| Project/Area Number |
15591216
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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 |
Psychiatric science
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| Research Institution | Hamamatsu University School of Medicine |
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Principal Investigator |
OHNO Koji Hamamatsu University School of Medicine, Department of Anatomy, Associate professor, 医学部, 助教授 (90263277)
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| Co-Investigator(Kenkyū-buntansha) |
MIKAWA Sumiko Hamamatsu University School of Medicine, Department of Anatomy, Assistant professor, 医学部, 助手 (70359743)
渡部 和男 浜松医科大学, 医学部, 助手 (60107828)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | monoamine / depression / transporter / screening / VMAT / 結合蛋白 / VMAT-2 / モノアミントランスポーター / yeast two-hybrid |
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| Research Abstract |
The onset of mental diseases such as depression is closely related with the disorder of monoaminergic nervous system. In the CNS, the vesicular monoamine transporter, VMAT-2, is localized at synaptic vesicles and plays a vital role in the storage of monoamine in synaptic vesicles. Therefore, VMAT-2 is essential to maintain normal monoaminergic transmission and it is conceivable that VMAT-2 is one of the therapeutic targets of mental diseases. In this respect, we wanted to know how VMAT-2 function is regulated in vivo. Recent studies have shown that VMAT-2 C-terminal involves a signal for determining the subcellular localization of VMAT-2, suggesting that protein interaction through C-terminal might regulate the function of VMAT-2. To test this hypothesis, we planned to search for proteins that interact with VMAT-2 C-terminal. To this end, we first performed yeast two-hybrid (Y2H) screening on rat brain cDNA library and the results of sequence analyses showed that that most of obtained colonies by Y2H included transcription-related gene cDNAs, which appeared false positive. To explore in vivo binding partners more efficiently, we next generated a cDNA library from the rat striatum, where VMAT-2 is highly expressed, and repeated Y2H screening. Finally we obtained 68 colonies, but unfortunately we could not read out interesting cDNAs from these colonies. Now we have changes the strategy and are trying to analyze VMAT-2 protein complexes directly by the combination of blue-native PAGE and MASS spectrometer. Although we could not finish this project according to our planning, we have still much interest in this subject and are planning to continue our investigation.
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