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Quantification of Bacterial Metabolic Activities in the Gut by d ‐Amino Acid‐Based In Vivo Labeling
Author(s) -
Lin Liyuan,
Song Jia,
Du Yahui,
Wu Qiuyue,
Gao Juan,
Song Yanling,
Yang Chaoyong,
Wang Wei
Publication year - 2020
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.202004703
Subject(s) - gut flora , bacteria , biology , amino acid , in vivo , biochemistry , in situ , fluorescence in situ hybridization , flow cytometry , metabolic pathway , metabolism , microbiology and biotechnology , chemistry , genetics , gene , organic chemistry , chromosome
Herein, we propose a metabolic d ‐amino acid‐based labeling and in situ hybridization‐facilitated (MeDabLISH) strategy for the quantitative analysis of the indigenous metabolic status of gut bacteria. The fluorescent d ‐amino acid (FDAA)‐based labeling intensities of bacteria were found to highly correlate with their temporal and steady‐state metabolic status. Then, after taxonomic identification of bacterial genera in the in vivo FDAA‐labeled mouse gut microbiota, by corresponding fluorescence in situ hybridization (FISH) probes, the metabolic activities of different gut bacterial genera are quantified by flow cytometry, using FISH signals to differentiate genera and FDAA signals to indicate their basal metabolic levels. It was found that Gram‐negative genera in the mouse microbiota have stronger metabolic activities during the daytime, and Gram‐positive genera have higher activities at the night. Our strategy will be instrumental in deepening our understanding of the highly complex microbiota.