A profile of murine gastric epithelial cells: Parietal, Zymogenic, Pit
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE5018 | GEO | 2006/10/20
SECONDARY ACCESSION(S): PRJNA95805
REPOSITORIES: GEO
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Project description:Continuous regeneration of digestive enzyme (zymogen) secreting chief cells is a normal aspect of stomach function that is disrupted in pre-cancerous lesions. Regulation of zymogenic cell (ZC) differentiation is poorly understood. Here we profile Parietal, Pit, and Zymogenic cells for comparison and study. Experiment Overall Design: Samples were obtained through Laser-capture microdisection of gastric epithelium. The three samples come from enrichments for Zymogenic Cells (ZC), Parietal Cells (PC), and Mucus Pit Cells (Pit). Experiment Overall Design: Individual parietal cells (visualized by DBA-positivity and autofluorescence) from well-oriented gastric units were dissected (PixCell II LCM apparatus, 7.5-µm spot diameter; CapSure HS LCM caps, Arcturus, Mountain View, CA) one-at-a-time from the pit zone and collected for GeneChips. Pit cells (E-cadherin-positive, DBA-negative) were then collected from the same gastric units. Only the pit cells in a 2-3-cell-thick region at the apex of the gastric unit â?? but not yet upon the gastric surface â?? were taken. ZCs (E-cadherin-positive, GSII/DBA-negative cells in the base zones) were collected from different slides in corpus gastric units after potentially contaminating basal parietal cells had first been dissected and discarded. 3000-5000 individual cells from each cell lineage were isolated from 4-5 individual mice, and RNA was purified by PicoPure kit (Arcturus). RNA integrity was verified, and RNAs for each lineage were pooled from multiple mice and multiple dissections to make 20 ng total RNA, which was then amplified, labeled, and fragmented (by the Arcturus RiboAmp HS kit followed by the RNA Amplification and Labeling Kit from Enzo Life Sciences). The resulting biotinylated cRNA probes were hybridized to Affymetrix (Santa Clara, CA) GeneChip® Mouse Genome 430 2.0 microarrays.
2008-06-13 | E-GEOD-5018 | biostudies-arrayexpress
Project description:The transcription factor MIST1 is required for final maturation of secretory cells of diverse tissues, including gastric digestive-enzyme secreting zymogenic (chief) cells (ZCs). Here, we show that MIST1 directly activates RAB26, RAB3D and several other genes.
2009-12-17 | GSE16924 | GEO
Project description:MIST1 is a bHLH transcription factor that is necessary for the maturation of gastric zymogenic cells as they differentiate from their precursor mucous neck cells. In this experiment, mucous neck cells and zymogenic cells of normal, adult C57BL/6 and MIST1 knockout mice were laser-capture microdissected in order to determine MIST1-dependent, zymogenic cell specific gene expression.
2013-01-12 | GSE43441 | GEO
Project description:In this experiment, mucous neck cells from the gastric epithelium of normal, adult C57/B6 mice were laser-capture microdissected to determine gene expression in neck cells relative to pit cells, parietal cells, and zymogenic cells, whose expression profiles were previously deposited in GEO.
2013-05-31 | GSE42946 | GEO
Project description:The transcription factor MIST1 is required for final maturation of secretory cells of diverse tissues, including gastric digestive-enzyme secreting zymogenic (chief) cells (ZCs). Here, we show that MIST1 directly activates RAB26, RAB3D and several other genes. Experiment Overall Design: We used microarrays to determine genes upregulated following transient MIST1 transfection. Two gastric cell lines were used: HGC-27 and AGS. Three chips were generated from each cell line: parental (untransfected), GFP+empty vector (a control for effects of transfection), and GFP+MIST1. The latter two chips were generated from cells flow sorted based on GFP fluorescence to isolate cells with high levels of transfection. Chips were analyzed by dChip to identify genes expressed in both MIST1-transfected cell populations relative to their respective controls in multiple Boolean 'AND' comparisons.
2010-01-30 | E-GEOD-16924 | biostudies-arrayexpress
Project description:Mist1+ cells and parietal cells in mouse stomach were separatedly sorted, and RNAs were isolated. Mist1 (also known as Bhlha15) is expressed in gastric chief cells and gastric stem cells in mice. However, more specific genes for each population needs to be identified to better understand the precise biology in these cell populations. In order to address cell specific gene signature, we separately sorted Mist1+ gastric chief cells and Mist1+ gastric stem cells by FACS, and performed microarray analysis. Mist1+ gastric chief cells were sorted by using Mist1-CreERT; R26-TdTomato mouse stomach, immediately after tamoxifen administration. Mist1+ gastric stem cells were sorted by chief cell-ablated Mist1-CreERT; R26-TdTomato mouse stomach, combining with Lgr5-DTR mice. Lgr5-expressing chief cells were ablated by giving DT into these mice. As a control, acid-secreting gastric parietal cell samples were used. Mice were treated with or without Lgr5-DT ablation before sorting.
