Intest Res.  2015 Jan;13(1):39-49. 10.5217/ir.2015.13.1.39.

Comparative Analysis of Gastrointestinal Microbiota Between Normal and Caudal-Related Homeobox 2 (Cdx2) Transgenic Mice

• Sakamoto H
• Asahara T
• Chonan O
• Yuki N
• Mutoh H
• Hayashi S
• Yamamoto H
• Sugano K

• KMID: 1807375
• DOI: http://doi.org/10.5217/ir.2015.13.1.39

Abstract

BACKGROUND/AIMS
Caudal-related homeobox 2 (Cdx2) is expressed in the human intestinal metaplastic mucosa and induces intestinal metaplastic mucosa in the Cdx2 transgenic mouse stomach. Atrophic gastritis and intestinal metaplasia commonly lead to gastric achlorhydria, which predisposes the stomach to bacterial overgrowth. In the present study, we determined the differences in gut microbiota between normal and Cdx2 transgenic mice, using quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
METHODS
Twelve normal (control) and 12 Cdx2 transgenic mice were sacrificed, and the gastric, jejunal, ileac, cecal and colonic mucosa, and feces were collected. To quantitate bacterial microbiota, we used real-time qRTPCR with 16S rRNA gene-targeted, species-specific primers.
RESULTS
The total numbers of bacteria in the gastric, jejunal, ileac, cecal, and colonic mucosa of the Cdx2 transgenic mice were significantly higher than those of the normal mice. The Bacteroides fragilis group and also Prevotella were not detected in the stomach of the normal mice, although they were detected in the Cdx2 transgenic mice. Moreover, the Clostridium coccoides group, Clostridium leptum subgroup, Bacteroides fragilis group, and Prevotella were not detected in the jejunum or ileum of the normal mice, although they were detected in the Cdx2 transgenic mice. The fecal microbiota of the normal mice was similar to that of the Cdx2 transgenic mice.
CONCLUSIONS
Our results showed the differences in composition of gut microbiota between normal and Cdx2 transgenic mice, which may be caused by the development of gastric achlorhydria and intestinal metaplasia in Cdx2 transgenic mice.

Keyword

Microbiota; Cdx2; Intestinal metaplasia; Gastric achlorhydria; 16S rRNA

MeSH Terms

Achlorhydria
Animals
Bacteria
Bacteroides fragilis
Clostridium
Colon
Feces
Gastritis, Atrophic
Genes, Homeobox*
Humans
Ileum
Jejunum
Metaplasia
Mice
Mice, Transgenic*
Microbiota*
Mucous Membrane
Prevotella
Stomach

Figure

• 

  Fig. 1 Number of gastric bacteria for normal and caudal-related homeobox 2 (Cdx2) transgenic mice. C corresponds to results for Cdx2 transgenic mice, N corresponds to results for normal mice. (T) Number of total bacteria; (1) number of Clostridium coccoides group; (2) number of Clostridium leptum subgroup; (3) number of Bacteroides fragilis group; (6) number of Prevotella ; (8) number of total Lactobacillus ; (9) number of Enterobacteriaceae, (10) number of Enterococcus. The SD value of the bacterial number for each group is shown as a bar. *P<0.05.

• 

  Fig. 2 Number of jejunal bacteria in normal and caudal-related homeobox 2 (Cdx2) transgenic mice. C corresponds to results for Cdx2 transgenic mice, N corresponds to results for normal mice. (T) Number of total bacteria; (1) number of Clostridium coccoides group; (2) number of Clostridium leptum subgroup; (3) number of Bacteroides fragilis group; (4) number of Bifidobacterium; (5) number of Atopobium cluster; (6) number of Prevotella ; (8) number of total Lactobacillus ; (9) number of Enterobacteriaceae ; (10) number of Enterococcus ; (12) number of Staphylococcus. The SD value of the bacterial number for each group is shown as a bar. *P<0.05.

• 

  Fig. 3 Number of ileal bacteria in normal and caudal-related homeobox 2 (Cdx2) transgenic mice. C corresponds to results for Cdx2 transgenic mice, N corresponds to results for normal mice. (T) Number of total bacteria; (1) number of Clostridium coccoides group; (2) number of Clostridium leptum subgroup; (3) number of Bacteroides fragilis group; (4) number of Bifidobacterium; (5) number of Atopobium cluster; (6) number of Prevotella ; (8) number of total Lactobacillus ; (9) number of Enterobacteriaceae ; (10) number of Enterococcus ; (11) number of Streptococcus ; (12) number of Staphylococcus. The SD value of the bacterial number for each group is shown as a bar. *P<0.05.

• 

  Fig. 4 Number of cecal bacteria in normal and caudal-related homeobox 2 (Cdx2) transgenic mice. C corresponds to results for Cdx2 transgenic mice; N corresponds to results for normal mice. (T) Number of total bacteria; (1) number of Clostridium coccoides group ; (2) number of Clostridium leptum subgroup; (3) number of Bacteroides fragilis group; (4) number of Bifidobacterium; (5) number of Atopobium cluster; (6) number of Prevotella ; (8) number of total Lactobacillus ; (9) number of Enterobacteriaceae ; (10) number of Enterococcus ; (11) number of Streptococcus ; (12) number of Staphylococcus. The SD value of the bacterial number for each group is shown as a bar. *P<0.05.

• 

  Fig. 5 Number of colonic bacteria in normal and caudal-related homeobox 2 (Cdx2) transgenic mice. C corresponds to results for Cdx2 transgenic mice; N corresponds to results for normal mice. (T) Number of total bacteria; (1) number of Clostridium coccoides group; (2) number of Clostridium leptum subgroup; (3) number of Bacteroides fragilis group; (4) number of Bifidobacterium; (5) number of Atopobium cluster; (6) number of Prevotella ; (8) number of total Lactobacillus ; (9) number of Enterobacteriaceae ; (10) number of Enterococcus ; (11) number of Streptococcus ; (12) number of Staphylococcus. The SD value of the bacterial number for each group is shown as a bar. *P<0.05.

• 

  Fig. 6 Number of fecal bacteria in normal and caudal-related homeobox 2 (Cdx2) transgenic mice. C corresponds to results for Cdx2 transgenic mice; N corresponds to results for normal mice. (T) Number of total bacteria; (1) number of Clostridium coccoides group; (2) number of Clostridium leptum subgroup; (3) number of Bacteroides fragilis group; (4) number of Bifidobacterium; (5) number of Atopobium cluster; (6) number of Prevotella ; (8) number of total Lactobacillus ; (9) number of Enterobacteriaceae ; (10) number of Enterococcus ; (11) number of Streptococcus ; (12) number of Staphylococcus. The SD value of the bacterial number for each group is shown as a bar. *P<0.05.

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