Connexin 26 expression prevents down-regulation of barrier and fence functions of tight junctions by Na+/K+-ATPase inhibitor ouabain in human airway epithelial cell line Calu-3

Mitsuru Go, Takashi Kojima, Ken ichi Takano, Masaki Murata, Junichi Koizumi, Makoto Kurose, Ryuta Kamekura, Makoto Osanai, Hideki Chiba, David C. Spray, Tetsuo Himi, Norimasa Sawada

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Gap junctions are considered to play a crucial role in differentiation of epithelial cells and to be associated with tight junction proteins. In this study, to investigate the role of gap junctions in regulation of the barrier function and fence function on the tight junctions, we introduced the Cx26 gene into human airway epithelial cell line Clau-3 and used a disruption model of tight junctions employing the Na+/K+-ATPase inhibitor ouabain. In parental Calu-3 cells, gap junction proteins Cx32 and Cx43, but not Cx26, and tight junction proteins occludin, JAM-1, ZO-1, claudin-1, -2, -3, -4, -5, -6, -7, -8, -9, and -14 were detected by RT-PCR. The barrier function and fence function of tight junctions were well maintained, whereas the GJIC was low level. Treatment with ouabain caused disruption of the barrier function and fence function of tight junctions together with down-regulation of occludin, JAM-1, claudin-2, and -4 and up-regulation of ZO-1 and claudin-14. In Cx26 transfectants, Cx26 protein was detected by Western blotting and immunocytochemistry, and many gap junction plaques were observed with well-developed tight junction strands. Expression of claudin-14 was significantly increased in Cx26 transfectants compared to parental cells, and in some cells, Cx26 was co-localized with claudin-14. Interestingly, transfection with Cx26 prevented disruption of both functions of tight junctions by treatment with ouabain without changes in the tight junction proteins. Pretreatment with the GJIC blockers 18β-glycyrrhetinic acid and oleamide did not affect the changes induced by Cx26 transfection. These results suggest that Cx26 expression, but not the mediated intercellular communication, may regulate tight junction barrier and fence functions in human airway epithelial cell line Calu-3.

Original languageEnglish (US)
Pages (from-to)3847-3856
Number of pages10
JournalExperimental Cell Research
Volume312
Issue number19
DOIs
StatePublished - Nov 15 2006

Fingerprint

Tight Junctions
Ouabain
Down-Regulation
Epithelial Cells
Cell Line
Tight Junction Proteins
Gap Junctions
Claudin-2
Occludin
Transfection
Claudin-4
Claudin-1
Glycyrrhetinic Acid
Connexin 43
Connexins
Intercellular Junctions
Connexin 26
ATPase inhibitory protein
Up-Regulation
Western Blotting

Keywords

  • Barrier function
  • Cx26
  • Fence function
  • Ouabain
  • Tight junctions

ASJC Scopus subject areas

  • Cell Biology

Cite this

Connexin 26 expression prevents down-regulation of barrier and fence functions of tight junctions by Na+/K+-ATPase inhibitor ouabain in human airway epithelial cell line Calu-3. / Go, Mitsuru; Kojima, Takashi; Takano, Ken ichi; Murata, Masaki; Koizumi, Junichi; Kurose, Makoto; Kamekura, Ryuta; Osanai, Makoto; Chiba, Hideki; Spray, David C.; Himi, Tetsuo; Sawada, Norimasa.

In: Experimental Cell Research, Vol. 312, No. 19, 15.11.2006, p. 3847-3856.

Research output: Contribution to journalArticle

Go, Mitsuru ; Kojima, Takashi ; Takano, Ken ichi ; Murata, Masaki ; Koizumi, Junichi ; Kurose, Makoto ; Kamekura, Ryuta ; Osanai, Makoto ; Chiba, Hideki ; Spray, David C. ; Himi, Tetsuo ; Sawada, Norimasa. / Connexin 26 expression prevents down-regulation of barrier and fence functions of tight junctions by Na+/K+-ATPase inhibitor ouabain in human airway epithelial cell line Calu-3. In: Experimental Cell Research. 2006 ; Vol. 312, No. 19. pp. 3847-3856.
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AU - Kurose, Makoto

AU - Kamekura, Ryuta

AU - Osanai, Makoto

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