A new approach for analyzing an adhesive bacterial protein in the mouse gastrointestinal tract using optical tissue clearing
A new approach for analyzing an adhesive bacterial protein in the mouse gastrointestinal tract using optical tissue clearing
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DOI：
10.1038/s41598-019-41151-y
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发表时间：
2019-03-18
影响因子：
4.6
通讯作者：
Okada, Nobuhiko
中科院分区：
文献类型：
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作者：
Nishiyama, Keita;Sugiyama, Makoto;Okada, Nobuhiko
几种细菌兼职蛋白充当粘附因子，其对于胃肠道（GI）的细菌定殖是重要的。然而，关于兼职蛋白在胃肠道中的粘附特性知之甚少。在这里，我们描述了一种新的方法，通过使用组织光学透明化方法，以延长因子Tu（EF-Tu）为例，在整个胃肠道中可视化兼职蛋白质包被的荧光微珠的定位。作为细菌细胞表面定位的蛋白质模拟物，来自罗伊氏乳杆菌的重组EF-Tu固定在微珠上。EF-Tu涂层促进了微珠与Caco-2细胞单层的相互作用。接下来，将微珠口服给药于小鼠。在浓度递增的果糖水溶液中清除GI全组织。在给药后1小时，微珠从胃扩散到盲肠，3小时后，它们扩散到整个肠道。在下消化道中，EF-Tu珠比未涂覆的对照珠显著更丰富，表明EF-Tu在微珠在胃肠道中的持久性中起重要作用。新方法将有助于评估兼职蛋白质如何介导细菌定植。
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.