Simultaneous real-time analysis of Paneth cell and intestinal stem cell response to interferon-γ by a novel stem cell niche tracking method

Yuki Yokoi; Takahiro Adachi; Rina Sugimoto; Mani Kikuchi; Tokiyoshi Ayabe; Kiminori Nakamura

doi:10.1016/j.bbrc.2021.01.050

Simultaneous real-time analysis of Paneth cell and intestinal stem cell response to interferon-γ by a novel stem cell niche tracking method

Biochem Biophys Res Commun. 2021 Mar 19:545:14-19. doi: 10.1016/j.bbrc.2021.01.050. Epub 2021 Jan 30.

Authors

Yuki Yokoi¹, Takahiro Adachi², Rina Sugimoto³, Mani Kikuchi¹, Tokiyoshi Ayabe⁴, Kiminori Nakamura⁵

Affiliations

¹ Innate Immunity Laboratory, Department of Cell Biological Science, Faculty of Advanced Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan.
² Department of Immunology, Medical Research Institute, Tokyo Medical and Dental University, 1-5-45 Yushima, Bunkyo-ku, Tokyo, 113-8510, Japan.
³ Innate Immunity Laboratory, Graduate School of Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan.
⁴ Innate Immunity Laboratory, Department of Cell Biological Science, Faculty of Advanced Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan; Innate Immunity Laboratory, Graduate School of Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan.
⁵ Innate Immunity Laboratory, Department of Cell Biological Science, Faculty of Advanced Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan; Innate Immunity Laboratory, Graduate School of Life Science, Hokkaido University, Kita-21, Nishi-11, Kita-ku, Sapporo, Hokkaido, 001-0021, Japan. Electronic address: kiminori@sci.hokudai.ac.jp.

PMID: 33529805
DOI: 10.1016/j.bbrc.2021.01.050

Abstract

Paneth cells and Lgr5+ intestinal stem cells (Lgr5+ ISCs) constitute the stem cell niche and maintain small intestinal epithelial integrity by recognizing various niche factors derived from subepithelial cells and external antigens. Although it has been known that interferon-γ (IFN-γ), a Th1 cytokine, is associated with intestinal epithelial disruption during inflammation as a niche factor, dynamics of Paneth cells and Lgr5+ ISCs in response to IFN-γ remain to be understood. Here we show that CAG-tdTomato;Lgr5-EGFP (CT-LE) mice generated in this study enable to identify Paneth cells and Lgr5+ ISCs separately by fluorescence signals. Lgr5+ ISCs underwent cell death a little earlier than Paneth cells in response to IFN-γ by simultaneous tracking using CT-LE mice. In addition, the timing of cell death in most Paneth cells overlapped with Lgr5+ ISCs, suggesting that Paneth cell depletion is induced directly by IFN-γ. Taken together, we established a novel simultaneous stem cell niche tracking method and clarified the involvement of both Paneth cells and Lgr5+ ISCs in stem cell niche damage induced by IFN-γ, further contribute to understanding the mechanism for maintaining intestinal homeostasis by stem cell niche.

Keywords: Interferon-γ; Intestinal stem cell; Live imaging; Paneth cell; Stem cell niche.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Death / drug effects
Cell Death / physiology
Computer Systems
Homeostasis / drug effects
Homeostasis / physiology
Interferon gamma Receptor
Interferon-gamma / pharmacology*
Interferon-gamma / physiology
Intestinal Mucosa / drug effects*
Intestinal Mucosa / pathology*
Intestinal Mucosa / physiology
Mice
Mice, Transgenic
Paneth Cells / drug effects*
Paneth Cells / pathology*
Paneth Cells / physiology
Receptors, G-Protein-Coupled / metabolism
Receptors, Interferon / metabolism
Stem Cell Niche / drug effects
Stem Cell Niche / physiology
Stem Cells / drug effects*
Stem Cells / pathology*
Stem Cells / physiology

Substances

Lgr5 protein, mouse
Receptors, G-Protein-Coupled
Receptors, Interferon
Interferon-gamma