| Description |
Author Affiliation: Lake J ( CUO-Recherche Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 2Médecine Régénératrice, Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 3Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Québe.); Zaniolo K ( CUO-Recherche Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 2Médecine Régénératrice, Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 3Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Québe.); Gingras MÈ ( CUO-Recherche Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada.); Couture C ( CUO-Recherche Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 2Médecine Régénératrice, Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 3Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Québe.); Salesse C ( CUO-Recherche Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 2Médecine Régénératrice, Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 3Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Québe.); Guérin SL ( CUO-Recherche Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 2Médecine Régénératrice, Centre de Recherche FRQS du CHU de Québec, Québec, Québec, Canada 3Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Québe.) |
| Abstract |
PURPOSE: The early step of corneal wound healing is characterized by the massive production of fibronectin (FN), whose secretion is progressively replaced by collagens from the basal membrane as wound healing proceeds. Here, we examined whether expression of the gene encoding the 5 subunit from the FN-binding integrin 5ß1 changes as corneal epithelial cells (CECs) are cultured in the presence of collagen type I (CI) or type IV (CIV). METHODS: Responsiveness of the 5 gene toward collagen was determined by transfection of 5 promoter/chloramphenicol acetyltransferase (CAT) plasmids into rabbit and human CECs cultured on BSA or collagens. Electrophoretic mobility shift assays and Western blots were used to monitor the transcription factors required for basal 5 gene transcription in the presence of collagens. Gene profiling on microarrays was used to determine the impact of collagens on the patterns of genes expressed by CECs. RESULTS: All collagen types repressed the full-length 5/CAT promoter activity in confluent CECs. A moderate increase was observed in subconfluent rabbit CECs grown on CIV but not on CI. These collagen-dependent regulatory influences also correlated with alterations in the transcription factors Sp1/Sp3, NFI, and AP-1 that ensure 5 gene basal transcription. Microarray analyses revealed that CI more profoundly altered the pattern of genes expressed by human CECs than CIV. CONCLUSIONS: Collagens considerably suppressed 5 gene expression in CECs, suggesting that during wound healing, they may interfere with the influence FN exerts on CECs by altering their adhesive and migratory properties through a mechanism involving a reduction in 5 gene expression. |
| Subject Keyword |
Collagen Metabolism Corneal Injuries Genetics Epithelium, Corneal Gene Expression Regulation Integrin Alpha5 Animals Blotting, Western Cells, Cultured Pathology Electrophoretic Mobility Shift Assay Injuries Biosynthesis Promoter Regions, Genetic Rabbits Reverse Transcriptase Polymerase Chain Reaction Research Support, Non-u.s. Gov't Discipline Ophthalmology |
