Regulation of Adhesion Strength by Focal Adhesion Position and Cell Shape

2012 ◽  
Author(s):  
Nathan D. Gallant ◽  
Kranthi Kumar Elineni
Keyword(s):  
Cell Adhesion ◽  
Adhesion Strength ◽  
Cell Shape ◽  
Microcontact Printing ◽  
Control Cell ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Spreading Area ◽  
Hydrodynamic Shear ◽  
Cell Spreading Area

Cell adhesion to extracellular matrices is critical to numerous cellular functions and is primarily mediated by integrin receptors. Binding and aggregation of integrins leads to the formation of focal adhesions (FA) which connect the cytoskeleton to the extracellular matrix in order to reinforce adhesion and transmit signals [1]. Preliminary observations indicated preferential recruitment of FAs to the periphery of the cell spreading area on both uniformly coated and micropatterned fibronectin surfaces (Fig. 1). The current study investigates the biophysical regulation of cell adhesion strength based on the size and position of FA with the central hypothesis that peripheral FAs stabilize adhesion strength. The hypothesis was tested by delineating the cell spreading area from the total cell adhesive area by employing microcontact printing to pattern substrates with a series of circular and annular adhesive islands which control cell shape (Fig. 2). A well characterized hydrodynamic shear assay known as the spinning disk device was used to quantify the adhesion strength of cells adhered to the micropatterns [2].

scholarly journals Focal Adhesions are Composed of Filamentous Subunits Whose Length and Dynamics Depend on the Cell Spreading Area

2014 ◽  
Vol 106 (2) ◽  
pp. 168a
Author(s):  
Shiqiong Hu ◽  
Yee-Han Tee ◽  
Alexandre Kabla ◽  
Alexander Bershadsky ◽  
Pascal Hersen
Keyword(s):  
Focal Adhesions ◽  
Cell Spreading ◽  
Spreading Area ◽  
Cell Spreading Area

scholarly journals Cell spreading area regulates clathrin-coated pit dynamics on micropatterned substrate

2015 ◽  
Vol 7 (9) ◽  
pp. 1033-1043 ◽  
Author(s):  
Xinyu Tan ◽  
Johanna Heureaux ◽  
Allen P. Liu
Keyword(s):  
Fluorescence Microscopy ◽  
Total Internal Reflection ◽  
Microcontact Printing ◽  
Cell Spreading ◽  
Internal Reflection ◽  
Total Internal Reflection Fluorescence ◽  
Spreading Area ◽  
Cell Spreading Area

Clathrin-coated pit dynamics changes with different cell spreading area revealed by microcontact printing and total internal reflection fluorescence microscopy.

scholarly journals p130Cas-associated Protein (p140Cap) as a New Tyrosine-phosphorylated Protein Involved in Cell Spreading

2004 ◽  
Vol 15 (2) ◽  
pp. 787-800 ◽  
Author(s):  
Paola Di Stefano ◽  
Sara Cabodi ◽  
Elisabetta Boeri Erba ◽  
Valentina Margaria ◽  
Elena Bergatto ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Growth Factor ◽  
Tyrosine Phosphorylation ◽  
Egf Receptor ◽  
Control Cell ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Growth Factor Signaling ◽  
Cell Extracts ◽  
Mouse Tissues

Integrin-mediated cell adhesion stimulates a cascade of signaling pathways that control cell proliferation, migration, and survival, mostly through tyrosine phosphorylation of signaling molecules. p130Cas, originally identified as a major substrate of v-Src, is a scaffold molecule that interacts with several proteins and mediates multiple cellular events after cell adhesion and mitogen treatment. Here, we describe a novel p130Cas-associated protein named p140Cap (Cas-associated protein) as a new tyrosine phosphorylated molecule involved in integrin- and epidermal growth factor (EGF)-dependent signaling. By affinity chromatography of human ECV304 cell extracts on a MBP-p130Cas column followed by mass spectrometry matrix-assisted laser desorption ionization/time of flight analysis, we identified p140Cap as a protein migrating at 140 kDa. We detected its expression in human, mouse, and rat cells and in different mouse tissues. Endogenous and transfected p140Cap proteins coimmunoprecipitate with p130Cas in ECV304 and in human embryonic kidney 293 cells and associate with p130Cas through their carboxy-terminal region. By immunofluorescence analysis, we demonstrated that in ECV304 cells plated on fibronectin, the endogenous p140Cap colocalizes with p130Cas in the perinuclear region as well as in lamellipodia. In addition p140Cap codistributes with cortical actin and actin stress fibers but not with focal adhesions. We also show that p140Cap is tyrosine phosphorylated within 15 min of cell adhesion to integrin ligands. p140Cap tyrosine phosphorylation is also induced in response to EGF through an EGF receptor dependent-mechanism. Interestingly expression of p140Cap in NIH3T3 and in ECV304 cells delays the onset of cell spreading in the early phases of cell adhesion to fibronectin. Therefore, p140Cap is a novel protein associated with p130Cas and actin cytoskeletal structures. Its tyrosine phosphorylation by integrin-mediated adhesion and EGF stimulation and its involvement in cell spreading on matrix proteins suggest that p140Cap plays a role in controlling actin cytoskeleton organization in response to adhesive and growth factor signaling.

