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Published online 16 April 2001. doi:10.1083/jcb.153.2.263
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© The Rockefeller University Press, 0021-9525/2001//263 $5.00
The Journal of Cell Biology, Volume 153, Number 2, , 2001 263-272

Original Article

Conversion of Zonulae Occludentes from Tight to Leaky Strand Type by Introducing Claudin-2 into Madin-Darby Canine Kidney I Cells



Mikio Furusea, Kyoko Furuseb, Hiroyuki Sasakib, and Shoichiro Tsukitaa

a Department of Cell Biology, Faculty of Medicine, Kyoto University, Kyoto 606-8501, Japan
b KAN Research Institute Inc., Kyoto 600-8317, Japan
Department of Cell Biology, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan.81-75-753-466081-75-753-4372

htsukita{at}mfour.med.kyoto-u.ac.jp

There are two strains of MDCK cells, MDCK I and II. MDCK I cells show much higher transepithelial electric resistance (TER) than MDCK II cells, although they bear similar numbers of tight junction (TJ) strands. We examined the expression pattern of claudins, the major components of TJ strands, in these cells: claudin-1 and -4 were expressed both in MDCK I and II cells, whereas the expression of claudin-2 was restricted to MDCK II cells. The dog claudin-2 cDNA was then introduced into MDCK I cells to mimic the claudin expression pattern of MDCK II cells. Interestingly, the TER values of MDCK I clones stably expressing claudin-2 (dCL2-MDCK I) fell to the levels of MDCK II cells (>20-fold decrease). In contrast, when dog claudin-3 was introduced into MDCK I cells, no change was detected in their TER. Similar results were obtained in mouse epithelial cells, Eph4. Morphometric analyses identified no significant differences in the density of TJs or in the number of TJ strands between dCL2-MDCK I and control MDCK I cells. These findings indicated that the addition of claudin-2 markedly decreased the tightness of individual claudin-1/4–based TJ strands, leading to the speculation that the combination and mixing ratios of claudin species determine the barrier properties of individual TJ strands.

Key Words: tight junction • zonula occludens • claudin • Madin-Darby canine kidney cells • transepithelial electric resistance

© 2001 The Rockefeller University Press

Abbreviations used in this paper: E-face, extracellular face; P-face, protoplasmic face; TER, transepithelial electric resistance; TJ, tight junction.


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