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104. Jahrestagung der Deutschen Ophthalmologischen Gesellschaft 2006
Abstract
Abstract
FR.09.02 Cell-matrix interaction in podosomal structures of human corneal epithelial cells Kampik D., Geerling G., Grehn F., Schlunck G.
Universität Würzburg, Augenklinik und Poliklinik Objective: Cell-matrix interaction deficiencies in corneal epithelial cells can lead to recurrent erosions, dystrophic corneal diseases and improper healing of superficial wounds. We have previously detected podosome-like adhesive structures in primary human corneal epithelial cells. Here, we report on cell-matrix interactions in podosome-like adhesions.
Methods: Primary human corneal epithelial cells (hCEC) and an immortalized corneal epithelial cell line were plated on fibronectin-coated coverslips. Cells were treated with phorbol-12,13-dibutyrate (PDBu) or vehicle, fixed and triple-stained with phalloidin-TRITC or antibodies to detect F-actin, cortactin and vinculin, respectively. To assess matrix remodelling, cells were plated on FITC-tagged fibronectin for appropriate times, stained for F-actin and cortactin and viewed with a confocal laser-scanning microscope.
Results: Untreated primary hCEC as well as a corneal epithelial cell line express podosomal structures as detected by basal colocalization of F-actin and cortactin. PDBu induced podosomal structures in both cell types, induction was more pronounced in primary hCEC. Fibronectin is degraded at podosomal sites and accumulates at focal adhesions.
Conclusions: Podosome-like structures can be studied in corneal epithelial cell lines. The role of extracellular matrix degradation at podosome-like sites in cell adhesion and migration deserves further attention.
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