title: Beta1 and beta3 integrins cooperate to induce syndecan-4-containing cross-linked actin networks in human trabecular meshwork cells.
abstract: OBJECTIVE
  To characterize the molecular composition of cross-linked actin networks (CLANs) and the regulation of their formation by integrins in normal human trabecular meshwork (TM) cells. CLANs have been observed in steroid-treated and glaucomatous TM cells and have been suggested to contribute to decreased outflow facility by altering the contractility of the TM.
  
  
  METHODS
  Immunofluorescence microscopy was used to identify molecular components of CLANs and quantitate CLAN formation in HTM cells plated on coverslips coated with various extracellular matrix (ECM) proteins (fibronectin, types I and IV collagen, and vitronectin), vascular cell adhesion molecule (VCAM)-1, or activating antibodies against beta1, beta3, or alpha2beta1 integrins. These integrin antibodies were also used as soluble ligands.
  
  
  RESULTS
  CLAN vertices contained the actin-binding proteins alpha-actinin and filamin and the signaling molecules syndecan-4 and PIP2. CLANs lacked Arp3 and cortactin. CLAN formation was dependent on the ECM substrate and was significantly higher on fibronectin and VCAM-1 compared with vitronectin, types I or IV collagen. Adsorbed beta1 integrin antibodies also induced CLANs, whereas adsorbed beta3 or alpha2beta1 integrin antibodies did not. Soluble beta3 integrin antibodies, however, induced CLANs and actually enhanced CLAN formation in cells spread on fibronectin, VCAM-1, type I or type IV collagen, or beta1 integrin antibodies.
  
  
  CONCLUSIONS
  CLANs are unique actin-branched networks whose formation can be regulated by beta1 and beta3 integrin signaling pathways. Thus, integrin-mediated signaling events can modulate the organization of the actin cytoskeleton in TM cells and hence could participate in regulating cytoskeletal events previously demonstrated to be involved in controlling outflow facility.
authors.affiliation: Department of Pathology and Laboratory Medicine, University of Wisconsin, Madison 53706, USA.
authors.forename: Mark S
authors.initials: MS
authors.lastname: Filla
authors.forename: Anne
authors.initials: A
authors.lastname: Woods
authors.forename: Paul L
authors.initials: PL
authors.lastname: Kaufman
authors.forename: Donna M
authors.initials: DM
authors.lastname: Peters
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000185:Actinin / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D018929:Cell Culture Techniques; D003285:Contractile Proteins / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology; D064448:Filamins; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D019012:Integrin beta1 / Q000502:physiology*; D039661:Integrin beta3 / Q000502:physiology*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008840:Microfilament Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism; D011509:Proteoglycans / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D053671:Syndecan-4; D014129:Trabecular Meshwork / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*
chemical_list: D000199:Actins; D003285:Contractile Proteins; D064448:Filamins; D005938:Glucocorticoids; D019012:Integrin beta1; D039661:Integrin beta3; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D011509:Proteoglycans; C508304:SDC4 protein, human; D053671:Syndecan-4; D000185:Actinin; D003907:Dexamethasone