Distribution of α6 and β4 integrins following epithelial abrasion in the rabbit cornea

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Abstract
Integrin complex alpha 6 beta 4 is a component of the hemidesmosome. In the unwounded cornea both the integrin subunits face the laminin-containing basement membrane, but the alpha 6 subunit is also located between the basal cells. While the migrating epithelium is known to be without hemidesmosomes, we investigated the distribution of alpha 6 beta 4 during epithelial healing. Epithelial abrasion 7.5 mm in diameter was mechanically created. The rabbits were killed 1-24 h or 2, 3, or 7 days later. Monoclonal antibodies against alpha 6, beta 4, and laminin A were used to detect their distributions by immunohistochemistry. Positive immunostaining for laminin A on the surface of the unepithelialized stroma indicated that basement membrane was intact after the epithelial abrasion. Three hours after corneal wounding, alpha 6, was detectable around the entire cell up to the leading edge of the migrating epithelium. In the peripheral wound, alpha 6, was also prominently present around the basal and suprabasal cells with only the superficial cell layers being negative. The beta 4 subunit showed a dissimilar distribution; it was not detectable subjacent to the leading edge. After 1 h the immunoreaction for the beta 4 subunit had faded 15 - 20 microns peripheral to the wound margin. Thereafter the subepithelial band was segmentally reassembled, starting from the periphery and progressing toward the central area of the wound. One week after epithelial wounding, immunolabeling for both integrin subunits was indistinguishable from that of the control cornea. Our results indicate that in addition to the beta 4 in the rabbit cornea, the alpha 6 subunit is also complex with another beta subunit during the epithelial healing phase. The results also suggest that basal cells 15 - 20 microns peripheral to the wound margin disassemble their HDs prior to the migration process.