| Project/Area Number |
09671165
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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 |
MORIYAMA Toshio Osaka University, School of Health and Sport Sciences, Assistant Professor, 健康体育部, 講師 (30283815)
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| Co-Investigator(Kenkyū-buntansha) |
MIWA Takeshi Osaka University, Genoma Information Research Center, Associate Professor, 遺伝実験情報施設, 助教授 (20174229)
IMAI Enyu Osaka University Medical School, Research Assistant, 医学部, 助手 (00223305)
ANDO Akio Osaka University, School of Health and Sport Sciences, Professor, 健康体育部, 教授 (00028656)
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| Project Period (FY) |
1997 – 1998
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| Project Status |
Completed (Fiscal Year 1998)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | in vivo promoter analysis / myofibroblast / habu-venom / glomerulonephritis / phenotypic modulation / GArG element / smooth muscle alpha actin / transgenic mouse |
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| Research Abstract |
Activation of glomerular mesangial cells is one of the early, important features of progressive glomerular disease.Smooth muscle alpha-actin (SMalphaA) is an excellent marker of activated mesangial cells.We examined in vivo promoter analysis of the SMalphaA gene utilizing transgenic mice harboring different promoter regions of the SMalphaA gene fused to chloramphenicol acetyl transferase (CAT).CAT activities were tested in primary cultured mesangial cells, and in glomerular legions of habu-venom glomerulonephritis.DNA sequence -891 to +3828, which contains exon 1, intron 1, and the first 14 bp of exon 2 in addition to the 5'-flanking sequence of the SMalphaA gene, induced high levels of transcription in activated mesangial cells in in vivo habu-venom glomerulonephritis and in cultured mesangial cells derived from transgenic mice.DNA region -891 to -124 was a positive element in mesangial cells derived from transgenic mice.Deletions (3316 bp or 137 bp) in intron 1 reduced transcription to undetectable levels.The 137 bp sequence is highly conserved among several species, containing one CArG box element, which is one of the key motifs for transcriptional activation of contractile-related proteins.In vitro transfection analysis failed to demonstrate these positive effects of intron 1 and region -891 to -124.Further analysis of the intron 1 region (+1088 to +1224), including the CArG box element, will be of great value in understanding the molecular mechanisms of mesangial activation.
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