Project description:We performed RNA-seq, H3K27ac ChIP-seq, and HNF4a ChIP-seq on jejunal intestinal epithelial cells, which are primarily responsible for the absorption of fatty acids, in four conditions: Germ-free (GF), Germ-free plus high fat meal (GF+HFM), ex-GF colonized with a conventional microbiota for 2 weeks (Colonized, CV), and Colonized plus high fat meal (CV+HFM). We, for the first time, map genomewide HFM responsive regulatory regions in the intestine. We identify that in the absence of microbes the HNF4a transcriptional program supports a FAO program in enterocytes while suppressing a proliferation program.
Project description:hydroalcoholic extracts of fecal samples from mice with the following gut-microbiota colonization: conventional, germ-free and oMM12. chromatographic system: 0.1% formic acid and ACN. Mass analyzer QTOF (3D)
Project description:Microbiota-induced cytokine responses participate in gut homeostasis, but the cytokine balance at steady-state and the role of individual bacterial species in setting the balance remain elusive. Using gnotobiotic mouse models, we provide a systematic analysis of the role of microbiota in the induction of cytokine responses in the normal intestine. Colonization by a whole mouse microbiota orchestrated a broad spectrum of pro-inflammatory (Th1, Th17) and regulatory T cell responses. Unexpectedly, most tested complex microbiota and individual bacteria failed to efficiently stimulate intestinal cytokine responses. A potent cytokine-inducing function was however associated with non-culturable host-specific species, the prototype of which was the Clostridia-related Segmented Filamentous Bacterium, and this bacterial species recapitulated the coordinated maturation of T cell responses induced by the whole mouse microbiota. Our study demonstrates the non-redundant role of microbiota members in the regulation of gut immune homeostasis. Germfree (GF) female 8-9-week-old mice were gavaged twice at a 24-hr interval with 0.5 mL of fresh anaerobic cultures of fecal homogenate from SFB mono-associated mice, fresh feces from Cv mice (Cvd) or from a healthy human donor (Hum). All mice were sacrificed on d8, 20 and 60 post-colonization in parallel to age-matched Cv and GF controls. RNA was extracted from ileal tissue, and processed to biotin-labelled cRNA, and then hybridized to the NuGO array (mouse) NuGO_Mm1a520177. Microarray analysis compared gene expression in ileum tissue of all the treatment groups GF, Cv, Cvd, Hum and SFB (N=3 per treatment group per time-point). Data was considered significant when P<0.05 using the Benjamini and Hochberg false discovery method.
Project description:While microbiome and pregnancy are known to alter drug disposition, the interplay of the two physiological factors to impact expression and/or activity of drug processing genes (DPGs) has yet to be elucidated. This study aimed to investigate the effects of microbiome on host hepatic DPGs during pregnancy using conventional (CV) and germ-free (GF) mice. Four groups of female mice were used, namely CV non-pregnant (CVNP), GF non-pregnant (GFNP), CV pregnant (CVP), and GF pregnant (GFP) mice. Pregnant mice examined were on gestation day 15. Transcriptomic and targeted proteomics of hepatic DPGs were profiled using a multi-omics approach. Plasma bile acid and steroid hormone levels were quantified using LC-MS/MS. Cyp3a activities were measured by mouse liver microsome incubations. While the overall trend in pregnancy-induced changes in the expression or activity of hepatic DPGs in CV and GF mice was similar, significant differences in the magnitude of changes were observed. For certain genes, we noticed opposite effects of pregnancy on mRNA and protein expression of DPGs in both CV and GF mice. For instance, the mRNA levels of Cyp3a11, the murine homolog of human CYP3A4, were decreased by 1.7-fold and 3.3-fold by pregnancy in CV and GF mice, respectively. However, the protein levels of Cyp3a11 were increased similarly ~2-fold by pregnancy in both CV and GF mice. Yet, microsome incubations revealed a marked induction of Cyp3a activity by pregnancy that was >5-fold greater in CV mice than that in GF mice. Plasma bile acid and steroid hormone levels were also significantly altered by microbiome and pregnancy, respectively, which may contribute to the differential effects of pregnancy in CV and GF mice. This is the first study to show that microbiome can alter hepatic DPGs in pregnancy.
Project description:IL10 plays a crucial role in regulating immune responses. Although IL10 deficiency induces spontaneous gut inflammation in a commensal-dependent manner, the importance of IL10 signaling in peripheral tissues other than gut remains poorly understood. To determine how peripheral tissues are transcriptionally responded to the commensal-induced IL10 signaling, we performed bulk RNA-seq on the gut, lungs, mesenteric lymph nodes, and kidneys of human fecal microbiota-transplanted (huFMT) IL10 KO, huFMT WT, untreated germ-free (GF) IL10 KO, and untreated GF WT mice.
Project description:This study aimed to quantify and compare the mRNA abundance of all major XPGs in liver and intestine between conventional (CV) and germ-free (GF) mice using RNA-Seq. The CV RNA-Seq data were previously uploaded to GEO database by the same research group (GSE79848). All CV and GF mice were age-matched, and were analyzed under the same conditions (diet, water, bedding, and animal facility).
Project description:Lactobacillus NK2 (L.NK2) is a commensal microbe, isolated from the mouse intestinal feces in our lab. To examine the potential role of L. NK2 in the gut immunity, we monocolonized GF mice with L.NK2. And, we conducted a microarray experiment to compare the transcriptomes of GF and L.NK2-colonized mice intestines under the same experimental condition We used microarrays to detail the global programme of gene expression in intestinal epithelial cells (IEC) and Peyer's patches cells (PP) of GF and L.NK2-colonized mice.
Project description:To investigate the impact of gut microbiota deleting and colonizing on hypothalamic health and function, the tissue samples from the germ-free (GF) pigs and the GF pigs colonized gut microbiota (CG) are used to perform whole RNA-seq for gene expression analysis We then performed long RNAs and small RNAs expression profiling analysis using data obtained from the RNA-seq of 3 different hypothalamus samples from the two groups of pigs