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Ultrastructural and phenotypical diversity of macrophages in the rat ileal mucosa

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Abstract

Several types of macrophages have been reported in the intestinal mucosa, but their histological localization remains ambiguous. Here, we obtained detailed information about ultrastructural and phenotypical diversity of macrophage-like cells (MLCs) in the rat ileal mucosa using immunofluorescent analysis and serial block-face scanning electron microscopy (SBF-SEM). The results revealed that the cells immunopositive for CD68, the pan-macrophage marker, included CD163−CD4+, CD163+CD4+, and CD163−CD4− cells in the lamina propria (LP) of the intestinal villus and around the crypt. CD68+CD4+CD163− cells seemed to be preferentially localized in the intestinal villus, whereas CD68+CD163+CD4+ cells were frequently localized around the crypt. SBF-SEM analysis identified three types of MLCs in the ileal mucosa, which were tentatively named types I–III MLC based on aspects of the 3D-ultrastructure, such as the localization, quantity of lysosomes, endoplasmic reticulum, and exoplasm. Type I and II MLCs were localized in the villous LP, while type III MLCs were localized around the crypt, although type II MLCs were a minor population. All three MLC types extended their cellular processes into the epithelium, with type I MLCs showing the greatest abundance of extended processes. Type I MLCs in the upper portion of the intestinal villus showed a higher level of attachment to intraepithelial lymphocytes (IELs) compared to type III MLCs around the crypt. These findings suggest that macrophages of the rat ileal mucosa differed by region along the longitudinal axis of the villous tip-crypt from the perspective of ultrastructure, cellular composition, localization, and interactions with IELs.

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Acknowledgements

The Ketjen black used to prevent charging of samples was kindly provided by Dr. Ohno.

Funding

This study was supported by the Japan Society for the Promotion of Science (grant numbers: 16K18813 and 20K15902).

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Authors and Affiliations

  1. Laboratory of Histophysiology, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Youhei Mantani, Satoki Nakanishi & Nanami Sakata

  2. Laboratory of Animal Molecular Morphology, Department of Bioresource Science, Graduate School of Agricultural Science, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, Hyogo, 657-8501, Japan

    Toshifumi Yokoyama & Nobuhiko Hoshi

  3. Bio 3D Promotion Group, Application Management Department, JEOL Ltd., 3-1-2 Musashino, Akishima, Tokyo, 196-8558, Japan

    Tomohiro Haruta

  4. Department of Anatomy and Regenerative Biology, Graduate School of Medicine, Osaka City University, 1-4-3, Asahi-machi, Abeno-ku, Osaka, 545-8585, Japan

    Hideto Yuasa

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  1. Youhei Mantani
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  2. Tomohiro Haruta
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  7. Nobuhiko Hoshi
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Correspondence to Youhei Mantani.

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This study was approved by the Institutional Animal Care and Use Committee (permission numbers: 25-06-01 and 30-05-01). All procedures performed in studies involving animals were in accordance with the ethical standards of the institution (the Kobe University Animal Experimentation Regulations) or practice at which the studies were conducted.

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Mantani, Y., Haruta, T., Nakanishi, S. et al. Ultrastructural and phenotypical diversity of macrophages in the rat ileal mucosa. Cell Tissue Res 385, 697–711 (2021). https://doi.org/10.1007/s00441-021-03457-0

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