A new approach for analyzing an adhesive bacterial protein in the mouse gastrointestinal tract using optical tissue clearing

Keita Nishiyama, Makoto Sugiyama, Hiroki Yamada, Kyoko Makino, Sayaka Ishihara, Takashi Takaki, Takao Mukai, Nobuhiko Okada

研究成果: Article査読

6 被引用数 (Scopus)

抄録

Several bacterial moonlighting proteins act as adhesion factors, which are important for bacterial colonization of the gastrointestinal (GI) tract. However, little is known about the adherence properties of moonlighting proteins in the GI tract. Here, we describe a new approach for visualizing the localization of moonlighting protein-coated fluorescent microbeads in the whole GI tract by using a tissue optical clearing method, using elongation factor Tu (EF-Tu) as an example. As a bacterial cell surface-localized protein mimic, recombinant EF-Tu from Lactobacillus reuteri was immobilized on microbeads. EF-Tu-coating promoted the interaction of the microbeads with a Caco-2 cell monolayer. Next, the microbeads were orally administered to mice. GI whole tissues were cleared in aqueous fructose solutions of increasing concentrations. At 1 h after administration, the microbeads were diffused from the stomach up to the cecum, and after 3 h, they were diffused throughout the intestinal tract. In the lower digestive tract, EF-Tu-beads were significantly more abundant than non-coated control beads, suggesting that EF-Tu plays an important role in the persistence of the microbeads in the GI tract. The new approach will help in evaluating how moonlighting proteins mediate bacterial colonization.

本文言語English
論文番号4731
ジャーナルScientific reports
9
1
DOI
出版ステータスPublished - 2019 12 1
外部発表はい

ASJC Scopus subject areas

  • 一般

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