Research on proteases related to demyelination expressed by oligodendrocytes

少突胶质细胞表达的脱髓鞘相关蛋白酶的研究

• 批准号: 13680830
• 负责人: YOSHIDA Shigetaka
• 金额: $ 1.6万
• 依托单位: Asahikawa Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680830/
• 关键词: myelin; spinal cord injury; proteases; mouse; neuropsin; EAE; oligodendrocyte; protease M; セリンプロテアーゼ; L1; 脱髄; 免疫組織化学

Spinal cord injury is medical and social problem because in most cases the recovery is poor. The limited recovery is mostly caused by scant potential of regeneration of the injured axons. Recent progress of research has shown that the extracellular environment, particularly myelin proteins, may be the primary factor for the inhibition of regeneration. We have shown that oligodendrocytes express extracellular proteases after injury to the central nervous system. In the current study we observed the change of expression of neuropsin and protease M after spinal cord injury and experimental allergic encephalitis (EAE), animal model of multiple sclerosis (MS). Furthermore, the change of possible substrates of neuropsin and nerve regeneration were also studied.All the experiments were approved by the Institute's Animal Ethical Committee.Untreated control mice showed the expression of neuropsin mRNA limited to motoneurons. Protease M mRNA was broadly expressed in white matter. After injury to … More the spinal cord, neuropsin mRNA was expressed by cells in the ventral and lateral funiculi. The upregulation was observed at 1-14 days after injury peaking at 4 day. Protease M protein was abundantly expressed in the lower CNS and the expression was colocalized with CNPase immunoreactivity, marker of mature oligodendrocytes. Electron microscopic analysis revealed that myelins were immunoreactive as well as oligodendrocyte cell bodies. After kainic acid injury, the immunoreactivity to protease M in myelin sheaths was stronger. To explore possible substrate for neuropsin, myelin protein was incubated with neuropsin and separated by electrophoresis and immunoblot was performed. The bands corresponding to myelin basic protein (MBP) became weakened after incubation with neuropsin and immunoblot analysis confirmed that these bands were MBP. EAE peaked 3-4 weeks after injection of MOG. With hematoxylin-eosin staining, infiltrating cells were observed subpial regions of the spinal cord. Neuropsin and protease M mRNA was expressed by the cell around the inflammatory regions.These results show the importance of extracellular proteases in physiological and pathological condition of the CNS. Less

脊髓损伤是一个医学和社会问题，因为在大多数情况下，恢复很差。有限的恢复主要是由于受损轴突的再生潜力不足。最近的研究表明，细胞外环境，特别是髓鞘蛋白，可能是抑制再生的主要因素。我们已经证明，少突胶质细胞表达细胞外蛋白酶损伤后，中枢神经系统。本研究观察了脊髓损伤、实验性变态反应性脑炎（EAE）、多发性硬化（MS）动物模型后神经鞘蛋白酶（neuropsin）和蛋白酶M（protease M）表达的变化。此外，我们还研究了neuropsin可能底物的变化和神经再生，所有实验都得到了该研究所动物伦理委员会的批准，未经处理的对照小鼠显示neuropsin mRNA的表达仅限于运动神经元。蛋白酶M mRNA在白色物质中广泛表达。受伤后， ...更多信息 在脊髓中，腹索和侧索中的细胞表达neuropsin mRNA。在损伤后1-14天观察到上调，在4天达到峰值。蛋白酶M蛋白在中枢神经系统下部大量表达，其表达与成熟少突胶质细胞的标志物-CN--免疫反应共定位。电子显微镜分析显示，髓鞘的免疫反应，以及少突胶质细胞体。海人酸损伤后，髓鞘中蛋白酶M的免疫反应增强。为了探索神经蛋白酶的可能底物，将髓鞘蛋白与神经蛋白酶孵育，并通过电泳和免疫印迹进行分离。髓鞘碱性蛋白（MBP）的条带在与neuropsin孵育后减弱，免疫印迹分析证实这些条带是MBP。EAE在注射MOG后3-4周达到高峰。苏木精-伊红染色可见脊髓软膜下有浸润细胞。结果表明，神经鞘蛋白酶和蛋白酶M mRNA在炎症区周围细胞表达，提示细胞外蛋白酶在中枢神经系统的生理和病理过程中起重要作用。少

项目成果

Kuwae, K.: "Epidermal expression of serine protease, neuropsin (KLK8) in normal and pathologic skin"Molecular Pathology. 55. 235-241 (2002)

Kuwae, K.：“正常和病理皮肤中丝氨酸蛋白酶、神经蛋白酶 (KLK8) 的表皮表达”分子病理学。

Shimizu-Okabe, C.: "L-isoform but not S-isoform of myelin associated glycoprotein promotes neurite outgrowth of mouse cerebellar neurons"Neurosci. Lett. 311. 203-205 (2001)

Shimizu-Okabe, C.：“髓磷脂相关糖蛋白的 L-异构体而非 S-异构体促进小鼠小脑神经元的神经突生长”Neurosci。

Yoshida, S.: "Extracellular environment and extracellular serine proteases in the central nervous system"Connective Tissue. (印刷中). (2003)

Yoshida, S.：“中枢神经系统中的细胞外环境和细胞外丝氨酸蛋白酶”结缔组织（出版中）。

Matsumoto-Miyai, K., Kitagawa, R., Ninomiya, A., Momota, Y., Yoshida, S., Matsumoto-Miyai, K., Kitagawa, R., Ninomiya, A., Momota, Y., Yosida, S., Shiosaka, S.: "Decidualization induces the expression and activation of an extracellular protease neuropsin

松本宫井，K.，北川，R.，二宫，A.，桃田，Y.，吉田，S.，松本宫井，K.，北川，R.，二宫，A.，桃田，Y.，吉田

Kuwae, K.: "Epidermal expression of serine protease, neuropsin (KLK8) in normal and pathologic skin"Molecular Pathology. (印刷中). (2002)

Kuwae, K.：“正常和病理皮肤中丝氨酸蛋白酶、神经蛋白酶 (KLK8) 的表皮表达”分子病理学（出版中）。