2019-06-25 | GSE133205 | GEO
Project description:Mice were infected with Helicobacter pylori strain SS1 for 2, 7, 14 or 28 days or left uninfected. They were sacrificed at the appropriate times, the stomachs were removed and one half was plated for colony counts. The other half was embedded in OCT and cryosections were produced for 6-7 animals per timepoint. The sections were stained using the Histogene dye and roughly 500-1000 cells per gastric epithelial cell type were harvested by laser microdissection. RNA was prepared from these samples, subjected to two rounds of amplification, labelled and hybridized to 40.000 element murine cDNA arrays. The array names are composed as follows: "LCM" stands for laser capture microdissection; "mock" or "SS" stands for mock-infected or SS1-infected, respectively; "2,7,14 and 28" refers to the length of infection; "1-4" corresponds to the numbers given to every animal.The cellular origin of the RNA is represented by "mucus producing" or pit cell, "parietal" or acid-producing cell and "chief" or zymogenic cell. Between 5 and 7 samples were harvested per timepoint and cell type. Where the first labelling reaction and hybridization did not produce sufficiently good data (less than half of all spots made a regression correlation cutoff of >0.6), both were repeated (indicated by the addition of "relabeled" to the array name). The differential regulation of genes can be analyzed in both a time and cell type specific manner using publicly available software. Cell Type: "mucus producing" or pit cell (pit), acid-producing cell (parietal) and zymogenic cell (chief) time_series_design
2012-05-11 | E-GEOD-37938 | biostudies-arrayexpress
Project description:MIST1 is a bHLH transcription factor that is necessary for the maturation of gastric zymogenic cells as they differentiate from their precursor mucous neck cells. In this experiment, mucous neck cells and zymogenic cells of normal, adult C57BL/6 and MIST1 knockout mice were laser-capture microdissected in order to determine MIST1-dependent, zymogenic cell specific gene expression. Stomachs were excised immediately following sacrifice, quickly flushed with room-temperature PBS, inflated by duodenal injection of OCT compound, frozen in Cytocool II, and cut into serial 7-M-NM-<m-thick cryosections, which were mounted on Superfrost slides, fixed in 70% EtOH, rehydrated in nuclease-free water, and then incubated in Alexa Fluor 488-conjugated Griffonia simplicifolia GS-II (diluted 1:500 in nuclease-free water) for 15 min. Sections were washed in nuclease-free water and dehydrated in graded ethanol followed by xylene. ZCs were identified as corpus cells that were basal to GS-II labeling and which did not show the dark silhouettes and characteristic shape of parietal cells following xylene dehydration. Four wild-type mice and 5 Mist1M-bM-^HM-^R/M-bM-^HM-^R mice were used for dissection (PixCell II LCM apparatus [7.5-M-NM-<m spot diameter] and CapSure HS LCM caps) to generate two caps per mouse. RNA was purified by PicoPure kit, and RNA integrity was confirmed by an Agilent 2100 Bioanalyzer. All RNA from each cap was treated with DNase I and then reverse transcribed using the SuperScript III (Invitrogen) standard protocol (most cDNA syntheses started with 10 ng of total RNA). Biotinylated cRNA probes were hybridized to the GeneChips. Gene chip arrays used in these experiments were Affymetrix Mouse Gene 1.0ST arrays. Chip quality control and GeneChip to GeneChip comparisons to generate expression profiles were performed using dChip.
2013-01-12 | E-GEOD-43441 | biostudies-arrayexpress
Project description:MIST1 over-expression has not been analyzed in a cell type that does not express endogenous MIST1. Here, we force-express ectopic MIST1 in the parietal cells of mouse stomachs. We extracted whole corpus stomach mRNA and compared gene expression levels between MIST1-expressing mouse stomachs to controls.
2016-11-01 | GSE81549 | GEO
Project description:Mice were infected with Helicobacter pylori strain SS1 for 2, 7, 14 or 28 days or left uninfected. They were sacrificed at the appropriate times, the stomachs were removed and one half was plated for colony counts. The other half was embedded in OCT and cryosections were produced for 6-7 animals per timepoint. The sections were stained using the Histogene dye and roughly 500-1000 cells per gastric epithelial cell type were harvested by laser microdissection. RNA was prepared from these samples, subjected to two rounds of amplification, labelled and hybridized to 40.000 element murine cDNA arrays. The array names are composed as follows: "LCM" stands for laser capture microdissection; "mock" or "SS" stands for mock-infected or SS1-infected, respectively; "2,7,14 and 28" refers to the length of infection; "1-4" corresponds to the numbers given to every animal.The cellular origin of the RNA is represented by "mucus producing" or pit cell, "parietal" or acid-producing cell and "chief" or zymogenic cell. Between 5 and 7 samples were harvested per timepoint and cell type. Where the first labelling reaction and hybridization did not produce sufficiently good data (less than half of all spots made a regression correlation cutoff of >0.6), both were repeated (indicated by the addition of "relabeled" to the array name). The differential regulation of genes can be analyzed in both a time and cell type specific manner using publicly available software. Cell Type: "mucus producing" or pit cell (pit), acid-producing cell (parietal) and zymogenic cell (chief)
2012-05-12 | GSE37938 | GEO