Investigation of trans-interaction between claudins, tight junction constitutive transmembrane proteins, and paracellular permeability.

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We established HEK293 cells, which does not form tight junctions (TJs), expressing RFP-cldn-1, RFP-cldn-10b, EGFP-cldn-10a or EGFP-cldn-10b. EGFP-cldn-10a- and RFP-cldn-1-expressing cells, EGFP-cldn-10b- and RFP-cldn-1-expressing cells, or EGFP-cldn-10a- and RFP-cldn-10b-expressing cells were co-cultured and examined by confocal laser scanning microscopy. Both EGFP-cldn-10a and EGFP-cldn-10b were not co-localized with RFP-cldn-1 in cell-cell contacts between heterotypic cells. In contrast, EGFP-cldn-10a was co-localized with RFP-cldn-10b. Using freeze-fracture electron microscopy, P-face-associated or E- face-associated TJ particles were observed in EGFP-cldn-10a- or EGFP-cldn-10b-expressing cells, respectively. When cldn-10a or cldn-10b was expressed in TJ-bearing MDCK I cells, cldn-10a did not alter trans-epithelial electrical resistance (TER), but cldn-10b markedly reduced TER.