Project description:Commensal microbiota contribute to gut homeostasis and influence gene expression. Intestinal organoid culture closely represent intestinal epithelium and retain intestinal stem cells and dynamic recovery capabilities as well as all major cell types of the intestinal epithelium. We established organoid culture using colon crypts isolated from germ-free (GF), and gnotobiotic mice monocolonized either with the E.coli strain O6K13 (O) or Nissle 1917 strain (N). The expression profiles of these organoids were compared to the organoid culture isolated from conventionally reared (CR) mice in order to disclose genes differentially expressed in response to the change in the intestinal microflora composition.
Project description:This study delineated how small intestinal resident microflora impact gene expression in Paneth cells. Experiment Overall Design: Paneth cells were isolated by laser capture microdissection from the small intestines of germ-free and conventionalized (10 day) mice. RNAs from 3 mice per group were pooled, and duplicate RNAs from each group were amplified and hybridized to Affymetrix arrays.
Project description:This study delineated how small intestinal resident microflora impact gene expression in Paneth cells. Keywords: functional genomics; transcriptional profiling
Project description:This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/ The human-infective whipworm Trichuris trichiura is estimated to infect up to a billion people and is responsible for considerable morbidity, especially in children of developing countries. The closely related species T. muris is a naturally occurring nematode parasite of mice that serves as a remarkably tractable model system for dissecting immune responses and host-parasite relationships. Such studies are of relevance beyond parasitology as helminths have arguably had a significant impact on the evolution of the mammalian immune system. Both Trichuris species reside in the caecum and colon of the host where they burrow their front end for feeding into the intestinal mucosa, thereby breaching the mucus barrier and allowing access of the microflora directly to the epithelium. The interplay of intestinal helminths, the bacterial microflora and the host immune system is currently a research focus in various laboratories (Bancroft et al 2012). This study will study the transcriptional responses of the intestinal mucosa (caecum) from infected and uninfected mice.
Project description:Commensal microbiota contribute to gut homeostasis and influence mucosal gene expression. We harvested mucosal lining of middle and distal part of the small intestine and colon from germ-free (GF) and gnotobiotic mice monocolonized either with the E.coli strain O6K13 (O) or Nissle 1917 strain (N). The expression profiles of the mucosa samples were compared to the corresponding tissue isolated from conventionally reared mice in order to disclose genes differentially expressed in response to the change in the intestinal microflora composition.
Project description:Background. Food can affect the microbial balance in the human intestine, and the ingestion of probiotics may play a role in the current obesity pandemic. The objective of our study was to determine if increased Lactobacillus spp. in the intestinal microflora of mice can promote growth and if changes in the intestinal microflora are associated with modifications in metabolism. Methodology. Female BALBc mice were divided between one control and two experimental groups and inoculated either once or twice with 4×1010 Lactobacillus per animal in PBS or with PBS alone. Fecal samples were collected and tested using qPCR to detect and quantify Lactobacillus spp., Bacteroidetes and Firmicutes. Gene expression by microarray and RT-PCR was studied in liver and adipose tissue. Finally, metabolic parameters in the plasma were tested. Principal Findings. In three independent experiments, we observed an increase in both weight gain and liver weight in mice inoculated with 4×1010 Lactobacillus. Inoculation with Lactobacillus sp. (ostrich) increased the Lactobacillus spp. and Firmicutes DNA copy number in feces. The transcriptional profile of liver tissue from mice inoculated with Lactobacillus sp. (ostrich) was enriched for Gene Ontology terms related to the immune response and metabolic modifications. The mRNA levels of fatty acyl synthase (Fas), sterol regulatory element binding factor 1 (Srebp1c), tumor necrosis factor alpha (Tnf), cytochrome P450 2E1 (Cyp2e1) and 3-phosphoinositide-dependent protein kinase-1 (Pdpk1) were significantly elevated in liver tissue in experimental group animals. In gonadal adipose tissue, the expression of leptin, peroxisome proliferator-activated receptor gamma (Pparg and Srebp1c was significantly higher in experimental group animals, whereas the expression of adiponectin was significantly lower. Conclusions. Alterations in the intestinal microbiota resulted in increased weight gain. Furthermore, increased Lactobacillus spp. in the intestinal microflora of mice inoculated with Lactobacillus sp. (ostrich) resulted in accelerated weight gain, liver enlargement and metabolic changes in the plasma, liver and adipose tissue.