Chen Ru, Song Jia, Lin Liyuan, Liu Jie, Yang Chaoyong, Wang Wei
Department of Digestive Diseases of Huashan Hospital and Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
Institute of Molecular Medicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Front Mol Biosci. 2021 May 28;8:681938. doi: 10.3389/fmolb.2021.681938. eCollection 2021.
Rat is a widely used mammalian model for gut microbiota research. However, due to the difficulties of individual culture of many of the gut bacteria, much information about the microbial behaviors in the rat gut remains largely unknown. Here, to characterize the growth and division of rat gut bacteria, we apply a chemical strategy that integrates the use of sequential tagging with D-amino acid-based metabolic probes (STAMP) with fluorescence hybridization (FISH) to rat gut microbiota. Following sequential gavages of two different fluorescent D-amino acid probes to rats, the resulting dually labeled gut bacteria provides chronological information of their cell wall synthesis. After taxonomical labeling with FISH probes, most of which are newly designed in this study, we successfully identify the growth patterns of 15 bacterial species, including two that have not been cultured separately in the laboratory. Furthermore, using our labeling protocol, we record cells growing at different growth stages of a complete cell division cycle, which offers a new scope for understanding basic microbial activities in the gut of mammalian hosts.
大鼠是肠道微生物群研究中广泛使用的哺乳动物模型。然而，由于许多肠道细菌的个体培养存在困难，关于大鼠肠道中微生物行为的许多信息在很大程度上仍然未知。在这里，为了表征大鼠肠道细菌的生长和分裂，我们应用了一种化学策略，该策略将基于D-氨基酸的代谢探针的顺序标记（STAMP）与荧光杂交（FISH）相结合，应用于大鼠肠道微生物群。在向大鼠连续灌胃两种不同的荧光D-氨基酸探针后，产生的双重标记肠道细菌提供了它们细胞壁合成的时间信息。在用FISH探针进行分类标记后，其中大部分是本研究新设计的，我们成功地确定了15种细菌的生长模式，包括两种尚未在实验室中单独培养的细菌。此外，使用我们的标记方案，我们记录了在完整细胞分裂周期的不同生长阶段生长的细胞，这为理解哺乳动物宿主肠道中的基本微生物活动提供了新的视角。
