nucleus and the cell rear, were utilized as inputs to the model. Simulation results of five live NIH 3T3 fibroblasts showed that the model was capable of simulating fast moving, slow moving and back-and-forth moving of the cells on the substrate.

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Transmembrane CEACAM1 affects integrin-dependent signaling and regulates extracellular matrix protein–specific morphology and migration of endothelial cells

Blood ◽

10.1182/blood-2004-09-3618 ◽

2005 ◽

Vol 105 (10) ◽

pp. 3925-3934 ◽

Cited By ~ 58

Author(s):

Mario M. Müller ◽

Bernhard B. Singer ◽

Esther Klaile ◽

Björn Öbrink ◽

Lothar Lucka

Keyword(s):

Endothelial Cells ◽

Cell Adhesion ◽

Focal Adhesion ◽

Matrix Protein ◽

Network Formation ◽

Adhesion Formation ◽

Fiber Formation ◽

Extracellular Matrix Protein ◽

And Migration ◽

Laminin 1

AbstractCarcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1/CD66a), expressed on leukocytes, epithelia, and endothelia mediates homophilic cell adhesion. It plays an important role in cell morphogenesis and, recently, soluble CEACAM1 isoforms have been implicated in angiogenesis. In the present study, we investigated the function of long transmembrane isoform of CEACAM1 (CEACAM1-L) in cultured rat brain endothelial cells. We observed that expression of CEACAM1-L promotes network formation on basement membrane Matrigel and increased cell motility after monolayer injury. During cell-matrix adhesion, CEACAM1-L translocated into the Triton X-100–insoluble cytoskeletal fraction and affected cell spreading and cell morphology on Matrigel and laminin-1 but not on fibronectin. On laminin-1, CEACAM1-L–expressing cells developed protrusions with lamellipodia, showed less stress fiber formation, reduced focal adhesion kinase (FAK) tyrosine phosphorylation, and decreased focal adhesion formation leading to high motility. CEACAM1-L–mediated morphologic alterations were sensitive to RhoA activation via lysophosphatidic acid (LPA) treatment and dependent on Rac1 activation. Furthermore, we demonstrate a matrix protein–dependent association of CEACAM1-L with talin, an important regulator of integrin function. Taken together, our results suggest that transmembrane CEACAM1-L expressed on endothelial cells is implicated in the activation phase of angiogenesis by affecting the cytoskeleton architecture and integrin-mediated signaling.

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