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Role of RANKL in mouse intestinal epithelial cells

ABSTRACT: Role of RANKL in mouse intestinal epithelial cells

PROVIDER: PRJNA936079 | ENA |

REPOSITORIES: ENA

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Gene expression profile of murine intestinal epithelial cells after RANKL treatment

Project description:RANKL (receptor acrivator of NFkB ligand) is a member of TNF superfamily cytokines. In the gastrointestinal tract, RANKL is expressed in the stromal cells of Peyer's patches, and involved in the development of the specialized intestinal epithelial cells, called M cells. To identify the genes involved in M-cell development, we treated BALB/c mice with recombinant GST-RANKL. After RANKL-treatment, epithelial cells were isolated from small intestine, and used for microarray analysis. Total of 15 samples were analyzed. We generated the Excel sheet for comparing gene expression.

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PeyerM-bM-^@M-^Ys Patch M cells derive from Lgr5+ stem cells require SpiB and are induced by RankL in cultured M-bM-^@M-^XorganoidsM-bM-^@M-^Y

Project description:PeyerM-bM-^@M-^Ys Patches consist of domains of specialized intestinal epithelium overlying Gut-Associated Lymphoid Tissue (GALT). Luminal antigens reach the GALT by translocation through epithelial gatekeeper cells, so-called M cells. We have recently demonstrated that all epithelial cells required for the digestive functions of the intestine are generated from Lgr5-expressing stem cells. Here, we show that M cells also derive from these crypt-based Lgr5 stem cells. The Ets family transcription factor Spi-B, known to control effector functions of bone marrow-derived immune cells, is specifically expressed in M cells. In Spi-B-/- mice, M cells are entirely absent, which occurs in a cell-autonomous fashion. It has been shown that Tnfsf11 (RankL) can induce M cell development in vivo. In intestinal organoid (M-bM-^@M-^Xmini-gutM-bM-^@M-^Y) culture, we show that stimulation with RankL induces SpiB expression within 24hrs and subsequently of other M cell markers. We conclude that RankL-induced expression of Spi-B is essential for Lgr5 stem cell-derived epithelial precursors to develop into M cells. Small intestinal organoids were derived from wildtype (WT) mice. Recombinant mouse RankL (BioLegend) was added to the organoid culture medium in concentrations of 50-200ng/ml and fresh medium was added at day 2 and day 5. At the indicated time points, organoids were harvested for RNA isolation and microarray analysis to look for gene expression changes in reponse to RanKL.

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RNA-seq of mouse small intestinal organoids with an intestinal-epithelial specific deletion of LSD1 compared to wild type

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Mouse Intestinal HMGCR complex

Project description:Mice lacking 3-hydroxy-3-methylglutaryl-coenzyme A reductase (Hmgcr) in intestinal villus and crypt epithelial cells were generated using a Villin-Cre transgene. Label free proteome profiling was measure for Wild type and KO mouse.

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