Mechanisms of regenerating spout formation and growth in the peripheral nerves as studied by confocal laser scanning and immunoelectron microscopy

通过共焦激光扫描和免疫电子显微镜研究周围神经再生喷口形成和生长的机制

• 批准号: 04454126
• 负责人: IDE Chizuka
• 金额: $ 3.84万
• 依托单位: Kobe University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04454126/
• 关键词: sprout; node of Ranvier; synaptophysin; Rab3A p25; protein kinase C; nerve regeneration; 再生軸索; 接着因子; N-カドヘリン; インテグリン; 小胞関連蛋白; 成長円錐; 軸索形質膜; プロテインキナーゼC; シナプシンI

1.The sprouting began as early as 3 hours after injury at the node of Ranvier. The nodal plasma membrane from which the electron dense undercoating had disappeared, protruded and further extended as regenerating axons through the space between the myelin sheath and basal lamina.2.The sprout formation as above occurred even when the node of Ranvier was disconnected from the cell body, indicating that the sprout formation is independent of effects from the cell body.3.The presence of synaptophysin was demonstrated in the sprouts at the node of Ranvier by immunoelectron microscopy, suggesting the possibility of exocytotic fusion of vesicles with the plasma membrane of the sprout.4.The localization of protein kinase C beta subtype was demonstrated in the axon terminal in the neuromuscular junction. Similarly, the subtypes (alpha, beta and gamma) were present in the sensory axon terminals of the muscle spindle.5.The localization of Rab3A p25 was demonstrated at the active zone of neuromuscular junction. In addition, the localization of Rabphilin-3A at the synaptic vesicles was demonstrated by immunogold method.6.The innervation processes of regenerating axons to the neuromuscular junction were demonstrated by PGP 9.5 immunohistochemistry.7.In addition to the above findings which were published in papers, the presence of synaptotagmin, synapsin I,N-cadherin and an integrin subtype were demonstrated in the growth cone of regenerating axons.

1.发芽早在Ranvier节点受伤后3小时开始。电子致密涂料的淋巴结质膜消失，突出并进一步延伸，作为通过髓鞘鞘和基底层之间的空间重新生成的轴突，即使在ranvier的节点与细胞的体中脱离了ranvier的节点，表明该细胞的形成是独立的。通过免疫电子显微镜在兰维尔节点的芽芽，这表明囊泡与发芽的质膜外胞囊融合的可能性。同样，在肌肉纺锤体的感觉轴突末端存在亚型（alpha，beta和伽马）。此外，通过免疫胶质方法证明了rabphilin-3a在突触囊泡处的定位。6。通过PGP 9.5免疫组织化学证明，再生轴突对神经肌肉连接的支配过程证明了上述发现的附加结果。在再生轴突的生长锥中证明了亚型。

项目成果

Okajima,S.,他: "Synaptophysin immunocytochemistry in the regenerating sprouts from the nodes of Ranvier in injured rat sciatic nerve." Brain Research. 631. 133-136 (1993)

Okajima, S., et al.：“受损大鼠坐骨神经 Ranvier 结节再生中的突触素免疫细胞化学。”631. 133-136 (1993)。

Masutani M.et al.: "Localization of protein kinase C α,β and γ subspecies in sensory axon terminals of the rat musclo spindle." Journal of Neurocytology. 23. 811-819 (1994)

Masutani M.et al.：“大鼠肌纺锤体感觉轴突末端的蛋白激酶 C α、β 和 γ 亚种的定位”，《神经细胞学杂志》23. 811-819 (1994)。

Okajima S.et al.: "Synaptophysin immunocytochemistry in the regenerating sprouts from the nodes of Ranvier in injured rat sciatic nerve." Brain Research. 631. 133-136 (1993)

Okajima S.et al.：“受损大鼠坐骨神经朗飞节再生芽中的突触素免疫细胞化学。”

Kato S.et al.: "Axonal Sprouting at the node of Ranvier of the peripheral nerve disconnected with the cell body" Restorative Neurology and Neuroscience. 6. 181-187 (1994)

Kato S.et al.：“与细胞体断开的周围神经朗飞节处的轴突发芽”恢复神经病学和神经科学。

Tomatsuri,M.,他: "Sprout formation at nodes of Ranvier of crush-injured peripheral nerves." Restorative Neurology and Neuroscience. 5. 275-282 (1993)

Tomatsuri, M., et al.：“挤压损伤的周围神经的 Ranvier 节点处的芽形成。”恢复神经学和神经科学 5. 275-282 (1993)