| Project/Area Number |
15590154
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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 |
General anatomy (including Histology/Embryology)
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| Research Institution | Gunma University |
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Principal Investigator |
AOKI Takeo Gunma University, Graduate School of Medicine, Assistant Professor, 大学院・医学系研究科, 講師 (70150919)
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| Co-Investigator(Kenkyū-buntansha) |
TAKATA Kuniaki Gunma University, Graduate School of Medicine, Professor, 大学院・医学系研究科, 教授 (20129290)
HAGIWARA Haruo Gunma University, Graduate School of Medicine, Associate Professor, 大学院・医学系研究科, 助教授 (80189464)
SUZUKI Takeshi Gunma University, Graduate School of Medicine, Assistant, 大学院・医学系研究科, 助手 (00261868)
MATSUZAKI Toshiyuki Gunma University, Graduate School of Medicine, Assistant, 大学院・医学系研究科, 助手 (30334113)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥2,400,000 (Direct Cost: ¥2,400,000)
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| Keywords | glucose transporter / translocation / rafts / caveolae endosome / endosome / clathrin / GTP binding proteins / GFP-fused protein expression / GFP融合蛋白 |
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| Research Abstract |
I made GFP fusion protein to have full length of wild type Caveolin-1(human) which was the protein which constituted caveolae toward and transfected into a 3T3-L1 cell and I got selected this by antibiotics stably expressed cells. I was able to get GFP fusion proteins stably expressed cells to have N terminal portion deleted type of this protein. In addition, N terminal deletion type of Caveolin-3 and Caveolin-1 DNA in two places of amino acids mutation of apart of scaffolding domain of Caveolin-1.
About a glucose transporter, I compared reaction for insulin stimulation after having guided fat cell differentiation with the 3T3-L1 cell which I transfected a GLUT4-GFP fusion type. As the factor which restrained the formation of caveolae, I transiently tranfected a N terminal portion deleted type (DGV) of Caveolin-3. As the factor which restrained the clathrin formation, I tranfected a C terminal part of AP180 DNA. I stimulated these two cells by insulin. As a result, a translocation of GLUT4 to a cell membrane was restrained by both protein expressions. This got a conclusion that both existence of clathrin coated vesicles and caveolae was restrained in a translocation to a cell membrane of a glucose transporter.
I examined a translocation and recycling of aquaporin-2 to compare it with a translocation of GLUT4. I understood that a translocation of aquaporin-2 was obstructed by functional restraint of raft caused by a defect of glycosphingolipid. Furthermore, after recycling, aquaporin retrieved endosomes with Caveolin-1. I understood what was obstructed return to this cell when I transfected caveolin-1 which had variation into two places of amino acids of a part of scaffolding domain which restrained the formation of caveolae.
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