Specific Localization and Quantification of the Oligo-Mouse-Microbiota (OMM12 ) by Fluorescence In Situ Hybridization (FISH)

Sandrine Brugiroux, David Berry, Diana Ring, Nicolas Barnich, Holger Daims, Bärbel Stecher (Corresponding author)

Publications: Contribution to journalArticlePeer Reviewed

Abstract

The oligo-mouse-microbiota (OMM 12) is a widely used syncom that colonizes gnotobiotic mice in a stable manner. It provides several fundamental functions to its murine host, including colonization resistance against enteric pathogens. Here, we designed and validated specific fluorescence in situ hybridization (FISH) probes to detect and quantify OMM 12 strains on intestinal tissue cross sections. 16S rRNA‒specific probes were designed, and specificity was validated on fixed pure cultures. A hybridization protocol was optimized for sensitive detection of the individual bacterial cells in cryosections. Using this method, we showed that the intestinal mucosal niche of Akkermansia muciniphila can be influenced by global gut microbial community context.

Original languageEnglish
Article numbere548
Number of pages15
JournalCurrent protocols
Volume2
Issue number9
DOIs
Publication statusPublished - 12 Sep 2022

Austrian Fields of Science 2012

  • 106059 Microbiome research
  • 106022 Microbiology

Keywords

  • Animals
  • In Situ Hybridization, Fluorescence/methods
  • Mice
  • Microbiota
  • Oligonucleotide Probes
  • RNA, Ribosomal, 16S/genetics
  • spatial structure
  • intestine
  • microbial ecology
  • oligo-MM
  • altered Schaedler flora
  • sDMDMm2
  • single-cell analysis

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