Effect of RNA interference of tight junction-related molecules on intercellular barrier function in cultured human keratinocytes.

Abstract:

:Accumulating evidence shows that tight junctions (TJs) in the granular layer contribute to the epidermal barrier, suggesting that the regulation of TJ assembly in keratinocytes may provide a clue to understanding the role of barrier formation in epidermis. In this study, we investigated the behavior of TJ-related molecules in cultured human keratinocytes during keratinization induced by transfer to high-calcium medium, and the effect of RNA interference of TJ-related molecules on intercellular permeability and morphological features. The expression of TJ-related molecules and transepithelial electrical resistance were increased by transfer to high-calcium medium. In cells under the same conditions, we observed by freeze-fracture electron microscopy that TJ strands developed on the apposing cell membranes. In contrast, the transepithelial electrical resistance was clearly suppressed when the expression of claudin-1 or occludin was blocked by RNA interference. The morphological features of these knock-down cells were the same as those of MOCK cells, except for a marked decrease in the number of TJ strands. Furthermore, claudin-1 suppression inhibited occludin localization at the cell membrane, whereas suppression of occludin did not influence the localization of claudin-1. These results suggest that claudin-1 plays a crucial role in recruiting occludin to TJs, and that occludin is involved in intercellular barrier function.

journal_name

Arch Dermatol Res

authors

Yamamoto T,Saeki Y,Kurasawa M,Kuroda S,Arase S,Sasaki H

doi

10.1007/s00403-008-0868-8

subject

Has Abstract

pub_date

2008-10-01 00:00:00

pages

517-24

issue

9

eissn

0340-3696

issn

1432-069X

journal_volume

300

pub_type

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