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DNA repair after DNA fragmentation in mouse small intestinal epithelial cells

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Abstract

In our earlier work, we found that, in mice, i.p. injection of anti-CD3 monoclonal antibody activated intraepithelial lymphocytes (iIEL), leading to DNA fragmentation in villous epithelial cells of the duodenum and jejunum within 30 min. By 2 h after injection, nearly half of the enterocytes had detached from the villi, and DNA fragmentation could barely be detected in the remaining villous epithelium. We hypothesized that DNA had been repaired in enterocytes in which DNA fragmentation had previously been induced. In this study, enterocytes became negative for TUNEL staining at 60 min after anti-CD3 treatment, prior to detachment. The remaining villous epithelial cells, after DNA fragmentation and detachment, were found to be positive for 5-bromo-2-deoxyuridine labeling. To confirm whether fragmented DNA had been repaired in situ, we investigated the appearance and/or mobilization of DNA-repair-related proteins. Focus formation, a typical staining pattern of repair-related proteins including phosphorylated H2AX, phospo-ATM substrate, and Nbs1, was observed 30 min after anti-CD3 injection, with the kinetics virtually identical to that of DNA fragmentation. The co-localization of γ-H2AX and phospo-ATM substrate was also confirmed. The disappearance of a positive reaction for TUNEL staining in previously fragmented DNA, the appearance of representative DNA-repair-related proteins, the coincidence of the kinetics of DNA fragmentation and this appearance of DNA-repair-related proteins, and the co-localization of two of the repair-related proteins strongly indicated that enterocyte DNA could be repaired after it had been fragmented in vivo. Thus, DNA fragmentation per se may not necessarily be an immediate sign of cell death.

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Authors and Affiliations

  1. Division of Immunology and Embryology, Department of Cell Biology, Tohoku University School of Medicine, 980–8575, Sendai, Japan

    Masaki Ogata, Takehiko Oomori, Hiroyuki Soga, Yuta Ota, Ari Itoh, Takaji Matsutani, Masanobu Nanno, Ryuji Suzuki & Tsunetoshi Itoh

  2. Faculty of Health Sciences, Kinjo University, Hakusan, Japan

    Hiroyuki Soga

  3. Department of Applied Research, Division of Immunology, Yakult Central Institute for Microbiological Research, Tokyo, Japan

    Masanobu Nanno

  4. Department of Rheumatology & Clinical Immunology, Clinical Research Center for Allergy & Rheumatology, National Sagamihara Hospital, Sagamihara, Japan

    Ryuji Suzuki

Authors
  1. Masaki Ogata
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  2. Takehiko Oomori
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  3. Hiroyuki Soga
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  4. Yuta Ota
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  5. Ari Itoh
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  6. Takaji Matsutani
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  7. Masanobu Nanno
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  8. Ryuji Suzuki
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  9. Tsunetoshi Itoh
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Corresponding author

Correspondence to Masaki Ogata.

Additional information

This work was supported in part by a Grant-in-aid for Scientific Research from the Ministry of Education, Science and Culture, Japan (16590132 to T.M., 16390045 to T.I., and 20590181 to M.O.).

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Ogata, M., Oomori, T., Soga, H. et al. DNA repair after DNA fragmentation in mouse small intestinal epithelial cells. Cell Tissue Res 335, 371–382 (2009). https://doi.org/10.1007/s00441-008-0727-0

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  • DOI: https://doi.org/10.1007/s00441-008-0727-0

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