scholarly journals Desmoglein-2 harnesses a PDZ-GEF2/Rap1 signaling axis to control cell spreading and focal adhesions independent of cell–cell adhesion

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
W. Tucker Shelton ◽  
S. Madison Thomas ◽  
Hunter R. Alexander ◽  
C. Evan Thomes ◽  
Daniel E. Conway ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Control Cell ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Cell Junctions ◽  
A431 Cells ◽  
Signaling Crosstalk ◽  
Cell Matrix ◽  
And Migration ◽  
Cell Cell

AbstractDesmosomes have a central role in mediating extracellular adhesion between cells, but they also coordinate other biological processes such as proliferation, differentiation, apoptosis and migration. In particular, several lines of evidence have implicated desmosomal proteins in regulating the actin cytoskeleton and attachment to the extracellular matrix, indicating signaling crosstalk between cell–cell junctions and cell–matrix adhesions. In our study, we found that cells lacking the desmosomal cadherin Desmoglein-2 (Dsg2) displayed a significant increase in spreading area on both fibronectin and collagen, compared to control A431 cells. Intriguingly, this effect was observed in single spreading cells, indicating that Dsg2 can exert its effects on cell spreading independent of cell–cell adhesion. We hypothesized that Dsg2 may mediate cell–matrix adhesion via control of Rap1 GTPase, which is well known as a central regulator of cell spreading dynamics. We show that Rap1 activity is elevated in Dsg2 knockout cells, and that Dsg2 harnesses Rap1 and downstream TGFβ signaling to influence both cell spreading and focal adhesion protein phosphorylation. Further analysis implicated the Rap GEF PDZ-GEF2 in mediating Dsg2-dependent cell spreading. These data have identified a novel role for Dsg2 in controlling cell spreading, providing insight into the mechanisms via which cadherins exert non-canonical junction-independent effects.

Cell spreading and the regulation of ornithine decarboxylase

1995 ◽  
Vol 108 (12) ◽  
pp. 3787-3794
Author(s):  
R.F. Morrison ◽  
E.R. Seidel
Keyword(s):  
Tissue Culture ◽  
Cell Adhesion ◽  
Ornithine Decarboxylase ◽  
Cell Spreading ◽  
Decarboxylase Activity ◽  
Spreading Area ◽  
Ornithine Decarboxylase Activity ◽  
The Mean ◽  
Low Cell Density ◽  
Dose Dependent

The aim of this study was to investigate the effect of cell spreading on the induction of ornithine decarboxylase and the rate of putrescine uptake in anchorage-dependent and anchorage-independent cells. Plating non-transformed IEC-6 epithelial cells at high versus low cell density restricted cell spreading from 900 microns 2 to approximately 140 microns 2, blunted the transient induction of ornithine decarboxylase activity from 202 to 32 pmol 14CO2/mg protein per hour and reduced the rate of [14C] putrescine uptake from 46 to 23 pmol/10(5) cells per hour. The mean spreading area of the cell population was controlled by coating tissue culture dishes with the nonadhesive polymer, polyHEMA. Ornithine decarboxylase activity and putrescine uptake correlated with cell spreading with minimal spreading (263 microns 2) corresponding to an 83% decrease in ornithine decarboxylase activity and 51% decrease in the rate of putrescine uptake. Adding the RGD peptide, Gly-Arg-Gly-Glu-Ser-Pro to the medium of sparsely plated cells resulted in rapid reductions in cell spreading concomitant with dose-dependent decreases in ornithine decarboxylase activity and putrescine uptake. Finally, minimizing cell spreading by depriving cells of substratum contact completely abolished serum-induced increases in ornithine decarboxylase and reduced the rate of putrescine uptake by 47%. In contrast to IEC-6 cells, ornithine decarboxylase of neoplastic HTC-116 cells was constitutively expressed with basal and stimulated activity (193 and 982 pmol 14CO2/mg protein per hour, respectively) completely independent of cell adhesion. Putrescine uptake, however, was abolished in the absence of cell adhesion. These data suggest that the induction of ornithine decarboxylase activity and the rate of putrescine uptake correlate with spreading of anchorage-dependent IEC-6 cells and that ornithine decarboxylase activity but not putrescine uptake, appears to be independent of spreading of neoplastic HTC-116 cells.

scholarly journals Topographic cell instructive patterns to control cell adhesion, polarization and migration

2014 ◽  
Vol 11 (100) ◽  
pp. 20140687 ◽  
Author(s):  
Maurizio Ventre ◽  
Carlo Fortunato Natale ◽  
Carmela Rianna ◽  
Paolo Antonio Netti
Keyword(s):  
Cell Adhesion ◽  
Control Cell ◽  
Chemical Properties ◽  
Focal Adhesions ◽  
Adhesive Properties ◽  
Cellular Processes ◽  
Topographic Features ◽  
And Migration ◽  
Specific Length ◽  
And Chemical Properties

Topographic patterns are known to affect cellular processes such as adhesion, migration and differentiation. However, the optimal way to deliver topographic signals to provide cells with precise instructions has not been defined yet. In this work, we hypothesize that topographic patterns may be able to control the sensing and adhesion machinery of cells when their interval features are tuned on the characteristic lengths of filopodial probing and focal adhesions (FAs). Features separated by distance beyond the length of filopodia cannot be readily perceived; therefore, the formation of new adhesions is discouraged. If, however, topographic features are separated by a distance within the reach of filopodia extension, cells can establish contact between adjacent topographic islands. In the latter case, cell adhesion and polarization rely upon the growth of FAs occurring on a specific length scale that depends on the chemical properties of the surface. Topographic patterns and chemical properties may interfere with the growth of FAs, thus making adhesions unstable. To test this hypothesis, we fabricated different micropatterned surfaces displaying feature dimensions and adhesive properties able to interfere with the filopodial sensing and the adhesion maturation, selectively. Our data demonstrate that it is possible to exert a potent control on cell adhesion, elongation and migration by tuning topographic features’ dimensions and surface chemistry.

Tuning epithelial cell-cell adhesion and collective dynamics with functional DNA-E-cadherin hybrid linkers

2021 ◽  
Author(s):  
Andreas Schoenit ◽  
Cristina Lo Giudice ◽  
Nina Hahnen ◽  
Dirk Ollech ◽  
Kevin Jahnke ◽  
...  
Keyword(s):  
Cell Adhesion ◽  
Epithelial Cells ◽  
Adhesion Strength ◽  
Control Cell ◽  
Dna Nanotechnology ◽  
Cell Dynamics ◽  
Binding Strength ◽  
Dna Sequence Design ◽  
E Cadherin ◽  
Cell Cell

The binding strength between epithelial cells is crucial for tissue integrity, signal transduction and collective cell dynamics. However, there is no experimental approach to precisely modulate cell-cell adhesion strength at the cellular and molecular level. Here, we establish DNA nanotechnology as tool to control cell-cell adhesion of epithelial cells. We designed a DNA-E-cadherin hybrid system consisting of complementary DNA strands covalently bound to a truncated E-cadherin with a modified extracellular domain. DNA sequence design allows to tune the DNA-E-cadherin hybrid molecular binding strength, while retaining its cytosolic interactions and downstream signaling capabilities. The DNA-E-cadherin hybrid facilitates strong and reversible cell-cell adhesion in E-cadherin deficient cells by forming mechanotransducive adherens junctions. We assess the direct influence of cell-cell adhesion strength on intracellular signaling and collective cell dynamics. This highlights the scope of DNA nanotechnology as a precision technology to study and engineer cell collectives.

