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.伤后3小时，郎维叶结开始发芽。电子致密底膜消失的节细胞质膜突出，并作为再生轴突穿过髓鞘和基底层之间的间隙进一步延伸。2.即使朗氏结与胞体分离，也能发生上述芽的形成。表明芽的形成不依赖于细胞体的作用。3.突触素的存在证明在芽的形成过程中，免疫电镜观察到囊泡与芽细胞质膜融合，提示囊泡与芽细胞质膜融合的可能性。4.蛋白激酶C β亚型定位于神经肌肉接头的轴突末端。Rab 3A p25在神经肌肉接头的活动区也有表达。此外，免疫胶体金法证实Rabphilin-3A在突触囊泡中的定位。6. PGP9.5荧光化学法证实再生轴突向神经肌肉接头的神经支配过程。7.除上述已发表的研究结果外，再生轴突生长锥中还发现了突触结合蛋白、突触蛋白I、N-钙粘蛋白和一种整合素亚型。

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)。

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。”

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)。

Tomatsuri S.et al.: "Sprouts formation at nodes of Ranvier of crush-injured peripheral nerve." Restorative Neurology and Neuroscience. 5. 275-282 (1993)

Tomatsuri S.et al.：“在挤压损伤的周围神经的 Ranvier 节点处形成芽。”

Arakawa M.et al.: "Ultrastructural localization of protein kinase C beta-subspecies in the axon terminal of rat neuromuscular junction." Neuroscience Research. 16. 125-130 (1993)

Arakawa M.et al.：“大鼠神经肌肉接头轴突末端蛋白激酶 C β 亚种的超微结构定位。”