Purpose: To determine whether production and localization of beta 4 integrin is altered during in vivo corneal epithelial cell migration in response to debridement wounding.
Methods: Rat corneas were wounded and animals were killed at times ranging from 3 hours to 14 days. At various time points, corneal epithelial integrins were quantitated by gel electrophoresis and immunoblotting of epithelial extracts and then were localized by immunohistochemistry.
Results: As early as 6 hours after wounding, an increase in the amount of the beta 4 integrin subunit expressed per microgram of total protein was observed. The level of beta 4 continued to increase until wound closure. By 14 days after wounding, beta 4 expression returned to control levels. The level of expression of beta 1 and alpha (v) integrins were found not to change significantly throughout migration. Immunohistochemical analyses using antibodies against either the beta 4 integrin subunit or HD1, a hemidesmosomal plaque component, showed that in control sections, beta 4 integrin and HD1 codistributed in a linear staining pattern above the basement membrane. As early as 4 hours after wounding, beta 4 was present in both basal and suprabasal epithelial cells, and HD1 was retained at the basal aspect of the epithelial basal cells.
Conclusions: These data show that changes in expression and localization of beta 4 integrin occur in the corneal epithelium in response to debridement wounding in vivo. Previously, we had shown that quantitative changes in beta 4 integrin expression do not occur in an in vitro organ culture model used for the study of corneal epithelial cell migration. Increased beta 4 expression may not be required for migration per se, but it may be play a role in either stabilizing cell:cell or cell:substrate adhesion in vivo or in preparing cells to undergo mitosis during restratification.