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. 2011 Mar 16;43(5):237-54.
doi: 10.1152/physiolgenomics.00193.2010. Epub 2010 Dec 21.

Selective gene expression by rat gastric corpus epithelium

M Goebel  1 , A Stengel, N W G Lambrecht, G Sachs
Affiliations

Selective gene expression by rat gastric corpus epithelium

M Goebel et al. Physiol Genomics. .

Abstract

The gastrointestinal (GI) tract is divided into several segments that have distinct functional properties, largely absorptive. The gastric corpus is the only segment thought of as largely secretory. Microarray hybridization of the gastric corpus mucosal epithelial cells was used to compare gene expression with other segments of the columnar GI tract followed by statistical data subtraction to identify genes selectively expressed by the rat gastric corpus mucosa. This provides a means of identifying less obvious specific functions of the corpus in addition to its secretion-related genes. For example, important properties found by this GI tract comparative transcriptome reflect the energy demand of acid secretion, a role in lipid metabolism, the large variety of resident neuroendocrine cells, responses to damaging agents and transcription factors defining differentiation of its epithelium. In terms of overlap of gastric corpus genes with the rest of the GI tract, the distal small bowel appears to express many of the gastric corpus genes in contrast to proximal small and large bowel. This differential map of gene expression by the gastric corpus epithelium will allow a more detailed description of major properties of the gastric corpus and may lead to the discovery of gastric corpus cell differentiation genes and those mis-regulated in gastric carcinomas.

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Figures

Fig. 1.
Fig. 1.
Gene validation by RT-qPCR of selected genes. Based on microarray gene expression data 4 genes previously known to be selectively expressed in 1 segment (A) were chosen and mRNA expression assessed by RT-qPCR. Significant enrichment (log ratio >1) was found for colipase in gastric epithelial cells, intestinal alkaline phosphatase in proximal small bowel, matrilysin in distal small bowel, and mucosal pentraxin in colonic epithelial cells (B). C: integrity of RNA of gastric, proximal small bowel, distal small bowel, and colon epithelial cell isolations was determined by the electrophoretic trace of the RNA sample. The software algorithm allows for the classification of eukaryotic total RNA, based on a numbering system from 1 to 10, with 1 being the most degraded profile and 10 being the most intact. RNA integrity number (RIN) for the four samples was 9.9–10.0. D: 3 additional, novel genes were validated by RT-qPCR and significant mRNA expression of carbonic anhydrases (CAH) 9, CAH11, and the flippase Atp10d was found in the gastric corpus epithelium. Abbreviations: DSB, distal small bowel; PSB, proximal small bowel. ***P < 0.001 vs. all other groups, ##P < 0.01 vs. stomach and proximal small bowel.
Fig. 2.
Fig. 2.
Segment-specific expression of genes involved in acid secretion and pH homeostasis. This figure illustrates the relative distribution of genes encoding ATPases, bicarbonate transporters, and CAHs that are selectively expressed in epithelial cells of gastric corpus compared with PSB and DSB, or colon in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 3.
Fig. 3.
Protein expression of the flippase Atp10d in the gastric corpus mucosa and colocalization with gastric H,K-ATPase in parietal cells. Confocal microscopy of sections of rat gastric corpus doubly labeled with an antibody against Atp10d (red, A) and an antibody against the H,K-ATPase (green, B), showing that all Atp10d-immunoreactive cells in the gastric corpus colocalize with H,K-ATPase (C, D), a marker for parietal cells. The high-power confocal image (D) shows complete colocalization of this protein with the gastric H,K-ATPase within individual parietal cells (×100 objective, acquisition of fluorescent signals using multitract sequential scan).
Fig. 4.
Fig. 4.
Segment-specific expression of genes for ion and water channels. This figure illustrates the relative distribution of genes encoding potassium and chloride channels as well as aquaporins that are selectively expressed in epithelial cells of gastric corpus, compared with PSB, DSB, and colon in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 5.
Fig. 5.
Segment-specific expression of genes encoding endocrine mediators, growth factors, and receptors and binding proteins. This figure illustrates the relative distribution of genes encoding endocrine ligands, growth factors, and their receptors and binding proteins that are selectively expressed in epithelial cells of gastric corpus compared with the other GI segments in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 6.
Fig. 6.
Segment-specific expression of genes that code for proteins involved in epithelial protection, regeneration, and innate immune system. This figure illustrates the relative distribution of genes encoding proteins implicated in epithelial protection, regeneration, and innate immune system response that are selectively expressed in epithelial cells of the gastric corpus compared with PSB, DSB, and colon in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 7.
Fig. 7.
Segment-specific expression of genes coding for transcription factors. This figure illustrates the relative distribution of genes encoding transcription factors that are selectively expressed in epithelial cells of the gastric corpus in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 8.
Fig. 8.
Segment-specific expression of genes coding for various secreted digestive enzymes and involved in fatty acid, vitamin, and iron uptake. This figure illustrates the relative distribution of genes encoding digestive enzymes released into the lumen of different segments as well as proteins involved in uptake of fatty acids, vitamins, and iron in the GI tract that are selectively expressed in epithelial cells of gastric corpus compared with PSB and DSB, and colon in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 9.
Fig. 9.
Segment-specific expression of genes coding for various enzymes of glycolysis and glucose transporters. This figure illustrates the relative distribution of genes encoding various enzymes involved in glycolysis and glucose transport that are selectively expressed in epithelial cells of the gastric corpus in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
Fig. 10.
Fig. 10.
Segment-specific expression of genes coding for various enzymes involved in decarboxylation, alcohol dehydrogenation, and detoxification. This figure illustrates the relative distribution of genes encoding various enzymes involved in decarboxylation, alcohol dehydrogenation, and detoxification that are selectively expressed in epithelial cells of gastric corpus compared with PSB and DSB, or colon in 3 independent experiments (log of the mean of 3 ratios ± SD). Ratios are presented as heat maps showing gradients of highest (red) to lowest (green). The color scale at the bottom indicates the expression ratios (fold-change).
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