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Research Article

A Crypt-Specific Core Microbiota Resides in the Mouse Colon

Thierry Pédron, Céline Mulet, Catherine Dauga, Lionel Frangeul, Christian Chervaux, Gianfranco Grompone, Philippe J. Sansonetti
David Relman, Editor
Thierry Pédron
Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, Paris, Francea
INSERM U 786, Institut Pasteur, Paris, Franceb
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Céline Mulet
Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, Paris, Francea
INSERM U 786, Institut Pasteur, Paris, Franceb
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Catherine Dauga
Groupe Bioinformatique pour l’Analyse Génomique, Institut Pasteur, Paris, Francec
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Lionel Frangeul
Groupe Bioinformatique pour l’Analyse Génomique, Institut Pasteur, Paris, Francec
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Christian Chervaux
Danone Research, Centre de Recherche Daniel Carasso, Palaiseau, Franced
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Gianfranco Grompone
Danone Research, Centre de Recherche Daniel Carasso, Palaiseau, Franced
Institut Pasteur de Montevideo, Montevideo, Uruguaye
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Philippe J. Sansonetti
Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, Paris, Francea
INSERM U 786, Institut Pasteur, Paris, Franceb
Chaire de Microbiologie et Maladies Infectieuses, Collège de France, Paris, Francef
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David Relman
VA Palo Alto Health Care System
Roles: Editor
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DOI: 10.1128/mBio.00116-12
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  • FIG 1
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    FIG 1

    Bacteria reside in the murine proximal colonic crypts. Warthin-Starry staining (A to D) or FISH (E to H) with the pan-bacterial probe Eub338 at different levels of the intestine of C57BL/6 mice is shown: small intestine (A and E), cecum (B and F), proximal colon (C and G), or distal colon (D and H). Black and white arrows indicate the presence of bacteria. Scale bars, 10 µM.

  • FIG 2
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    FIG 2

    Unweighted Unifrac Bray-Curtis principal coordinate analysis shows dissimilarity in samples. Axis 1, percent variation explained (24.2%); axis 2, percent variation explained (17.17%). Squares and triangles represent crypt and luminal samples, respectively. Each murine strain is represented by a color code (C57BL/6 from provider 1, purple; C57BL/6 from provider 2, blue; BALB/c, green; C3H/HeN, orange).

  • FIG 3
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    FIG 3

    Proportional abundances of the 20 most-represented OTUs in mice belonging to different strains at the crypt and luminal levels. These family-level phylogenetic relative contributions are based on 16S rRNA gene frequencies.

  • FIG 4
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    FIG 4

    Acinetobacter and the Firmicutes are localized mainly in the crypt and lumen of the colon, respectively. Hybridization of an Alexa555-labeled Acinetobacter-specific probe on colonic sections from C57BL/6 (A), BALB/c (B), or C3H/HeN (C) or of an Alexa555-labeled Firmicutes-specific probe on colonic sections from C57BL/6 (D) is shown.

  • FIG 5
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    FIG 5

    Acinetobacter colonizes the colonic crypt. (A) qRT-PCR amplification using phylum- and family-specific primers with DNA extracted from the feces of SPF C57BL/6 mice or from germfree BALB/c mice before and after 7, 15, and 26 days of colonization with conventional microbiota. (B and C) Warthin-Starry (B) or FISH using an Alexa555-labeled Acinetobacter-specific probe (C) on colonic crypt sections after 26 days of colonization of germfree BALB/c mice with a conventional microbiota. Scale bars, 10 µM.

Supplemental Material

  • Figures
  • Additional Files
  • Figure S1

    (A) FISH using the “no-bacteria” probe on different sections of the intestine of C57BL/6 mice. (B) FISH with the pan-bacterial probe Eub338 at the proximal colon and the lumen of germfree C3H/HeN mice. Download Figure S1, PDF file, 1.5 MB.

    Copyright © 2012 Pédron et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Figure S2

    Pie chart showing the percentages of phylotypes (genus level) observed in three murine colonic crypt samples (pilot experiment). Download Figure S2, PDF file, 0.1 MB.

    Copyright © 2012 Pédron et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S1

    Number of reads after multiplex pyrosequencing of murine crypt and luminal samples. Table S1, DOCX file, 0.1 MB.

    Copyright © 2012 Pédron et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Figure S3

    qRT-PCR experiments with DNA extracted from the autoclaved chow using phylum- and family-specific primers. Download Figure S3, PDF file, 0.1 MB.

    Copyright © 2012 Pédron et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S2

    Sequences of primers and probes used in this study. Table S2, DOCX file, 0.1 MB.

    Copyright © 2012 Pédron et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

  • Table S3

    Relative contributions of OTUs at 97% similarity in the different samples resulting from bar-coded pyrosequencing. Table S3, DOCX file, 0.2 MB.

    Copyright © 2012 Pédron et al.

    This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

Additional Files

  • Figures
  • Supplemental Material
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    Supplementary Data

    Files in this Data Supplement:

    • Figure sf01, PDF - Figure sf01, PDF
    • Figure sf02, PDF - Figure sf02, PDF
    • Figure sf03, PDF - Figure sf03, PDF
    • Table st1, DOCX - Table st1, DOCX
    • Table st2, DOCX - Table st2, DOCX
    • Table st3, DOCX - Table st3, DOCX
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A Crypt-Specific Core Microbiota Resides in the Mouse Colon
Thierry Pédron, Céline Mulet, Catherine Dauga, Lionel Frangeul, Christian Chervaux, Gianfranco Grompone, Philippe J. Sansonetti
mBio May 2012, 3 (3) e00116-12; DOI: 10.1128/mBio.00116-12

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A Crypt-Specific Core Microbiota Resides in the Mouse Colon
Thierry Pédron, Céline Mulet, Catherine Dauga, Lionel Frangeul, Christian Chervaux, Gianfranco Grompone, Philippe J. Sansonetti
mBio May 2012, 3 (3) e00116-12; DOI: 10.1128/mBio.00116-12
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