Influence of the intrinsic gut microbiota on transcriptional regulation of genes involved in the early life development of intestinal epithelial integrity

Anders Bergström, Matilde Bylov Kristensen, Hanne Frøkjær, Tine Rask Licht

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Abstract

The interplay between the gut microbiota and the integrity of the intestinal mucus layer is important both in the maintenance of the epithelial barrier as part of the innate immune defense, and in the conservation of gut homeostasis. Interesting parameters are the mucins, which protect the mucosal surfaces of all epithelial linings by physical or specific hindrance of pathogenic species e.g. virus and bacteria. Moreover, the proteins constituting the tight junctions in the apical membrane of the epithelial cells are important as they take part in controlling, which substances can penetrate the barrier from the gut lumen to the blood circulation. Previously, it has been shown that the early life mucus layer in germ-free mice has a distinctly different composition than in conventionally colonized animals. In this study, four groups of differently colonized mice were used to analyze mRNA expression by real-time quantitative PCR of relevant mucin (Muc1-4) and tight junction genes (JAM-A, E-Cad, Tjp-1) on RNA purified from isolated ileum samples (n=8 in each group). The groups were: 1) Germ Free (GF), 2) Specific Pathogen Free (SPF) i.e. – “conventional microbiota”, 3) NCFM (GF monocolonized with Lactobacillus NCFM), 4) E.coli (GF monocolonized with E.coli). Ileal samples were taken on day 1 and day 6 after birth in order to analyze early life developmental parameters. On the day 6 samples, mucin-related mRNA’s showed significantly higher expression levels in the GF animals compared to the SPF animals, possibly as part of protective mechanism. Monocolonization with Lactobacillus NCFM and E.coli seemed to decrease levels towards levels observed in the SPF animals (except for Muc-3 in E.coli). Two of the tight junction genes (JAM-A, E-Cad) showed similar tendencies, whereas Tjp-1 showed high levels in both GF and SPF. Comelli EM et al (2008) have shown very similar results on the mucin genes, when colonizing with human adult or baby “full” microbiota. This is the first study with monocolonization however. Finally, we observed inverse correlation between Muc-1 and Lactobacillus 16S rRNA expression. The analysis of the day 1 samples is ongoing and results will be presented at the meeting
Original languageEnglish
Title of host publication2nd TNO Beneficial Microbes Conference : Abstracts of lectures and posters
Number of pages61
Publication date2010
Publication statusPublished - 2010
Event2nd TNO Beneficial Microbes Conference: International conference on the health impact and future potential of beneficial microbes - NH Leeuwenhorst Conference Centre, Noordwijkerhout, Netherlands
Duration: 15 Mar 201017 Mar 2010
Conference number: 2

Conference

Conference2nd TNO Beneficial Microbes Conference
Number2
LocationNH Leeuwenhorst Conference Centre
CountryNetherlands
CityNoordwijkerhout
Period15/03/201017/03/2010

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