Colocalization of laminin and fibronectin in bovine lens epithelial cells in vitro.
The distribution and organization of the extracellular matrix (ECM) proteins laminin, fibronectin, entactin, and type IV collagen were investigated in primary colonies and secondary cultures of bovine lens epithelial cells using species-specific antisera and indirect immunofluorescence microscopy. Primary cell colonies fixed in formaldehyde and permeabilized with Triton X-100 displayed diffuse intracellular patterns for type IV collagen and entactin, observed to a similar degree throughout the colonies. In contrast, thick bundles of laminin and fibronectin were located on the basal cell surfaces and in between cells in the densely packed center of the colonies, and as "adhesive plaques" and fine extracellular matrix cords in the sparsely populated (migratory) outer edge of the colonies. The distribution of ECM proteins observed in secondary lens epithelial cell cultures was similar to that observed at the periphery of the primary colony. Extraction of the secondary cell cultures with sodium deoxycholate confirmed that laminin and fibronectin were deposited on the basal cell surface. Indeed, the patterns of laminin and fibronectin deposition suggested that these proteins codistribute. These results establish that lens epithelial cells in culture can be used as a model system to study the synthesis and extracellular deposition of the basement membrane proteins, laminin and fibronectin.
Cammarata
P R
PR
Department of Anatomy, Texas College of Osteopathic Medicine, Fort Worth 76107.
Smith
J Y
JY
D000818:Animals; D001485:Basement Membrane / Q000166:cytology; D002417:Cattle; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis; D004847:Epithelial Cells; D005109:Extracellular Matrix / Q000648:ultrastructure; D005353:Fibronectins / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000032:analysis; D007797:Laminin / Q000032:analysis*; D007908:Lens, Crystalline / Q000166:cytology*; D008562:Membrane Glycoproteins
D005353:Fibronectins; D006023:Glycoproteins; D007797:Laminin; D008562:Membrane Glycoproteins; C029967:nidogen; D003094:Collagen