Serial microanalysis of renal transcriptomes

• DOI: 10.1073/pnas.96.26.15286
• Publication Date: 2002-07-26T14:39:15Z
• Authors: Bérangère Virlon; Lydie Cheval; Jean-Marie Buhler; Emmanuelle Billon; Alain Doucet; Jean-Marc Elalouf
• Keywords: Reference Genes

Large-scale gene expression studies can now be routinely performed on macroamounts of cells, but it is unclear to which extent current methods are valuable for analyzing complex tissues. In the present study, we used method serial analysis (SAGE) quantitative mRNA profiling in mouse kidney. We first SAGE at whole-kidney level by sequencing 12,000 tags. Most abundant tags corresponded transcripts widely distributed or enriched predominant kidney epithelial cells (proximal tubular cells), whereas specific minor cell types were barely evidenced. To better explore such set up a adaptation downsized extracts, enabling 1, 000-fold reduction amount starting material. The potential this approach was evaluated studying microdissected tubules (50,000 cells). Specific profiles obtained, and known markers (e.g., uromodulin thick ascending limb Henle's loop aquaporin-2 collecting duct) found appropriately enriched. addition, several had no databank match, suggesting that they correspond unknown poorly characterized with tissue distribution. It concluded extracts makes possible large-scale measurements small biological samples will help study function genes not evidenced other high-throughput methods.