Institute of Molecular Medicine (IMM), Shanghai Key Laboratory for Nucleic Acid Chemistry and Nanomedicine, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China.
The MOE Key Laboratory of Spectrochemical Analysis and Instrumentation, Key Laboratory for Chemical Biology of Fujian Province State Key Laboratory of Physical Chemistry of Solid Surfaces, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China.
Angew Chem Int Ed Engl. 2020 Jul 13;59(29):11923-11926. doi: 10.1002/anie.202004703. Epub 2020 May 14.
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.
在此，我们提出了一种基于代谢 D-氨基酸的标记和原位杂交辅助（MeDabLISH）的策略，用于定量分析肠道细菌的固有代谢状态。我们发现，细菌的荧光 D-氨基酸（FDAA）标记强度与它们的时间和稳态代谢状态高度相关。然后，在对体内 FDAA 标记的小鼠肠道微生物群中的细菌属进行分类鉴定后，通过相应的荧光原位杂交（FISH）探针，使用 FISH 信号来区分属，用 FDAA 信号来指示它们的基础代谢水平，通过流式细胞术对不同肠道细菌属的代谢活性进行定量。结果发现，小鼠微生物群中的革兰氏阴性菌在白天的代谢活性更强，而革兰氏阳性菌在夜间的活性更高。我们的策略将有助于深入了解高度复杂的微生物群。
