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One-dimensional topography underlies three-dimensional fibrillar cell migration.

Doyle AD, Wang FW, Matsumoto K, Yamada KM - J. Cell Biol. (2009)

Bottom Line: We use a novel micropatterning technique termed microphotopatterning (microPP) to identify functions for 1D fibrillar patterns in 3D cell migration.In striking contrast to 2D, cell migration in both 1D and 3D is rapid, uniaxial, independent of ECM ligand density, and dependent on myosin II contractility and microtubules (MTs). 1D and 3D migration are also characterized by an anterior MT bundle with a posterior centrosome.We propose that cells migrate rapidly through 3D fibrillar matrices by a 1D migratory mechanism not mimicked by 2D matrices.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA. adoyle@mail.nih.gov

ABSTRACT
Current concepts of cell migration were established in regular two-dimensional (2D) cell culture, but the roles of topography are poorly understood for cells migrating in an oriented 3D fibrillar extracellular matrix (ECM). We use a novel micropatterning technique termed microphotopatterning (microPP) to identify functions for 1D fibrillar patterns in 3D cell migration. In striking contrast to 2D, cell migration in both 1D and 3D is rapid, uniaxial, independent of ECM ligand density, and dependent on myosin II contractility and microtubules (MTs). 1D and 3D migration are also characterized by an anterior MT bundle with a posterior centrosome. We propose that cells migrate rapidly through 3D fibrillar matrices by a 1D migratory mechanism not mimicked by 2D matrices.

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Regulation of cell migration by fiber width and dimensionality. (A) Triplets (bracketed) of 1, 2.5, 5, 15, and 20 µm-wide lines used for cell migration tracking. (B) Biphasic effect of fiber width on fibrillar migration rate. (C) TIRF images of GFP-vinculin containing adhesions on a 2.5-µm line show two parallel sets of adhesions. The arrow indicates the direction of migration. (D) Fibroblasts migrating with a 3D/1D phenotype along multiple 1-µm lines. (E) 1D to 2D ECM transition induces spreading of the leading edge (arrow) and formation of many cell processes (arrowheads). The red box indicates ROI in time lapse below. (F) Rac siRNA knockdown in 2D induces a uniaxial phenotype lacking the single linear adhesion seen in 1D and does not increase migration rate. *, P < 0.05 versus all conditions except multilines; +, P < 0.05 versus 15–40 µm. Con, control. Error bars indicate SEM. Bars, 10 µm.
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fig4: Regulation of cell migration by fiber width and dimensionality. (A) Triplets (bracketed) of 1, 2.5, 5, 15, and 20 µm-wide lines used for cell migration tracking. (B) Biphasic effect of fiber width on fibrillar migration rate. (C) TIRF images of GFP-vinculin containing adhesions on a 2.5-µm line show two parallel sets of adhesions. The arrow indicates the direction of migration. (D) Fibroblasts migrating with a 3D/1D phenotype along multiple 1-µm lines. (E) 1D to 2D ECM transition induces spreading of the leading edge (arrow) and formation of many cell processes (arrowheads). The red box indicates ROI in time lapse below. (F) Rac siRNA knockdown in 2D induces a uniaxial phenotype lacking the single linear adhesion seen in 1D and does not increase migration rate. *, P < 0.05 versus all conditions except multilines; +, P < 0.05 versus 15–40 µm. Con, control. Error bars indicate SEM. Bars, 10 µm.

Mentions: A potential explanation for the persistent high velocities we observed with 1D fibrillar migration could be physical limitation of lateral cell spreading and lateral lamellae. This proposal is supported by analysis of cell adhesion area as measured by vinculin staining and total spread area, both of which were significantly reduced in 1D compared with 2D conditions (adhesion area, 54.6 ± 22.1 vs. 89.9 ± 37.4; total spread area, 370.5 ± 183 vs. 1,414 ± 630; P < 0.05). To test this hypothesis further, we generated fiberlike patterns with widths of 1–40 µm (Fig. 4 A). With increasing fibril width, cells generally continued to migrate in one direction but demonstrated increased cell spreading, loss of uniaxial morphology above 5-µm fibril widths, and decreased migration rate (Fig. 4 B). These data indicate that loss of the uniaxial phenotype in conjunction with increased lateral cell spreading on wider fibrils is detrimental to rapid cell migration.


