Transcription profiling of rat gastric ECL cells during 24 hrs fasting

ABSTRACT: Gastric ECL cells and parietal (PC) cells in normal conditions and gastric ECL cells after 24 hrs fasting were purified using a combination of counter-flow elutriation, nycodenz gradient and FACS based on acridine orange labeling of histamine containing vesicles (ECL cells) or autofluorescense (PC cells) to homogeneity. cRNA probes from this purified cell suspensions and from mixed gastric homogenates were hybridized to whole rat genome expression oligonucleotide microarrays. We imported each channel separately (normalized intensity of Cy3 labeled samples = gProcessedSignal and of Cy5 labeled samples = rProcessedSignal) from the original Agilent Feature Extraction data files (submitted as raw data) after slide scanning from all slides into a Genespring 7.3 microarray database warehouse. Genespring’s built in statistical module was used to compare all channels to each other. The normalization used was against specific samples (the 5 channels of Gastric mucosal scrapings). Genespring identifies genes differentially expressed in the ECL cell samples and the parietal samples and also calculates the significance of difference (p-value). Experiment Overall Design: ECL cells: free access to food and water (250 gm Sprague Dawley Rat) Experiment Overall Design: PC cells: free access to food and water (250 gm Sprague Dawley Rat) Experiment Overall Design: ECL cells: 24 hrs fasting, free access to water (250 gm Sprague Dawley Rat) Experiment Overall Design: Gastric mucosa scrapes: free access to food and water (250 gm Sprague Dawley Rat) Experiment Overall Design: Fasting gastric mucosa scrapes: 24 hrs fasting, free access to water (250 gm Sprague Dawley Rat) Experiment Overall Design: Three independent cell purifications, total RNA isolations and at least one dye swap per sample set (3).

ORGANISM(S): Rattus norvegicus

SUBMITTER: Nils Werner Georg Lambrecht 

PROVIDER: E-GEOD-6204 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress