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Integrin-dependent actomyosin contraction regulates epithelial cell scattering.

de Rooij J, Kerstens A, Danuser G, Schwartz MA, Waterman-Storer CM - J. Cell Biol. (2005)

Bottom Line: Scattering is enhanced on collagen and fibronectin, as compared with laminin1, suggesting possible cross talk between integrins and cell-cell junctions.Rigid substrates that produce high traction forces promoted scattering, in comparison to more compliant substrates.We conclude that integrin-dependent actomyosin traction force mediates the disruption of cell-cell adhesion during epithelial cell scattering.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

ABSTRACT
The scattering of Madin-Darby canine kidney cells in vitro mimics key aspects of epithelial-mesenchymal transitions during development, carcinoma cell invasion, and metastasis. Scattering is induced by hepatocyte growth factor (HGF) and is thought to involve disruption of cadherin-dependent cell-cell junctions. Scattering is enhanced on collagen and fibronectin, as compared with laminin1, suggesting possible cross talk between integrins and cell-cell junctions. We show that HGF does not trigger any detectable decrease in E-cadherin function, but increases integrin-mediated adhesion. Time-lapse imaging suggests that tension on cell-cell junctions may disrupt cell-cell adhesion. Varying the density and type of extracellular matrix proteins shows that scattering correlates with stronger integrin adhesion and increased phosphorylation of the myosin regulatory light chain. To directly test the role of integrin-dependent traction forces, substrate compliance was varied. Rigid substrates that produce high traction forces promoted scattering, in comparison to more compliant substrates. We conclude that integrin-dependent actomyosin traction force mediates the disruption of cell-cell adhesion during epithelial cell scattering.

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HGF does not inhibit the ability of E-cadherin to form homotypic interactions. (A) Lack of effect of HGF on E-cadherin in cell–cell junctions. MDCK cells plated at high or low density on Cn-coated coverslips were treated with HGF for 24 h. Cells were fixed and E-cadherin was immunolocalized. (B) Lack of effect of HGF on GFP-E-cadherin distribution on E-cadherin–coated coverslips. GFP-E-cadherin–expressing MDCK cells were plated on Ecad-comp in the absence of HGF for 3 h or in the presence of HGF for 24 h, fixed, and stained for F-actin. (C) HGF increases adhesion to Ecad-comp. MDCK cells were plated on the indicated amounts of Ecad-comp in the absence or presence of HGF or inhibitors (DECMA-1 or HAV-peptide) and treated and quantified as described in Materials and methods. Data are means ± SD; n = 3.
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fig1: HGF does not inhibit the ability of E-cadherin to form homotypic interactions. (A) Lack of effect of HGF on E-cadherin in cell–cell junctions. MDCK cells plated at high or low density on Cn-coated coverslips were treated with HGF for 24 h. Cells were fixed and E-cadherin was immunolocalized. (B) Lack of effect of HGF on GFP-E-cadherin distribution on E-cadherin–coated coverslips. GFP-E-cadherin–expressing MDCK cells were plated on Ecad-comp in the absence of HGF for 3 h or in the presence of HGF for 24 h, fixed, and stained for F-actin. (C) HGF increases adhesion to Ecad-comp. MDCK cells were plated on the indicated amounts of Ecad-comp in the absence or presence of HGF or inhibitors (DECMA-1 or HAV-peptide) and treated and quantified as described in Materials and methods. Data are means ± SD; n = 3.

Mentions: It has generally been supposed that HGF disrupts E-cadherin function to induce the loss of cell–cell adhesion and increase cell migration. To investigate the effect of HGF on cell–cell adhesion in the absence of cell migration, cells were plated either densely enough to form a continuous monolayer or sparsely enough to allow the formation of isolated islands of cells. In both cases, coverslips were coated with 3 μg/ml Cn, which strongly promotes scattering, and cells were plated with HGF 24 h before stimulation. 20 h after HGF stimulation, cells were fixed and stained for E-cadherin and F-actin. In densely plated cells, E-cadherin clearly remained in cell–cell junctions at an intensity similar to that in unstimulated cells (Fig. 1 A). When sparsely plated, most cells, as expected, broke contacts with their neighboring cells and moved out of the epithelial islands so that the area of cell–cell contact decreased. However, the remaining cell–cell junctions clearly retained E-cadherin also at an intensity similar to that in unstimulated cells. These results suggest that the disruption of E-cadherin–based cell–cell junctions by HGF is dependent on the ability of cells to migrate and move apart from one another and does not involve direct reduction in E-cadherin function.


