Interkeratinocyte adherens junctions: Immunocytochemical visualization of cell-cell junctional structures, distinct from desmosomes, in human epidermis

Abstract

Vinculin and β-catenin are intracellular attachment proteins linking transmembrane adhesion molecules (E-cadherin) to the actin microfilament cytoskeleton, thus participating in formation of cell-cell adherens junctions, or zonulae adherentes. This type of junction was only recently described in human epidermis due to the imprecise morphological criteria for its recognition, In this study, we investigated the relationship between the expression of the zonula adherens-associated proteins vinculin, β-catenin, E-cadherin, and actin, on the one hand, and the presence of electron microscopically discernable structures in normal human epidermis on the other. Mouse jejunal epithelium with its orderly arrangement of various junctional structures sewed as a positive control. Simple and dual post-embedding immunogold labeling was performed on ultrathin sections of Lowicryl K4M and Lowicryl K11M embedded tissues. The overall distribution of the antigens in human epidermis was evaluated on frozen tissue sections using immunofluorescence and laser confocal scanning microscopy. Antibodies against proteins associated with desmosomes (i.e., keratins, desmoglein 1, and plakoglobin) were used as controls. Vinculin and β-catenin were localized to junctional structures distinct from desmosomes, thus defining the presence zonulae adherentes. Labeling of actin and E-cadherin was less clearly restricted to the junctions, but these two proteins were also co-expressed at zonulae adherentes and not at desmosomes. In human epidermis, zonula adherens-associated labeling was consistently detected near desmosomes, indicating the possibility of a functional relationship between the two types of junctions.