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Ultrastructural Localization of Integrin Subunits β4 and 3 Within the Migrating Epithelial Tongue of In Vivo Human Wounds

Robert A. Underwood, William G. Carter, Marcia L. Usui and John E. Olerud

Department of Medicine (Dermatology) (RAU,MLU,JEO) and Department of Pathobiology (WGC), University of Washington, Seattle, Washington, and Fred Hutchinson Cancer Research Center, Seattle, Washington (WGC)

Correspondence to: Robert Underwood, University of Washington, Department of Medicine (Dermatology), Box 356524, Seattle, WA 98195-6524. E-mail: underwoo{at}u.washington.edu

Subsequent to wounding, keratinocytes must quickly restore barrier function. In vitro wound models have served to elucidate mechanisms of epithelial closure and key roles for integrins 6β4 and 3β1. To extrapolate in vitro data to in vivo human tissues, we used ultrathin cryomicrotomy to simultaneously observe tissue ultrastructure and immunogold localization in unwounded skin and acute human cutaneous wounds. Localization of the β4 integrin subunit in unwounded skin shows dominant hemidesmosomal association and minor basal keratinocyte lateral filopodic cell–cell expression. After wounding, β4 dominantly localized to cytokeratin-rich regions (trailing edge hemidesmosomes) and minor association with lamellipodia (leading edge). β4 colocalizes with 3 within filopodia juxtaposed to wound matrix, and increased concentrations of β4 were found in cytoplasmic vesicles within basal keratinocytes of the migrating tongue. 3 integrin subunit dominantly localized to filopodia within basal keratinocyte lateral cell–cell interfaces in unwounded skin and both cell–cell and cell–matrix filopodic interactions in wounded skin. This study indicates that β4 interacts with the extracellular environment through both stable and transient interactions and may be managed through a different endosomal trafficking pathway than 3. 3 integrin, despite its ability to respond to alternate ligands after wounding, does so through a single structure, the filopodia. (J Histochem Cytochem 57:123–142, 2009)

Key Words: human • wound • healing • skin • integrin • keratinocyte • epithelial • migration • 6β4 • 3β1

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