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

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。

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.