Author:

Keywords:

interstitial cells of cajal, dithp, prenylcysteine oxidase1, herne oxygenase2, phospholamban, troy, slow-transit constipation, actin-binding protein, heme oxygenase 2, w/w-v mice, interstitial-cells, gastrointestinal-tract, gastric fundus, c-kit, sarcoplasmic-reticulum, carbon-monoxide, Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Biophysics, interstitial cells of Cajal, Dithp, TROY, SLOW-TRANSIT CONSTIPATION, ACTIN-BINDING PROTEIN, HEME OXYGENASE 2, W/W-V MICE, INTERSTITIAL-CELLS, GASTROINTESTINAL-TRACT, GASTRIC FUNDUS, C-KIT, SARCOPLASMIC-RETICULUM, CARBON-MONOXIDE, Animals, COS Cells, Calcium-Binding Proteins, Carbon-Sulfur Lyases, Chlorocebus aethiops, Down-Regulation, Gene Expression Profiling, Heme Oxygenase (Decyclizing), Immunohistochemistry, Intestine, Small, Jejunum, Liver, Mice, Mice, Transgenic, Proto-Oncogene Proteins c-kit, Receptors, Tumor Necrosis Factor, Reverse Transcriptase Polymerase Chain Reaction, 0304 Medicinal and Biomolecular Chemistry, 0601 Biochemistry and Cell Biology, 1101 Medical Biochemistry and Metabolomics, 3101 Biochemistry and cell biology, 3404 Medicinal and biomolecular chemistry

Abstract:

Interstitial cells of Cajal (ICC) are the so-called pacemaker cells of the gut. W-LacZ/Wv and Sl/Sld mice lack ICC surrounding the myenteric plexus (MP) in the jejunum. We compared the gene expression profile of wild type (WT) and W-LacZ/Wv and Sl/Sld mice using suppression subtractive hybridization (SSH), generating a cDNA library of 1303 clones from which 48 unique sequences were differentially expressed with Southern blot. Among them, we identified heme oxygenase2, TROY, and phospholamban in ICC using immunohistochemistry. Using RT-qPCR, c-Kit and two new transcripts Dithp and prenylcysteine oxidase1 were significantly lower expressed in Sl/Sld and W-LacZ/Wv versus WT. Prenylcysteine oxidase1 appeared cytotoxic for COS-7 cells and was highly expressed in liver while Dithp was mainly expressed in small intestine. The combination of SSH, Southern blot, RT-qPCR, and immunohistochemistry turned out to be a useful approach to identify rarely expressed genes and genes with small differences in expression. (c) 2006 Elsevier Inc. All rights reserved.