One-dimensional topography underlies three-dimensional fibrillar cell migration.

Doyle AD, Wang FW, Matsumoto K, Yamada KM - J. Cell Biol. (2009)

Regulation of cell migration by fiber width and dimensionality. (A) Triplets (bracketed) of 1, 2.5, 5, 15, and 20 µm-wide lines used for cell migration tracking. (B) Biphasic effect of fiber width on fibrillar migration rate. (C) TIRF images of GFP-vinculin containing adhesions on a 2.5-µm line show two parallel sets of adhesions. The arrow indicates the direction of migration. (D) Fibroblasts migrating with a 3D/1D phenotype along multiple 1-µm lines. (E) 1D to 2D ECM transition induces spreading of the leading edge (arrow) and formation of many cell processes (arrowheads). The red box indicates ROI in time lapse below. (F) Rac siRNA knockdown in 2D induces a uniaxial phenotype lacking the single linear adhesion seen in 1D and does not increase migration rate. *, P < 0.05 versus all conditions except multilines; +, P < 0.05 versus 15–40 µm. Con, control. Error bars indicate SEM. Bars, 10 µm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2654121&req=5

fig4: Regulation of cell migration by fiber width and dimensionality. (A) Triplets (bracketed) of 1, 2.5, 5, 15, and 20 µm-wide lines used for cell migration tracking. (B) Biphasic effect of fiber width on fibrillar migration rate. (C) TIRF images of GFP-vinculin containing adhesions on a 2.5-µm line show two parallel sets of adhesions. The arrow indicates the direction of migration. (D) Fibroblasts migrating with a 3D/1D phenotype along multiple 1-µm lines. (E) 1D to 2D ECM transition induces spreading of the leading edge (arrow) and formation of many cell processes (arrowheads). The red box indicates ROI in time lapse below. (F) Rac siRNA knockdown in 2D induces a uniaxial phenotype lacking the single linear adhesion seen in 1D and does not increase migration rate. *, P < 0.05 versus all conditions except multilines; +, P < 0.05 versus 15–40 µm. Con, control. Error bars indicate SEM. Bars, 10 µm.
Mentions: A potential explanation for the persistent high velocities we observed with 1D fibrillar migration could be physical limitation of lateral cell spreading and lateral lamellae. This proposal is supported by analysis of cell adhesion area as measured by vinculin staining and total spread area, both of which were significantly reduced in 1D compared with 2D conditions (adhesion area, 54.6 ± 22.1 vs. 89.9 ± 37.4; total spread area, 370.5 ± 183 vs. 1,414 ± 630; P < 0.05). To test this hypothesis further, we generated fiberlike patterns with widths of 1–40 µm (Fig. 4 A). With increasing fibril width, cells generally continued to migrate in one direction but demonstrated increased cell spreading, loss of uniaxial morphology above 5-µm fibril widths, and decreased migration rate (Fig. 4 B). These data indicate that loss of the uniaxial phenotype in conjunction with increased lateral cell spreading on wider fibrils is detrimental to rapid cell migration.

Bottom Line: We use a novel micropatterning technique termed microphotopatterning (microPP) to identify functions for 1D fibrillar patterns in 3D cell migration.In striking contrast to 2D, cell migration in both 1D and 3D is rapid, uniaxial, independent of ECM ligand density, and dependent on myosin II contractility and microtubules (MTs). 1D and 3D migration are also characterized by an anterior MT bundle with a posterior centrosome.We propose that cells migrate rapidly through 3D fibrillar matrices by a 1D migratory mechanism not mimicked by 2D matrices.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA. adoyle@mail.nih.gov

ABSTRACT
Current concepts of cell migration were established in regular two-dimensional (2D) cell culture, but the roles of topography are poorly understood for cells migrating in an oriented 3D fibrillar extracellular matrix (ECM). We use a novel micropatterning technique termed microphotopatterning (microPP) to identify functions for 1D fibrillar patterns in 3D cell migration. In striking contrast to 2D, cell migration in both 1D and 3D is rapid, uniaxial, independent of ECM ligand density, and dependent on myosin II contractility and microtubules (MTs). 1D and 3D migration are also characterized by an anterior MT bundle with a posterior centrosome. We propose that cells migrate rapidly through 3D fibrillar matrices by a 1D migratory mechanism not mimicked by 2D matrices.

Show MeSH
Related in: MedlinePlus