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Quantification and localization of erythropoietin-receptor-expressing cells in the liver of Xenopus laevis

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Abstract

Erythropoiesis occurs in the African clawed frog, Xenopus laevis and is mediated by erythropoietin (xlEPO), a primary regulator of this process. Previously, we have shown that the xlEPO receptor (xlEPOR), which is expressed by erythroid progenitors that respond to xlEPO, is found predominantly in the liver. The aim of the present study was to determine the dynamics of erythropoiesis in the livers of normal and anemic X. laevis by identifying the number and precise location of mature and immature erythrocytes. We quantified mature and immature erythrocyte numbers by o-dianisidine staining or immunohistochemistry and investigated the dynamics of erythropoiesis in normal, acute hemolytic and blood-loss states by in vivo cell proliferation assays with 5-bromo-2′-deoxyuridine (BrdU). We detected 0.12×108 xlEPOR+ BrdU+ cells in the liver of the normal X. laevis at 24 h after BrdU injection. Frogs presenting with acute hemolytic anemia and pancytopenia show a 10-fold increase in the number of xlEPOR+/BrdU+ cells (approximately 1.30×108 cells) in the liver. The xlEPOR+ cells are found predominantly on the inner wall of hepatic sinusoids. Hematopoietic progenitors that undergo slow cell cycling were also observed in the hepatic sinusoids. This study clarifies the rate of production of mature and immature erythrocytes per day in the liver of X. laevis and the way that these cell numbers change in response to anemia.

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Authors and Affiliations

  1. Department of Integrative Bioscience and Biomedical Engineering, Graduate School of Advanced Science and Engineering, Center for Advanced Life and Medical Science, Waseda University, TWIns Building, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan

    Takehito Okui, Yusuke Yamamoto, Kazumichi Nagasawa, Yuka Yonezuka & Takashi Kato

  2. Department of Biology, School of Education, Waseda University, TWIns Building, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan

    Shun Maekawa, Youichi Aizawa & Takashi Kato

Authors
  1. Takehito Okui
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  2. Yusuke Yamamoto
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  3. Shun Maekawa
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  4. Kazumichi Nagasawa
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  5. Yuka Yonezuka
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  6. Youichi Aizawa
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  7. Takashi Kato
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Corresponding author

Correspondence to Takashi Kato.

Additional information

This work was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (Tokyo, Japan) and by Waseda University (Tokyo, Japan) grants for special research projects. Part of this study was performed as a component of a Private University “High-Tech Research Center” project supported by the Japanese Ministry of Education, Culture, Sports, Science, and Technology. A part of this study was performed as a MEXT-Supported Program for the Strategic Research Foundation at Private Universities.

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Okui, T., Yamamoto, Y., Maekawa, S. et al. Quantification and localization of erythropoietin-receptor-expressing cells in the liver of Xenopus laevis . Cell Tissue Res 353, 153–164 (2013). https://doi.org/10.1007/s00441-013-1624-8

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  • DOI: https://doi.org/10.1007/s00441-013-1624-8

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