| Project/Area Number |
24650186
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Nerve anatomy/Neuropathology
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| Research Institution | Kyushu University |
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Principal Investigator |
IWAKI Toru 九州大学, 医学(系)研究科(研究院), 教授 (40221098)
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| Research Collaborator |
KIDOAKI Satoru 九州大学, 先導物質化学研究所, 教授 (10336018)
HONDA Hiroyuki 九州大学, 大学院医学研究院, 助教 (90624057)
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| Project Period (FY) |
2012-04-01 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥3,770,000 (Direct Cost: ¥2,900,000、Indirect Cost: ¥870,000)
Fiscal Year 2014: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
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| Keywords | 脳神経疾患 / プリオン病 / 病理学 / 蛋白重合度 / 検出技術 / 全反射蛍光顕微鏡 / 痴呆 / 蛋白オリゴマー |
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| Outline of Final Research Achievements |
The degree of polymerization of PrP has a close relationship with the pathological mechanisms of prion diseases. We examined the polymerization state of PrP in lysates of prion-infected cells using total internal reflection fluorescence microscopy (TIRFM). The crude lysates were fractionated by gelfiltration. Both the oligomer-rich and the monomer-rich fractions were probed with fluorescein-labeled anti-PrP antibodies (mAb SAF70 and mAb 8G8). Fluorescent spots of varying intensity were detected, with the ratio of intense fluorescent spots being greater in the oligomer fraction samples with mAb SAF70 than those with 8G8, the specific epitope of which is thought to be buried in abnormal PrP molecules. The results indicated that PrP oligomers could be specifically detected and conformational changes of abnormal PrP molecules observed. Imaging by TIRFM may aid in determining the polymerization state and properties of PrP oligomers in pathological processes.
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