Immunodetection of connexins and cadherins in corneal fibroblasts and myofibroblasts [published erratum appears in Invest Ophthalmol Vis Sci 1996 Nov;37(12):2366]

• Published in: Investigative ophthalmology & visual science Vol. 37; no. 9; pp. 1740 - 1748
• Main Authors: Petridou, S, Masur, SK
• Format: Journal Article
• Language: English
• Published: Rockville, MD ARVO 01.08.1996; Association for Research in Vision and Ophtalmology
• Subjects: alpha Catenin; Amino Acid Sequence; Animals; Antibodies; beta Catenin; Biological and medical sciences; Blotting, Western; Cadherins - analysis; Cadherins - immunology; Cells, Cultured; Connexin 43 - analysis; Connexins - analysis; Connexins - immunology; Cornea - cytology; Cytoskeletal Proteins - analysis; Cytoskeletal Proteins - chemistry; Cytoskeletal Proteins - immunology; Desmoplakins; Eye and associated structures. Visual pathways and centers. Vision; Fibroblasts - cytology; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Molecular Sequence Data; Peptide Fragments - chemistry; Peptide Fragments - immunology; Rabbits; Trans-Activators; Vertebrates: nervous system and sense organs; Cell culture; Cornea; Myofibroblast; Rabbit; Lagomorpha; Cadherin; Eye; Visual system; Connexin; Vertebrata; Mammalia; Microscopy; Animal; Immunoblotting assay; Immunofluorescence; Fibroblast
• ISSN: 0146-0404; 1552-5783

In normal cornea, stromal fibroblasts (keratocytes) interact with one another by gap junctions. After corneal wounding, the remaining corneal stroma cells are phenotypically fibroblasts and myofibroblasts. For insight into the respective roles of fibroblasts and myofibroblasts in wound healing, the authors have investigated the molecular basis of cell-cell interaction in cultures of corneal fibroblasts and corneal myofibroblasts. Using Western blot analysis and immunofluorescent microscopy, the authors determined the relative expression and localization of junction proteins-connexins, cadherins, and cadherin-associated proteins (catenins)-in cultured fibroblasts and myofibroblasts. In cultured corneal fibroblasts, the gap junction protein, connexin 43, was highly expressed and was localized to dense maculae; cadherins were not detected in cell-cell contacts. Cultured myofibroblasts showed the opposite pattern: Cadherins were highly expressed and localized at the cell-cell contacts, whereas myofibroblast connexin 43 was primarily intracellular. Myofibroblast cadherin was identified by a pan-cadherin antibody as a molecule of 135 kDa that reacted weakly with an N-cadherin monoclonal antibody. In addition, cadherin-associated cytoplasmic proteins, alpha- and beta-catenins, co-localized with cadherin at the cell-cell borders of the myofibroblasts. The presence of connexin 43 at the cell-cell borders of corneal fibroblasts is consistent with a primary communication role of junctions in confluent corneal fibroblasts. In contrast, the presence of cadherin at the cell-cel borders of myofibroblasts may provide a site for insertion of actin filaments. A cadherin-actin association could support actin-based force generation for effective wound closure.