Enterocyte proliferation and apoptosis in the caudal small intestine is influenced by the composition of colonizing commensal bacteria in the neonatal gnotobiotic pig.
We previously reported marked differences in small intestinal morphology, including changes in crypt depth and villous height, after inoculation of germ-free pigs with different bacterial species. In an attempt to identify the mechanisms governing changes in villous morphology associated with bacterial colonization, 2 gnotobiotic experiments were performed. In each experiment, 16 piglets were allocated to 4 treatment groups including germ-free (GF), monoassociation with Lactobacillus fermentum (LF) or Escherichia coli (EC), or conventionalized with sow feces (SF). Piglets were reared under gnotobiotic conditions until 14 d of age, at which time whole intestinal tissue and enterocytes were collected for histological, gene expression, and protein analysis. Proliferating cell nuclear antigen, tumor necrosis factor alpha (TNFalpha), Fas ligand (FasL), CD3epsilon, caspase 3 (casp3), and toll-like receptors (TLR)2, 4, and 9 expression were measured by quantitative PCR. Activated casp3 was measured by Western blot. Increased abundance of activated casp3 and transcripts encoding proliferating cell nuclear antigen, TNFalpha, CD3epsilon, and FasL was observed in SF and EC treatment groups compared with GF and LF. Expression of TLR2 was increased (P < 0.05) in the SF treatment and tended to be greater (P < 0.08) in EC relative to LF and GF. Results indicate that conventional bacteria and E. coli but not L. fermentum increase overall cell turnover by stimulating increased apoptosis through the expression of FasL and TNFalpha and by increasing cell proliferation. The differential regulation of TLR expression indicates that microbially induced changes may be mediated in part by these receptors. Induction of inflammatory responses and activation of apoptosis through death receptors appears to play a significant role in enterocyte turnover mediated by commensal bacteria.
Willing
B P
BP
Department of Animal and Poultry Science, University of Saskatchewan, 51 Campus Drive, Saskatoon, Saskatchewan, Canada.
Van Kessel
A G
AG
D000818:Animals; D017209:Apoptosis; D020169:Caspases / Q000032:analysis / Q000378:metabolism; D002455:Cell Division; D004912:Erythrocytes / Q000502:physiology*; D004926:Escherichia coli / Q000254:growth & development; D053222:Fas Ligand Protein / Q000032:analysis / Q000378:metabolism; D005243:Feces / Q000382:microbiology; D005856:Germ-Free Life; D007421:Intestine, Small / Q000276:immunology* / Q000382:microbiology* / Q000473:pathology; D052197:Limosilactobacillus fermentum / Q000254:growth & development; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D011897:Random Allocation; D015398:Signal Transduction; D013552:Swine; D051193:Toll-Like Receptors / Q000032:analysis / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000378:metabolism