| Project/Area Number |
17590156
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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 | Ibaraki Prefectural University of Health Science (2006-2007)
University of Yamanashi (2005) |
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Principal Investigator |
BABA Takeshi Ibaraki Prefectural University of Health Science, Department of Health Sciences, Professor (90208710)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,800,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥300,000)
Fiscal Year 2007: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥1,500,000 (Direct Cost: ¥1,500,000)
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| Keywords | lipid raft / cholera toxin / lysenin / immunological synapse / ultrastructure / immunocvtochemistry / 免疫学的シナプ / コレステロール / 電子顕微鏡 / 免疫細胞化学 / 凍結レプリカ |
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| Research Abstract |
Recent studies have revealed that lipid composition of cell membranes are not uniform but heterogeneous mixture of various membrane microdomains such as lipid rafts. However, ultrastructure of cytoplasmic counterparts of those membrane microdomains on the cell surface and corresponding signal transducing molecules are not well understood.
In the present research period, we studied the ultrastructural distribution of various lipid raft markers and obtained the following results.
We successfully induced immunological synapse formation by incubating Jurkat cells with coverslips coated with anti-D3 antibody. Next, we studied the distribution of lipid raft markers during immunological synapse formation. The cells were incubated with 5m-colloidal gold-labeled lysenin and 10nm-colloidal gold-labeled cholera toxin at 4℃. Labeled cells were incubated with anti-D3 antibody-coated coverslips at 4℃, and warmed up to 37℃ to induce immunological synapse formation. Fluorescence microscopic observations using HV161-lysenin and Alexa594-labeled cholera toxin revealed that randomly distributed lipid raft markers at 4℃ were accumulated surrounding attached area after incubation at 37℃. Ultrastructural study have revealed that both marker were accumulated around the immunological synapse, but lysenin was also distributed on the attached area. These results suggested that redistribution of lipid rafts during immunological synapse formation differs according to lipid type.
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