Project description:Using Affymetrix data analysis, important signalling pathways and transcription factors relevant to gut inflammation and anti-inflammatory action of probiotics were identified using the clinically validated probiotic VSL#3 and the IL10-knockout mouse, an animal model for inflammatory bowel disease. VSL#3 increased expression of genes in volved in PPAR signalling and metabolism of xenobiotics and decreased expression of genes involved in immune response/inflammatory response. IL10-knockout (IL10-KO) and wildtype (WT) mice housed under specific pathogen free (SPF) conditions were sacrificed at 24 weeks by cervical dislocation. The study is comprised of two independent Microarray experiments. Microarray experiment1 compares gene expression of IL10-KO and WT colon tissue. For microarray analysis RNA was extracted from the colon tissue of each mouse (WT n=7, IL10-KO n=6). Microarray experiment2 compares gene expression of WT and IL10-KO mice fed with either placebo or probiotic VSL#3. IL10-KO and WT mice were fed with placebo or 1.3x109 cfu of lyophilized VSL#3 bacteria post weaning for 21 weeks. For microarray analysis RNA was extracted from the caecum tissue of each mouse (WT Placebo n=6, IL10-KO Placebo n=6, IL10-KO VSL#3 n=6).
Project description:We used 16S V3/V4 region amplification to evaluate the composition of bacteria species in mouse fecal pellets. Fecel pellets were collected from young-adult (12 weeks old) wild type C57Bl/6 mice and aged (72 weeks old) wild type C57Bl/6 mice after 21 days of vehicle or antibiotics treatment (to induce gut microbiota depletion). In one sequencing round, we sequenced a total of 12 different fecal samples (3 young control, 3 aged control, 3 young depleted gut microbiota (ABX) and 3 aged depleted gut microbiota (ABX)). Amplicons were indexed using the Nextera XT Index Kit and pooled into a library for Illumina sequencing.