Integrin-dependent actomyosin contraction regulates epithelial cell scattering.

de Rooij J, Kerstens A, Danuser G, Schwartz MA, Waterman-Storer CM - J. Cell Biol. (2005)

HGF does not inhibit the ability of E-cadherin to form homotypic interactions. (A) Lack of effect of HGF on E-cadherin in cell–cell junctions. MDCK cells plated at high or low density on Cn-coated coverslips were treated with HGF for 24 h. Cells were fixed and E-cadherin was immunolocalized. (B) Lack of effect of HGF on GFP-E-cadherin distribution on E-cadherin–coated coverslips. GFP-E-cadherin–expressing MDCK cells were plated on Ecad-comp in the absence of HGF for 3 h or in the presence of HGF for 24 h, fixed, and stained for F-actin. (C) HGF increases adhesion to Ecad-comp. MDCK cells were plated on the indicated amounts of Ecad-comp in the absence or presence of HGF or inhibitors (DECMA-1 or HAV-peptide) and treated and quantified as described in Materials and methods. Data are means ± SD; n = 3.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2171213&req=5

fig1: HGF does not inhibit the ability of E-cadherin to form homotypic interactions. (A) Lack of effect of HGF on E-cadherin in cell–cell junctions. MDCK cells plated at high or low density on Cn-coated coverslips were treated with HGF for 24 h. Cells were fixed and E-cadherin was immunolocalized. (B) Lack of effect of HGF on GFP-E-cadherin distribution on E-cadherin–coated coverslips. GFP-E-cadherin–expressing MDCK cells were plated on Ecad-comp in the absence of HGF for 3 h or in the presence of HGF for 24 h, fixed, and stained for F-actin. (C) HGF increases adhesion to Ecad-comp. MDCK cells were plated on the indicated amounts of Ecad-comp in the absence or presence of HGF or inhibitors (DECMA-1 or HAV-peptide) and treated and quantified as described in Materials and methods. Data are means ± SD; n = 3.
Mentions: It has generally been supposed that HGF disrupts E-cadherin function to induce the loss of cell–cell adhesion and increase cell migration. To investigate the effect of HGF on cell–cell adhesion in the absence of cell migration, cells were plated either densely enough to form a continuous monolayer or sparsely enough to allow the formation of isolated islands of cells. In both cases, coverslips were coated with 3 μg/ml Cn, which strongly promotes scattering, and cells were plated with HGF 24 h before stimulation. 20 h after HGF stimulation, cells were fixed and stained for E-cadherin and F-actin. In densely plated cells, E-cadherin clearly remained in cell–cell junctions at an intensity similar to that in unstimulated cells (Fig. 1 A). When sparsely plated, most cells, as expected, broke contacts with their neighboring cells and moved out of the epithelial islands so that the area of cell–cell contact decreased. However, the remaining cell–cell junctions clearly retained E-cadherin also at an intensity similar to that in unstimulated cells. These results suggest that the disruption of E-cadherin–based cell–cell junctions by HGF is dependent on the ability of cells to migrate and move apart from one another and does not involve direct reduction in E-cadherin function.

Bottom Line: Scattering is enhanced on collagen and fibronectin, as compared with laminin1, suggesting possible cross talk between integrins and cell-cell junctions.Rigid substrates that produce high traction forces promoted scattering, in comparison to more compliant substrates.We conclude that integrin-dependent actomyosin traction force mediates the disruption of cell-cell adhesion during epithelial cell scattering.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

ABSTRACT
The scattering of Madin-Darby canine kidney cells in vitro mimics key aspects of epithelial-mesenchymal transitions during development, carcinoma cell invasion, and metastasis. Scattering is induced by hepatocyte growth factor (HGF) and is thought to involve disruption of cadherin-dependent cell-cell junctions. Scattering is enhanced on collagen and fibronectin, as compared with laminin1, suggesting possible cross talk between integrins and cell-cell junctions. We show that HGF does not trigger any detectable decrease in E-cadherin function, but increases integrin-mediated adhesion. Time-lapse imaging suggests that tension on cell-cell junctions may disrupt cell-cell adhesion. Varying the density and type of extracellular matrix proteins shows that scattering correlates with stronger integrin adhesion and increased phosphorylation of the myosin regulatory light chain. To directly test the role of integrin-dependent traction forces, substrate compliance was varied. Rigid substrates that produce high traction forces promoted scattering, in comparison to more compliant substrates. We conclude that integrin-dependent actomyosin traction force mediates the disruption of cell-cell adhesion during epithelial cell scattering.

Show MeSH
Related in: MedlinePlus