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Journal of Gastroenterology

, Volume 46, Issue 4, pp 456–468 | Cite as

The specific linker phosphorylation of Smad2/3 indicates epithelial stem cells in stomach; particularly increasing in mucosae of Helicobacter-associated gastritis

  • Toshiro FukuiEmail author
  • Masanobu Kishimoto
  • Atsushi Nakajima
  • Masao Yamashina
  • Shinji Nakayama
  • Takeo Kusuda
  • Yutaku Sakaguchi
  • Katsunori Yoshida
  • Kazushige Uchida
  • Akiyoshi Nishio
  • Koichi Matsuzaki
  • Kazuichi Okazaki
Original Article—Alimentary Tract

Abstract

Background

The gastric corpus and antrum are believed to contain epithelial stem cells in the isthmus. However, the lack of useful markers has hindered studies of their origin. We explored whether Smad2/3, phosphorylated at specific linker threonine residues (pSmad2/3L-Thr), could serve as a marker for stem cells.

Methods

Stomachs, small intestines, and colons from Helicobacter felis-infected and noninfected C57BL/6 mice were examined. Double immunofluorescent staining of pSmad2/3L-Thr with Ki67, cytokeratin 8, or doublecortin and calcium/calmodulin-dependent protein kinase-like-1 (DCAMKL1) was performed, and pSmad2/3L-Thr immunostaining-positive cells were counted. After immunofluorescent staining, we stained the same sections with hematoxylin–eosin and observed these cells under a light microscope.

Results

In infected mice, pSmad2/3L-Thr immunostaining-positive cells were significantly increased in the corpus and antrum compared with those of noninfected mice (p < 0.0001). The number of Ki67 immunostaining-positive cells in the corpus and antrum of infected mice was also much greater than in the noninfected mice. Although pSmad2/3L-Thr immunostaining-positive cells were detected among the Ki67 cells, immunohistochemical co-localization of pSmad2/3L-Thr with Ki67 was never observed. pSmad2/3L-Thr immunostaining-positive cells showed immunohistochemical co-localization with cytokeratin 8, but some of them showed co-localization or adjacent localization with DCAMKL1 immunostaining-positive cells. Under a light microscope, pSmad2/3L-Thr immunostaining-positive cells indicated undifferentiated morphological features and were confirmed in the isthmus. In small intestines and colons, pSmad2/3L-Thr immunostaining-positive cells were detected in specific epithelial cells around crypt bases, where the respective putative stem cells are thought to exist.

Conclusions

We have identified the significant expression of pSmad2/3L-Thr in specific epithelial cells of the murine stomach and have suggested these cells to be epithelial stem cells.

Keywords

Linker phosphorylation Smad2/3 Helicobacter-associated gastritis Epithelial stem cells 

Notes

Acknowledgments

This study was supported by Grants-in-Aid for Scientific Research (20790516 and 22790676) from the Japan Society for the Promotion of Science (JSPS) and by Intractable Diseases Health and Labor Sciences Research Grants (to K.O.) from the Ministry of Labor and Welfare of Japan.

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Copyright information

© Springer 2010

Authors and Affiliations

  • Toshiro Fukui
    • 1
    Email author
  • Masanobu Kishimoto
    • 1
  • Atsushi Nakajima
    • 1
  • Masao Yamashina
    • 1
  • Shinji Nakayama
    • 1
  • Takeo Kusuda
    • 1
  • Yutaku Sakaguchi
    • 1
  • Katsunori Yoshida
    • 1
  • Kazushige Uchida
    • 1
  • Akiyoshi Nishio
    • 1
  • Koichi Matsuzaki
    • 1
  • Kazuichi Okazaki
    • 1
  1. 1.The Third Department of Internal Medicine, Division of Gastroenterology and HepatologyKansai Medical UniversityMoriguchiJapan

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