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Tight junction protein MAGI-1 is up-regulated by transfection with connexin 32 in an immortalized mouse hepatic cell line: cDNA microarray analysis

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Abstract

Gap junctions are considered to play a crucial role in differentiation of epithelial cells, including hepatocytes. Recently, we found that Cx32 but not Cx26 was closely related to tight junctional proteins in primary cultured rat hepatocytes (Kojima et al., Exp Cell Res 263:193–201, 2001) and that Cx32 formation and/or Cx32-mediated intercellular communication could induce expression and function of tight junctions in a mouse hepatic cell line (Kojima et al., Exp Cell Res 276:40–51, 2002). In this study, to investigate the mechanisms of induction of tight junctions by transfection with Cx32, we performed cDNA microarray analysis of Cx32 transfectants, compared with parental cells derived from Cx32-deficient hepatocytes. In cDNA microarray analysis, a 2.5-fold increase in expression of membrane-associated guanylate kinase with inverted orientation-1 (MAGI-1), which is known to be localized at adherens and tight junction regions, was observed. High expression of MAGI-1 in Cx32 transfectants was confirmed by Western blotting and RT-PCR. MAGI-1 was colocalized with occludin, claudin-2, ZO-1, and F-actin, but not with E-cadherin in the apical-most regions at cell borders of Cx32 transfectants, similar to junctional adhesion molecule-1 (JAM-1), which may play a crucial role in formation and assembly of tight junctions. Treatment with the gap junction blocker 18β-glycyrrhetinic acid did not affect expression of MAGI-1 and JAM-1 in Cx32 transfectants. These results suggest that Cx32 expression is in part related to induction of tight junctions through modulation of MAGI-1 expression in an immortalized mouse hepatic cell line.

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Acknowledgements

We are grateful to Dr Y. Hata (Tokyo Medical and Dental University) for the MAGI-1 antibody and Dr T. Kita (Kyoto University) for the JAM-1 antibody.

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Authors and Affiliations

  1. Department of Pathology, Sapporo Medical University School of Medicine, Sapporo, Japan

    Masaki Murata, Takashi Kojima, Toshinobu Yamamoto, Mitsuru Go, Ken-ichi Takano, Hideki Chiba & Norimasa Sawada

  2. Department of Molecular Biology, Cancer Research Institute, Sapporo Medical University School of Medicine, Sapporo, Japan

    Takashi Tokino

Authors
  1. Masaki Murata
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  2. Takashi Kojima
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  3. Toshinobu Yamamoto
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  4. Mitsuru Go
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  5. Ken-ichi Takano
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  6. Hideki Chiba
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  7. Takashi Tokino
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  8. Norimasa Sawada
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Corresponding author

Correspondence to Takashi Kojima.

Additional information

This work was supported in by Grants-in-Aid from the Ministry of Education, Culture, Sports, Science Technology and the Ministry of Health, Labour and Welfare of Japan, and by the Kato Memorial Bioscience Foundation, the Uehara Memorial Foundation, the Suhara Memorial Foundation, the Smoking Research Foundation, and the Long-Range Research Initiative Project of the Japan Chemical Industry Association.

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Murata, M., Kojima, T., Yamamoto, T. et al. Tight junction protein MAGI-1 is up-regulated by transfection with connexin 32 in an immortalized mouse hepatic cell line: cDNA microarray analysis. Cell Tissue Res 319, 341–347 (2005). https://doi.org/10.1007/s00441-004-1017-0

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