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(Investigative Ophthalmology and Visual Science. 2003;44:1355-1363.)
© 2003 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.02-0046
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Tractional Force Generation by Human Müller Cells: Growth Factor Responsiveness and Integrin Receptor Involvement

Clyde Guidry, Kelley M. Bradley, and Jeffery L. King

From the Department of Ophthalmology, University of Alabama at Birmingham, Birmingham, Alabama.

PURPOSE. To assess the ability of human Müller cells to generate tractional forces and to determine the role of growth factors and collagen binding integrins in this process.

METHODS. Müller cells were isolated from papain-DNase–digested human retina. Cell identity and changes in cell phenotype were confirmed by immunodetection of glial fibrillary acidic protein (GFAP), cellular retinaldehyde-binding protein (CRALBP), vimentin, and {alpha}-smooth muscle actin ({alpha}-SMA). Generation of tractional force was assessed with a tissue culture assay involving incubation of cells on three-dimensional collagen gels. The effects of contraction-promoting growth factors were examined by adding these directly to the culture medium. Müller cell expression and the involvement of specific integrin receptors were assessed by immunodetection, RT-PCR, and subunit-specific blocking antibodies.

RESULTS. During maintenance in culture, human Müller cells adopted a fibroblast-like phenotype capable of generating tractional forces. Matrix contraction was stimulated in a dose-dependent fashion by insulin-like growth factor I and platelet-derived growth factor. Modest responses were observed with high concentrations of transforming growth factor (TGF)-ß1 and -ß2. Müller cells express all four integrin subunits that comprise the collagen-binding receptors including {alpha}1, {alpha}2, {alpha}3, and ß1. Blocking antibodies against receptor subunits {alpha}2 and ß1 significantly reduced the overall rate of matrix contraction. Antibodies against the {alpha}1 subunit were modestly inhibitory, whereas anti-{alpha}3 was without effect.

CONCLUSIONS. Human Müller cells acquire the capacity to generate tractional forces in vitro and the contraction-promoting growth factors insulin-like growth factor (IGF)-I and platelet-derived growth factor (PDGF) are potent stimuli. Generation of tractional force by Müller cells primarily involves integrin receptors containing {alpha}2 and ß1 subunits.




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