Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Ileum mucosa transcriptome profiling of two laying hen strains at five time points spanning egg production phases during 10 to 60 weeks of age

ABSTRACT: In this study, we deep-sequenced the transcriptomic profile of the ileum mucosa in two laying hen strains (Lohmann Brown-Classic and Lohmann LSL-Classic) at 10, 16, 24, 30, and 60 weeks of age. These five time points span the initial, peak and end of the lay periods in which the hens critically experience physiological changes and need optimal diets to meet both growth and egg production requirements. Therefore, a comparative transcriptome profiling of the ileum mucosa may lead to a better understanding of molecular mechanisms underlying genetics, age and nutrition factors of layers.

INSTRUMENT(S): Illumina HiSeq 2500

ORGANISM(S): Gallus gallus

SUBMITTER: Nares Trakooljul

PROVIDER: E-MTAB-9188 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Gut dysbiosis and host genetics are implicated as causative factors in inflammatory bowel disease, yet mechanistic insights are lacking. Longitudinal analysis of ulcerative colitis patients following total colectomy with ileal anal anastomosis (IPAA) where >50% develop pouchitis, offers a unique setting to examine cause vs. effect. To recapitulate human IPAA, we employed a mouse model of surgically-created blind self-filling (SFL) and self-emptying (SEL) ileal loops. SFL exhibit fecal stasis due to directional peristalsis motility oriented towards away from the loop end, whereas SEL remain empty. In wild type mice, SFL, but not SEL, develop pouch-like microbial communities without accompanying active inflammation. However, in genetically susceptible IL-10-/- deficient mice, SFL, but not SEL, exhibit severe inflammation and mucosal transcriptomes resembling human pouchitis. Germ-free IL10-/- mice conventionalized with wild type SFL, but not SEL, microbiota, develop severe colitis. These data demonstrate an essential role for fecal stasis, gut dysbiosis, and genetic susceptibility and offer insights into human pouchitis and ulcerative colitis. All animal protocols were approved by IACUC at the University of Chicago. All animals were C57Bl/6 mice that were bred and housed under standard 12:12 light/dark conditions at the University of Chicago. Female mice aged 6-8 weeks were fed ad libitum gel diet 76A (Cat# 72-07-5022, Clear H20, Portland, ME) for 5-days prior to surgery to prevent obstruction at the anastomosis. Animals were anesthetized with ketamine/xylazine. Aseptic surgery was performed to resect 2.5cm of ileum 3cm proximal to the ileal-cecal value with anastomosis to the ileum using 8-0 suture (Figure 1a). The abdominal wall was closed with interrupted 4-0 silk suture and skin was closed with staples. Analgesics (betanorphine mg/kg BW) were provided post-operatively. After 5 weeks, mice were humanely euthanized. Intestinal loops were collected for RNA, protein, and histology. Loop, sham ileum, and sham colon contents were collected and snap frozen at −80°C for microbiota analysis. Human biopsies and stool samples were obtained under IRB approval and privacy protocols were followed. Our initial work demonstrated up regulation of TLR4 signaling in the mucosa of self-filling ileal loops. We hypothesized that TLR4 may be in-part responsible for mediating the metaplasia and inflammatory responses observed. Therefore, TLR4 KO mice were used to test this hypothesis and subsequently demonstrated attenuated responses in these parameters.

Dataset Information

Ileum mucosa transcriptome profiling of two laying hen strains at five time points spanning egg production phases during 10 to 60 weeks of age

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