title: Nature of bacterial colonization influences transcription of mucin genes in mice during the first week of life.
abstract: BACKGROUND
  Postnatal regulation of the small intestinal mucus layer is potentially important in the development of adult gut functionality. We hypothesized that the nature of bacterial colonization affects mucus gene regulation in early life.We thus analyzed the influence of the presence of a conventional microbiota as well as two selected monocolonizing bacterial strains on the transcription of murine genes involved in mucus layer development during the first week of life.Mouse pups (N = 8/group) from differently colonized dams: Germ-free (GF), conventional specific pathogen free (SPF), monocolonized with either Lactobacillus acidophilus NCFM (Lb) or Escherichia coli Nissle (Ec) were analyzed by qPCR on isolated ileal tissue sections from postnatal days 1 and 6 (PND1, PND6) after birth with respect to: (i) transcription of specific genes involved in mucus production (Muc1-4, Tff3) and (ii) amounts of 16S rRNA of Lactobacillus and E. coli. Quantification of 16S rRNA genes was performed to obtain a measure for amounts of colonized bacteria.
  
  
  RESULTS
  We found a microbiota-independent transcriptional increase of all five mucus genes from PND1 to PND6. Furthermore, the relative level of transcription of certain mucus genes on PND1 was increased by the presence of bacteria. This was observed for Tff3 in the SPF, Ec, and Lb groups; for Muc2 in SPF; and for Muc3 and Muc4 in Ec and Lb, respectively.Detection of bacterial 16S rRNA genes levels above the qPCR detection level occurred only on PND6 and only for some of the colonized animals. On PND6, we found significantly lower levels of Muc1, Muc2 and Muc4 gene transcription for Lb animals with detectable Lactobacillus levels as compared to animals with Lactobacillus levels below the detection limit.
  
  
  CONCLUSIONS
  In summary, our data show that development of the expression of genes encoding secreted (Muc2/Tff3) and membrane-bound (Muc1/Muc3/Muc4) mucus regulatory proteins, respectively, is distinct and that the onset of this development may be accelerated by specific groups of bacteria present or absent at the mucosal site.
authors.affiliation: Gut Ecology Group, Department of Food Microbiology, National Food Institute, Technical University of Denmark, Søborg, Denmark. adbe@food.dtu.dk
authors.forename: Anders
authors.initials: A
authors.lastname: Bergström
authors.forename: Matilde B
authors.initials: MB
authors.lastname: Kristensen
authors.forename: Martin I
authors.initials: MI
authors.lastname: Bahl
authors.forename: Stine B
authors.initials: SB
authors.lastname: Metzdorff
authors.forename: Lisbeth N
authors.initials: LN
authors.lastname: Fink
authors.forename: Hanne
authors.initials: H
authors.lastname: Frøkiaer
authors.forename: Tine R
authors.initials: TR
authors.lastname: Licht
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D004926:Escherichia coli / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D005856:Germ-Free Life / Q000235:genetics; D007421:Intestine, Small / Q000378:metabolism* / Q000382:microbiology; D007779:Lactobacillus acidophilus / Q000502:physiology*; D054892:Metagenome / Q000502:physiology*; D051379:Mice; D059013:Microbial Consortia / Q000235:genetics; D018396:Mucin-1 / Q000235:genetics / Q000378:metabolism; D055262:Mucin-2 / Q000235:genetics / Q000378:metabolism; D055263:Mucin-3 / Q000235:genetics / Q000378:metabolism; D055264:Mucin-4 / Q000235:genetics / Q000378:metabolism; D009077:Mucins / Q000235:genetics / Q000378:metabolism; D009093:Mucus / Q000378:metabolism* / Q000382:microbiology; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014158:Transcription, Genetic; D000071165:Trefoil Factor-3
chemical_list: C524626:Muc2 protein, mouse; C524641:Muc3 protein, mouse; C524659:Muc4 protein, mouse; D018396:Mucin-1; D055262:Mucin-2; D055263:Mucin-3; D055264:Mucin-4; D009077:Mucins; D012336:RNA, Ribosomal, 16S; C493263:Tff3 protein, mouse; D000071165:Trefoil Factor-3; C577056:muc1 protein, mouse