title: Tractional force generation by human müller cells: growth factor responsiveness and integrin receptor involvement.
abstract: OBJECTIVE
  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)-beta1 and -beta2. Müller cells express all four integrin subunits that comprise the collagen-binding receptors including alpha1, alpha2, alpha3, and beta1. Blocking antibodies against receptor subunits alpha2 and beta1 significantly reduced the overall rate of matrix contraction. Antibodies against the alpha1 subunit were modestly inhibitory, whereas anti-alpha3 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 alpha2 and beta1 subunits.
authors.affiliation: Department of Ophthalmology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA. guidry@vision.vsrc.uab.edu
authors.forename: Clyde
authors.initials: C
authors.lastname: Guidry
authors.forename: Kelley M
authors.initials: KM
authors.lastname: Bradley
authors.forename: Jeffery L
authors.initials: JL
authors.lastname: King
mesh_terms: D000199:Actins / Q000378:metabolism; D000328:Adult; D000368:Aged; D018821:CD18 Antigens / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D018929:Cell Culture Techniques; D002469:Cell Separation; D003094:Collagen / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D039421:Integrin alpha2 / Q000378:metabolism*; D008875:Middle Aged; D009474:Neurons / Q000187:drug effects* / Q000502:physiology*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D012160:Retina / Q000166:cytology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016212:Transforming Growth Factor beta / Q000494:pharmacology; D053773:Transforming Growth Factor beta1
chemical_list: C020932:11-cis-retinal-binding protein; D000199:Actins; D018821:CD18 Antigens; D002352:Carrier Proteins; D005904:Glial Fibrillary Acidic Protein; D006133:Growth Substances; D039421:Integrin alpha2; D010982:Platelet-Derived Growth Factor; C508960:TGFB1 protein, human; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D007334:Insulin-Like Growth Factor I; D003094:Collagen