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Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires ?₅?₁ integrin engagement.

ABSTRACT: Despite the importance of matrix rigidity on cell functions, many aspects of the mechanosensing process in highly migratory cells remain elusive. Here, we studied the migration of highly motile keratocytes on culture substrates with similar biochemical properties and rigidities spanning the range between soft tissues (~kPa) and stiff culture substrates (~GPa). We show that morphology, polarization and persistence of motile keratocytes are regulated by the matrix stiffness over seven orders of magnitude, without changing the cell spreading area. Increasing the matrix rigidity leads to more F-actin in the lamellipodia and to the formation of mature contractile actomyosin fibers that control the cell rear retraction. Keratocytes remain rounded and form nascent adhesions on compliant substrates, whereas large and uniformly distributed focal adhesions are formed on fan-shaped keratocytes migrating on rigid surfaces. By combining poly-L-lysine, fibronectin and vitronectin coatings with selective blocking of ?_v?₃ or ?₅?₁ integrins, we show that ?_V?₃ integrins permit the spreading of keratocytes but are not sufficient for polarization and rigidity sensing that require the engagement of ?₅?₁ integrins. Our study demonstrates a matrix rigidity-dependent regulation of the directional persistence in motile keratocytes and refines the role of ?_v?₃ and ?₅?₁ integrins in the molecular clutch model.

SUBMITTER: Riaz M

PROVIDER: S-EPMC5039689 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature

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Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires α<sub>5</sub>β<sub>1</sub> integrin engagement.

Riaz Maryam M Versaevel Marie M Mohammed Danahe D Glinel Karine K Gabriele Sylvain S

Scientific reports 20160928

Despite the importance of matrix rigidity on cell functions, many aspects of the mechanosensing process in highly migratory cells remain elusive. Here, we studied the migration of highly motile keratocytes on culture substrates with similar biochemical properties and rigidities spanning the range between soft tissues (~kPa) and stiff culture substrates (~GPa). We show that morphology, polarization and persistence of motile keratocytes are regulated by the matrix stiffness over seven orders of ma ...[more]

PMID: 27678055

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Project description:A tissue engineering approach can provide replacement salivary gland structures to patients with hyposalivation disorders and xerostomia. Salivary human stem/progenitor cells (hS/PCs) were isolated from healthy regions of parotid glands of head and neck surgery patients, expanded, then encapsulated in biocompatible hyaluronate (HA)-based hydrogels. These bioactive hydrogels provide a surrogate territorial matrix suitable for the dynamic assembly, growth and reorganization of salivary gland components. This study examined the dynamics of salivary microstructure formation, growth, and reorganization using time-lapse imaging over 15 h. Immunofluorescence detection monitored production of individual basement membrane components forming around developing microstructures, and Ki67 assessed proliferation. Dynamic movements in hydrogels were quantified by measuring angular velocity (?) of rotating salivary microstructures and changes in basement membrane architecture during microstructure growth. Integrin involvement in the dynamic reassembly was assessed using knockdown and inhibitor approaches. Single hS/PCs expanded over 5 days into spherical microstructures typically containing 3-10 cells. In larger macrostructures, proliferation occurred near the peripheral basement membrane that underwent growth-associated cycles of thinning and collapse. De novo secretion of laminin/collagen IV from reorganizing hS/PCs preceded that of perlecan/HSPG2. Microstructures routinely expressed ?1 integrin-containing complexes at basement membrane-associated regions and exhibited spontaneous and coordinated rotation during basement membrane maturation. ?1 integrin siRNA knockdown at the single-cell state prevented hS/PC microstructure growth. After microstructure formation, ?1 integrin knockdown reduced rotation and mean ? by 84%. Blockade of the ?1 integrin subunit (CD49a) that associates with ?1 reduced mean ? by 66%. Studies presented here show that initial hS/PC structure growth and basement membrane maturation depends on ?1?1-integrin mediated signaling. Coordinated cellular motility during neotissue reorganization reminiscent of salivary gland acini was critically dependent both on hS/PC-secretion of laminin,collagen type-IV, and perlecan/HSPG2 and the force-driven interactions of ?1?1-integrin activation. We conclude that ?1?1-integrin plays a critical role in establishing human salivary gland coordinated structure and function, and that its activation in tissue engineered systems is essential to tissue assembly.

Dataset Information

Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires ?₅?₁ integrin engagement.

Publications

Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires α<sub>5</sub>β<sub>1</sub> integrin engagement.

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Dataset Information

Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires ?5?1 integrin engagement.

Publications

Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires α<sub>5</sub>β<sub>1</sub> integrin engagement.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Persistence of fan-shaped keratocytes is a matrix-rigidity-dependent mechanism that requires ?₅?₁ integrin engagement.