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High fat diet decrease diversity of extracellular DNA of mice gut microbiota
Author(s) -
Sun Jin,
Li Ya,
Wang Panpan,
Shi Yonghui
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.lb204
Subject(s) - mucus , biology , bacteria , microbiology and biotechnology , genetics , ecology
Different CpG DAN motif from gut bacteria shows stimulatory or suppressive effect on host immune cells or epithelial cell. Bacterial extracellular DNA (eDNA) in mucus have greater potential to penetrate mucus layer and to be sensed by host immune cell. To investigated the influence of high fat diet on the composition of these eDNA. Mucus eDNA was prepared from control and high fat diet treated C57BL/6 mice. Their diversities were analyzed by T‐RFLP. Mucus layer eDNA were observed in situ using laser scanning confocal fluorescence microscope after TOTO strain on tissue section. The result revealed that eDNA were enriched in mucus layer, some eDNA even locate in. Mucus could be prepared using PBS containing 0.5 M dithiothreitol. Than eDNA were obtained using two step phenol extraction. For eDNA specific T‐RFLP, 16s rRNA gene should be amplified using 334F/939R primer, the PCR products should be treated using restriction endonuclease of AluI and DdeI. Principal component analysis of T‐RFLP results revealed that eDNA was different intracellular DNA. In small intestine, major eDNA were from bacteria Bacaeroides phylum revealed by quantitative PCR analysis. High fat diet significantly decreased diversity of mucus eDNA. Support or Funding Information This work was supported by National Natural Science Foundation of China (No. 31201805) 1Mucus layer (left) and extracellular DNA (right) of mouse colon.2Principal component analyses of forward and reverse TRFs based on its size and relative weight