| Budget Amount *help |
¥7,000,000 (Direct Cost: ¥7,000,000)
Fiscal Year 1996: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥6,200,000 (Direct Cost: ¥6,200,000)
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| Research Abstract |
Glomerular mesangial cells (GMC) are thought to have a pivotal role in the development of glomerular diseases. The present study was undertaken to collect the gene expression profile of GMC,a database that describes which genes are expressed and to what extent. We used a 3'-directed regional cDNA library which was constructed from cultured human GMC by cleaving cDNA with Mbo Irestriction enzyme to avoid variable cloning efficiencies reflecting the size or base composition. We randomly selected 1197 clones from the library and sequenced them with an automatic DNA sequencer. The Sequence data were compared among the clones and also with the DNA databank (NCBI). We regarded two sequences as identical when they showed more than 90 percent homology. Among the 1197 sequenced clones, 450 clones appeared more than twice (representing 83 different species) and 747 clones appeared once or twice (representing 713 different species). Among 450 redundant clones, 344 clones corresponded to 53 genes,
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the functions of which are already known. When we classified these known genes according to their functions, a significant fraction of the genes expressed in human GMC are for secretable proteins (71 clones, 7 species) and for protein synthesis (74 clones, 11 species). The most abundant gene in human GMC is fibronectin (47 clones corresponding to 4.2 percent of total clones)/ A comparison of the expression profiles among different cells such as fibroblast, aortic endothelium, granulocyte, osteoblast, and HepG2, or tissues such as colon mucosa and lung, has allowed active genes to be classified as housekeepers or those with GMC cell-specific functions. We obtained 10 clones which were expressed especially in GMC,as compared to other cell types. By northern blot analysis using RNAs isolated from several human tissues, an intense signal for these clones were detected in kidney as well as cultured human GMC.One of 10 clones (GS9422) was shown to be a part of an unidentified gene with a homology to the serine protease inhibitor (serpin) family. In conclusion, the present study provide a database of genes expressed in human cultured GMC. Less
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