scholarly journals Keeping the Vimentin Network under Control: Cell–Matrix Adhesion–associated Plectin 1f Affects Cell Shape and Polarity of Fibroblasts

2010 ◽  
Vol 21 (19) ◽  
pp. 3362-3375 ◽  
Author(s):  
Gerald Burgstaller ◽  
Martin Gregor ◽  
Lilli Winter ◽  
Gerhard Wiche
Keyword(s):  
Cell Shape ◽  
Network Formation ◽  
De Novo ◽  
Control Cell ◽  
Focal Adhesions ◽  
Time Lapse ◽  
Cell Polarization ◽  
Dependent Manner ◽  
Cell Matrix ◽  
Contractile Stress

Focal adhesions (FAs) located at the ends of actin/myosin-containing contractile stress fibers form tight connections between fibroblasts and their underlying extracellular matrix. We show here that mature FAs and their derivative fibronectin fibril-aligned fibrillar adhesions (FbAs) serve as docking sites for vimentin intermediate filaments (IFs) in a plectin isoform 1f (P1f)-dependent manner. Time-lapse video microscopy revealed that FA-associated P1f captures mobile vimentin filament precursors, which then serve as seeds for de novo IF network formation via end-to-end fusion with other mobile precursors. As a consequence of IF association, the turnover of FAs is reduced. P1f-mediated IF network formation at FbAs creates a resilient cage-like core structure that encases and positions the nucleus while being stably connected to the exterior of the cell. We show that the formation of this structure affects cell shape with consequences for cell polarization.

scholarly journals Influence of Cell Spreading Area on the Osteogenic Commitment and Phenotype Maintenance of Mesenchymal Stem Cells

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yingjun Yang ◽  
Xinlong Wang ◽  
Yongtao Wang ◽  
Xiaohong Hu ◽  
Naoki Kawazoe ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Spreading ◽  
Spreading Area ◽  
Cell Spreading Area

scholarly journals Impairment of cell adhesion and migration by inhibition of protein disulphide isomerases in three breast cancer cell lines

2020 ◽  
Vol 40 (10) ◽  
Author(s):  
Henry S. Young ◽  
Lucy M. McGowan ◽  
Katy A. Jepson ◽  
Josephine C. Adams
Keyword(s):  
Breast Cancer ◽  
Cell Adhesion ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
Focal Adhesions ◽  
Cell Spreading ◽  
Secreted Proteins ◽  
Breast Cancer Cell Lines ◽  
Scratch Wound ◽  
And Migration

Abstract Protein disulphide isomerase A3 (PDIA3) is an endoplasmic reticulum (ER)-resident disulphide isomerase and oxidoreductase with known substrates that include some extracellular matrix (ECM) proteins. PDIA3 is up-regulated in invasive breast cancers and correlates in a mouse orthotopic xenograft model with breast cancer metastasis to bone. However, the underlying cellular mechanisms remain unclear. Here we investigated the function of protein disulphide isomerases in attachment, spreading and migration of three human breast cancer lines representative of luminal (MCF-7) or basal (MDA-MB-231 and HCC1937) tumour phenotypes. Pharmacological inhibition by 16F16 decreased initial cell spreading more effectively than inhibition by PACMA-31. Cells displayed diminished cortical F-actin projections, stress fibres and focal adhesions. Cell migration was reduced in a quantified ‘scratch wound’ assay. To examine whether these effects might result from alterations to secreted proteins in the absence of functional PDIA3, adhesion and migration were quantified in the above cells exposed to media conditioned by wildtype (WT) or Pdia3−/− mouse embryonic fibroblasts (MEFs). The conditioned medium (CM) of Pdia3−/− MEFs was less effective in promoting cell spreading and F-actin organisation or supporting ‘scratch wound’ closure. Similarly, ECM prepared from HCC1937 cells after 16F16 inhibition was less effective than control ECM to support spreading of untreated HCC1937 cells. Overall, these results advance the concept that protein disulphide isomerases including PDIA3 drive the production of secreted proteins that promote a microenvironment favourable to breast cancer cell adhesion and motility, characteristics that are integral to tumour invasion and metastasis. Inhibition of PDIA3 or related isomerases may have potential for anti-metastatic therapies.

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