Downregulation of Two Novel Genes in Sl/Sld and W(Lacz)/Wv Mouse Jejunum.
From: Department of Internal Medicine, Johnson and Johnson, Pharmaceutical Research and Development, A Subdivision of Janssen Pharmaceutics, Beerse, Belgium.
Biochemical and biophysical research communications
- Publish Date: Jul 2006
- ISSN: 0006-291X
- Volume: 346
- Issue: 2
- Pages: 491-500
- Medium: Print
- Language: English
- Citation (JAMA): Wouters Mira M, Neefs Jean-Marc, Kerchove d'Exaerde Alban de, et al. Downregulation of Two Novel Genes in Sl/Sld and W(Lacz)/Wv Mouse Jejunum.. Biochem. Biophys. Res. Commun. Jul 2006;346:491-500
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.
Mesh Headings (Keywords): Animals, COS Cells, Calcium-Binding Proteins, Carbon-Sulfur Lyases, Cercopithecus 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
Check for Full Text / PubMed Unique Identifier (PMID): 16765319
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