title: Ultrastructural localization of integrin subunits beta4 and alpha3 within the migrating epithelial tongue of in vivo human wounds.
abstract: 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 alpha6beta4 and alpha3beta1. 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 beta4 integrin subunit in unwounded skin shows dominant hemidesmosomal association and minor basal keratinocyte lateral filopodic cell-cell expression. After wounding, beta4 dominantly localized to cytokeratin-rich regions (trailing edge hemidesmosomes) and minor association with lamellipodia (leading edge). beta4 colocalizes with alpha3 within filopodia juxtaposed to wound matrix, and increased concentrations of beta4 were found in cytoplasmic vesicles within basal keratinocytes of the migrating tongue. alpha3 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 beta4 interacts with the extracellular environment through both stable and transient interactions and may be managed through a different endosomal trafficking pathway than alpha3. alpha3 integrin, despite its ability to respond to alternate ligands after wounding, does so through a single structure, the filopodia.
authors.affiliation: University of Washington, Department of Medicine (Dermatology), Seattle, WA 98195-6524, USA. underwoo@u.washington.edu
authors.forename: Robert A
authors.initials: RA
authors.lastname: Underwood
authors.forename: William G
authors.initials: WG
authors.lastname: Carter
authors.forename: Marcia L
authors.initials: ML
authors.lastname: Usui
authors.forename: John E
authors.initials: JE
authors.lastname: Olerud
mesh_terms: D002465:Cell Movement; D003593:Cytoplasm / Q000378:metabolism; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005629:Frozen Sections; D022002:Hemidesmosomes / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D039422:Integrin alpha3 / Q000378:metabolism*; D039663:Integrin beta4 / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D011554:Pseudopodia / Q000378:metabolism; D012867:Skin / Q000293:injuries* / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D039422:Integrin alpha3; D039663:Integrin beta4; D021122:Protein Subunits; D007633:Keratins