2000 Fiscal Year Final Research Report Summary
The analysis of genes related to the progressive kidney disease using gene expression progiles
| Project/Area Number |
11671035
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Kidney internal medicine
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| Research Institution | Osaka University |
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Principal Investigator |
TAKENAKA Masaru Osaka University Graduate School of Medicine, Assistant Professor, 医学系研究科, 助手 (20222101)
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| Co-Investigator(Kenkyū-buntansha) |
OOKUBO Kousaku Osaka University Institute for Molecular an Cellular Biology, Associate Professor, 細胞生体工学センター, 助教授 (40233069)
IMAI Enyu Osaka University Graduate School of Medicine, Lecture, 医学系研究科, 講師 (00223305)
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| Project Period (FY) |
1999 – 2000
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| Keywords | progressive renal disease / proteinuria / gene / database / laser microdissection / nephron |
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| Research Abstract |
The protein-overloaded proteinuira model has been reported to study renal damages caused by proteinuria, which we confirmed the same histological changes. We collected approximately 20cm mouse renal proximal tubules of disease model mice by micordissection methods under macroscopy, and extracted mRNA.The gene expression profiles were constructed by direct sequencing of 3' end of over 3000 cDNA clones that were randomly picked up from the transformants of un-amplified cDNA library. The profile contained the information about relative amount of mRNA expression. We compared those with gene expression profiles of normal tissues and cells. It revealed that the expression of genes related to proteolysis and inflammation were increased, whereas several abundant genes in normal proximal tubules were decreased, e.g. thymic shared antigen (TSA-1). Several genes were analyzed by Northern blot analysis, laser microdissection-real time PCR method, in situ hybridization and/or immunohistochemistry. It showed that mRNA expression of one of those genes, TSA-1, was increased in proximal tubular cells. Immunohistochemistry staining confirmed that its protein expression was a basolateral pattern and induced by proteinuria. The data of gene expression profile and microarray analysis revealed that osteopontin was induced by proteinuria in the kidney. When the informed consents were obtained, OPN mRNA expression in proximal tubules of renal biopsy specimen was quantified by the LMM method. The analyses revealed that OPN expression in proximal tubules was exponentially increased along with the amount of proteinuria.
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Research Products
(12 results)
