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.

我们之前发现泛素c端水解酶L1 （UCH-L1）在细轴索营养不良（gad）小鼠中不表达。gad小鼠的病理特征是轴突变性的死退型，因此该突变体似乎是研究泛素系统与突触之间关系的合适模型。在本研究中，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。我们首先发现UCH-L1意外地结合并稳定了神经元中的单泛素。在稳定过程中，不需要UCH-L1的水解酶活性。在小鼠中，单泛素水平降低。这些结果表明UCH-L1可能以多种方式发挥作用：在体内作为水解酶和泛素结合蛋白。其次，通过蛋白质组分析，我们观察到四种蛋白质在小鼠体内被高度氧化。我们还证明UCH-L1本身被氧化，并随着氧化的增加而降低其水解酶活性。第三，我们观察到UCH-L1在突触可塑性中起一定作用。由于UCH-L1在神经元中选择性表达，我们的研究结果表明UCH-L1可能在突触传递中发挥重要作用。

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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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)。

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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）。

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