Limits...
Integrins control motile strategy through a Rho-cofilin pathway.

Danen EH, van Rheenen J, Franken W, Huveneers S, Sonneveld P, Jalink K, Sonnenberg A - J. Cell Biol. (2005)

Bottom Line: During wound healing, angiogenesis, and tumor invasion, cells often change their expression profiles of fibronectin-binding integrins.The activity of the small GTPase RhoA is particularly high in cells adhering by alpha5beta1, and inhibition of Rho signaling causes a switch from a beta1- to a beta3-associated mode of migration, whereas increased Rho activity has the opposite effect.Thus, alterations in integrin expression profiles allow cells to modulate several critical aspects of the motile machinery through Rho GTPases.

View Article: PubMed Central - PubMed

Affiliation: Division of Cell Biology, Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands. e.danen@nki.nl

ABSTRACT
During wound healing, angiogenesis, and tumor invasion, cells often change their expression profiles of fibronectin-binding integrins. Here, we show that beta1 integrins promote random migration, whereas beta3 integrins promote persistent migration in the same epithelial cell background. Adhesion to fibronectin by alpha(v)beta3 supports extensive actin cytoskeletal reorganization through the actin-severing protein cofilin, resulting in a single broad lamellipod with static cell-matrix adhesions at the leading edge. Adhesion by alpha5beta1 instead leads to the phosphorylation/inactivation of cofilin, and these cells fail to polarize their cytoskeleton but extend thin protrusions containing highly dynamic cell-matrix adhesions in multiple directions. The activity of the small GTPase RhoA is particularly high in cells adhering by alpha5beta1, and inhibition of Rho signaling causes a switch from a beta1- to a beta3-associated mode of migration, whereas increased Rho activity has the opposite effect. Thus, alterations in integrin expression profiles allow cells to modulate several critical aspects of the motile machinery through Rho GTPases.

Show MeSH

Related in: MedlinePlus

β1 and β3 integrins differentially affect motile behavior. (A and B) Migration tracks of GEβ1 (A) or GEβ3 cells (B) seeded sparsely on FN-coated coverslips and followed for 16 h. Shown are 20 cells obtained from three independent experiments. (C) Analysis of persistence (ratio of the direct distance from start point to end point divided by the total track distance) and speed of migrating GEβ1 and GEβ3 cells in sparse cultures. (D) Analysis of MTOC polarization in wounding assays with GEβ1 (filled bars) and GEβ3 cells (open bars) at the indicated time points after wounding of confluent monolayers on FN-coated coverslips. Mean ± SD of ∼100 cells analyzed in three independent assays is shown. See supplemental data for the accompanying videos and immunofluorescence images (available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2171933&req=5

fig1: β1 and β3 integrins differentially affect motile behavior. (A and B) Migration tracks of GEβ1 (A) or GEβ3 cells (B) seeded sparsely on FN-coated coverslips and followed for 16 h. Shown are 20 cells obtained from three independent experiments. (C) Analysis of persistence (ratio of the direct distance from start point to end point divided by the total track distance) and speed of migrating GEβ1 and GEβ3 cells in sparse cultures. (D) Analysis of MTOC polarization in wounding assays with GEβ1 (filled bars) and GEβ3 cells (open bars) at the indicated time points after wounding of confluent monolayers on FN-coated coverslips. Mean ± SD of ∼100 cells analyzed in three independent assays is shown. See supplemental data for the accompanying videos and immunofluorescence images (available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1).

Mentions: Notably, the motile behavior in wounding assays differed dramatically for cells adhering by α5β1 or αvβ3: GEβ1 cells moved randomly as single cells extending protrusions in multiple directions, whereas GEβ3 cells moved as a sheet of cells that maintained directionality (Video 1). To rule out effects due to differences in cell–cell adhesion, the migration of sparsely seeded cells was analyzed. The velocity under these conditions was ∼25 μm/h for both cell types. However, similar to the results obtained in wounding assays, GEβ1 cells moved randomly whereas GEβ3 cells moved in a much more persistent fashion (Fig. 1 A–C; Video 2, available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1). To examine if the lack of directionality of GEβ1 cells in wounding assays was due to a defect in polarization, we analyzed if these cells responded to wounding by polarizing their microtubule-organizing center (MTOC) (Gotlieb et al., 1981). GEβ1 and GEβ3 cells at the wound edge polarized their MTOC with similar efficiency within 3 h after wounding (Fig. 1 D; Fig. S1, available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1), indicating that GEβ1 cells could “sense” the wound and orient their MTOC accordingly. However, although GEβ3 cells maintained their polarized phenotype as they migrated into the wound area, MTOC polarity in the direction of the wound was lost in GEβ1 cells during this process (Fig. 1 D, 9 h).


