ELUCIDATION OF THE MAINTENANCE MECHANISM OF SYNAPTIC STRUCTURE AND FUNCTION BY THE PROTEIN QUALTY CONTROL SYSTEM AND ITS APPLICATION TO NEUROREGENERATION

蛋白质质量控​​制系统对突触结构和功能的维持机制的阐明及其在神经再生中的应用

• 批准号: 13480262
• 负责人: WADA Keiji
• 金额: $ 8.19万
• 依托单位: National Center of Neurology and Psychiatry (NCNP)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13480262/
• 关键词: ubiquitin; neurodegeneration; mouse; protein degradation; neuron; neurotransmission; neuroregeneration; synapse; 幹細胞; 神経軸索; パーキンソン病; 神経細胞死

We previously identified that ubiquitin C-terminal hydrolase L1 (UCH-L1) was not expressed in the gracile axonal dystrophy (gad) mouse. The gad mouse is pathologically characterized by dying-back type of axonal degeneration, and thus the mutant appears to be a suitable model for investigating the relationship between the ubiquitin system and synapes. In this study, we aimed to elucidate the physiological and pathophysiological roles of UCH-L1 in the maintenance of synaptic structure and function. We first identified that UCH-L1 unexpectedly bound to and stabilized monoubiquitin in neurons. In the stablization, hydrolase activity of UCH-L1 was not required. In the gad mouse, monoubiquitin level was decreased. These results suggest that UCH-L1 may function in multiple ways : as a hydrolase and as a ubiquitin-binding protein in vivo. Second, by proteaome analysis, we observed that four proteins were highly oxidized in the gad mouse. We also demonstrated that UCH-L1 itself was oxidized and decreased its hydrolase activity as the oxidation increased. Third, we observed that UCH-L1 played some role in synaptic plasticity. Since UCH-L1 is selectively expressed in neurons, our results suggest that UCH-L1 may play an essential role in synaptic transmission.

我们先前鉴定出泛素C末端水解酶L1（UCH-L1）未在尖轴轴突营养不良（GAD）小鼠中表达。 GAD小鼠在病理上以垂直的轴突变性类型进行病理学来表征，因此突变体似乎是研究泛素系统与突触之间关系的合适模型。在这项研究中，我们旨在阐明UCH-L1在维持突触结构和功能中的生理和病理生理作用。我们首先确定UCH-L1意外地与神经元中的单未木蛋白结合并稳定。在稳定化中，不需要UCH-L1的水解酶活性。在GAD小鼠中，单喹素水平降低。这些结果表明，UCH-L1可能以多种方式起作用：作为水解酶和体内的泛素结合蛋白。其次，通过蛋白质组分析，我们观察到在GAD小鼠中高度氧化了四种蛋白质。我们还证明，随着氧化的增加，UCH-L1本身被氧化并降低了其水解酶活性。第三，我们观察到UCH-L1在突触可塑性中发挥了一定作用。由于UCH-L1在神经元中有选择地表达，因此我们的结果表明UCH-L1可能在突触传播中起着至关重要的作用。

项目成果

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大阪 H.、王 Y.L.、高田 K.、泷泽 S.、节家 R.、李 H.、佐藤 Y.、西川 K.、孙 Y.J.、樱井 M.、原田，

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Castegna, A.、Thongboonkerd, V.、Klein, J.、Lynn, B.、Wang, Y.L.、Osaka, H.、Wada, K.、Butterfield, D.A.：“薄弱轴突营养不良症中脑蛋白的蛋白质组学分析（

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Nishikawa，K 等人：“帕金森病相关的人泛素羧基末端水解酶 L1 变体的结构和水解酶活性的改变”Biochem.Res.（出版中）。

Castegna, A.at al.: "Proteomic analysis of brain proteins in the gracile axonal dystrophy (gad) mouse, --"Neurochem.. 88・6. 1540-1546 (2004)

Castegna, A. 等：“薄弱轴突营养不良 (gad) 小鼠脑蛋白的蛋白质组学分析，--”Neurochem.. 88・6 (2004)。

Harada, T., Harada, C., Wang, Y.L., Osaka, H., Amanai, K., Tanaka, K., Takizawa, K., Setsuie, R., Sakurai, M., Sato, Y., Noda, M., Wada, K.: "Role of ubiquitin carboxy terminal hydrolase-LI in neural cell apoptosis induced by ischemic retinal injury in vi

原田 T.、原田 C.、王 Y.L.、大阪 H.、天内 K.、田中 K.、泷泽 K.、节家 R.、樱井 M.、佐藤 Y.、野田