Claudin-1 contributes to the epithelial barrier function in MDCK cells

doi:10.1016/S0171-9335(99)80086-7

European Journal of Cell Biology

Volume 78, Issue 12, December 1999, Pages 849-855

https://doi.org/10.1016/S0171-9335(99)80086-7 Get rights and content

Abstract

Tight junctions (TJs) create a paracellular permeability barrier and also act as a fence preventing intermixing of proteins and lipids between the apical and basolateral plasma membranes. Recently, claudin-1 has been identified as an integral membrane protein localizing at TJs, and introduced claudin-1 can form TJ-like networks in fibroblasts. To investigate the function of claudin-1, MOCK cells were transfected with a mammalian expression vector containing myc-tagged mouse claudin-1, and four stable clones were obtained. The myc-tagged claudin-1 precisely colocalized with both occludin and ZO-1 at cell-cell contact sites, indicating that exogenous claudin-1 was properly targeted to the TJs. Immunoblot analysis revealed that overexpression of claudin-1 increased expression of ZO-1 but not of occludin or ZO-2. The barrier functions of these cells were evaluated by transepithelial electrical resistance (TER) and paracellular flux. Claudin-1-expressing cells exhibited about four times higher TER than wild-type MOCK cells. Consistent with the increase of TER, the cells overexpressing claudin-1 showed reduced paracellular flux, estimated at 4 and 40 kO FITC-dextrans. The.se results suggest that claudin-1 is involved in the barrier function at TJs.

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²: Dr. Tetsuichiro Inai, Department of Developmental Molecular Anatomy, Graduate School of Medical Science, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582/Japan, Fax: ++926426202.

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Claudin-1 contributes to the epithelial barrier function in MDCK cells

Abstract

Anal. Biochem

J. Biol. Chem

J. Biol. Chem

Biochem. Biophys. Res. Commun

Int. Rev. Cytoi

J. Biol. Chem

Functional dissociation of paracellular permeability and transepithelial electrical resistance and disruption of the apical-basolateral intramembrane diffusion barrier by expression of a mutant tight junction membrane protein

J. Cell Biol

COOH terminus of occludin is required for tight junction barrier function in early Xenopus embryos

J. Cell Biol

Junctional complexes in various epithelia

J. Cell Biol

Freeze-fracture replica electron microscopy combined with SDS digestion for cytochemical labeling of integral membrane proteins

Application to the immunogold labeling of intercellular junctional complexes. J. Cell Sci

Occludin: a novel integral membrane protein localizing at tight junctions

J. Cell Biol

Direct association of occludin with ZO-1 and its possible involvement in the localization of occludin at tight junctions

J. Cell Biol

Overexpression of occludin, a tight junction-associated integral membrane protein, induces the formation of intracellular multilamellar bodies bearing tight junction-like structures

J. Cell Sci

Claudin-1 and -2: novel integral membrane proteins localizing at tight junctions with no sequence similarity to occludin

J. Cell Biol

A single gene product, claudin-1 or -2, reconstitutes tight junction strands and recruits occludin in fibroblasts

J. Cell Biol

Structure, biochemistry, and assembly of epithelial tight junctions

Am. J. Physiol

Identification of a 160-kDa polypeptide that binds to the tight junction protein ZO-1

Proc. Natl. Acad. Sci. USA

Breaking through the tight junction barrier

J. Cell Biol