Integrins control motile strategy through a Rho-cofilin pathway.

Danen EH, van Rheenen J, Franken W, Huveneers S, Sonneveld P, Jalink K, Sonnenberg A - J. Cell Biol. (2005)

β1 and β3 integrins differentially affect motile behavior. (A and B) Migration tracks of GEβ1 (A) or GEβ3 cells (B) seeded sparsely on FN-coated coverslips and followed for 16 h. Shown are 20 cells obtained from three independent experiments. (C) Analysis of persistence (ratio of the direct distance from start point to end point divided by the total track distance) and speed of migrating GEβ1 and GEβ3 cells in sparse cultures. (D) Analysis of MTOC polarization in wounding assays with GEβ1 (filled bars) and GEβ3 cells (open bars) at the indicated time points after wounding of confluent monolayers on FN-coated coverslips. Mean ± SD of ∼100 cells analyzed in three independent assays is shown. See supplemental data for the accompanying videos and immunofluorescence images (available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2171933&req=5

fig1: β1 and β3 integrins differentially affect motile behavior. (A and B) Migration tracks of GEβ1 (A) or GEβ3 cells (B) seeded sparsely on FN-coated coverslips and followed for 16 h. Shown are 20 cells obtained from three independent experiments. (C) Analysis of persistence (ratio of the direct distance from start point to end point divided by the total track distance) and speed of migrating GEβ1 and GEβ3 cells in sparse cultures. (D) Analysis of MTOC polarization in wounding assays with GEβ1 (filled bars) and GEβ3 cells (open bars) at the indicated time points after wounding of confluent monolayers on FN-coated coverslips. Mean ± SD of ∼100 cells analyzed in three independent assays is shown. See supplemental data for the accompanying videos and immunofluorescence images (available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1).
Mentions: Notably, the motile behavior in wounding assays differed dramatically for cells adhering by α5β1 or αvβ3: GEβ1 cells moved randomly as single cells extending protrusions in multiple directions, whereas GEβ3 cells moved as a sheet of cells that maintained directionality (Video 1). To rule out effects due to differences in cell–cell adhesion, the migration of sparsely seeded cells was analyzed. The velocity under these conditions was ∼25 μm/h for both cell types. However, similar to the results obtained in wounding assays, GEβ1 cells moved randomly whereas GEβ3 cells moved in a much more persistent fashion (Fig. 1 A–C; Video 2, available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1). To examine if the lack of directionality of GEβ1 cells in wounding assays was due to a defect in polarization, we analyzed if these cells responded to wounding by polarizing their microtubule-organizing center (MTOC) (Gotlieb et al., 1981). GEβ1 and GEβ3 cells at the wound edge polarized their MTOC with similar efficiency within 3 h after wounding (Fig. 1 D; Fig. S1, available at http://www.jcb.org/cgi/content/full/jcb.200412081/DC1), indicating that GEβ1 cells could “sense” the wound and orient their MTOC accordingly. However, although GEβ3 cells maintained their polarized phenotype as they migrated into the wound area, MTOC polarity in the direction of the wound was lost in GEβ1 cells during this process (Fig. 1 D, 9 h).

Bottom Line: During wound healing, angiogenesis, and tumor invasion, cells often change their expression profiles of fibronectin-binding integrins.The activity of the small GTPase RhoA is particularly high in cells adhering by alpha5beta1, and inhibition of Rho signaling causes a switch from a beta1- to a beta3-associated mode of migration, whereas increased Rho activity has the opposite effect.Thus, alterations in integrin expression profiles allow cells to modulate several critical aspects of the motile machinery through Rho GTPases.

View Article: PubMed Central - PubMed

Affiliation: Division of Cell Biology, Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands. e.danen@nki.nl

ABSTRACT
During wound healing, angiogenesis, and tumor invasion, cells often change their expression profiles of fibronectin-binding integrins. Here, we show that beta1 integrins promote random migration, whereas beta3 integrins promote persistent migration in the same epithelial cell background. Adhesion to fibronectin by alpha(v)beta3 supports extensive actin cytoskeletal reorganization through the actin-severing protein cofilin, resulting in a single broad lamellipod with static cell-matrix adhesions at the leading edge. Adhesion by alpha5beta1 instead leads to the phosphorylation/inactivation of cofilin, and these cells fail to polarize their cytoskeleton but extend thin protrusions containing highly dynamic cell-matrix adhesions in multiple directions. The activity of the small GTPase RhoA is particularly high in cells adhering by alpha5beta1, and inhibition of Rho signaling causes a switch from a beta1- to a beta3-associated mode of migration, whereas increased Rho activity has the opposite effect. Thus, alterations in integrin expression profiles allow cells to modulate several critical aspects of the motile machinery through Rho GTPases.

Show MeSH
Related in: MedlinePlus