中国神经再生研究(英文版) ›› 2022, Vol. 17 ›› Issue (5): 1034-1041.doi: 10.4103/1673-5374.324850

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

蜥蜴腰段横断脊髓中有限的轴突再生:生长相关蛋白43和神经丝免疫标记证据

  

  • 出版日期:2022-05-15 发布日期:2021-11-08

Growth associated protein 43 and neurofilament immunolabeling in the transected lumbar spinal cord of lizard indicates limited axonal regeneration

Lorenzo Alibardi*   

  1. Comparative Histolab Padova and Department of Biology, University of Bologna, Bologna, Italy
  • Online:2022-05-15 Published:2021-11-08
  • Contact: Lorenzo Alibardi, PhD, lorenzo. alibardi@unibo.it.

摘要:

以往对蜥蜴腰段横断脊髓的细胞学研究表明,脊髓近远残端之间桥区存在着分化的胶质细胞、少数神经元和轴突,有限数量的轴突(20-50个)可以穿过桥区,使脊髓近远残端小神经元重新建立连接,这种轴突再生似乎与最初瘫痪后后肢运动的恢复有关。为此,实验进一步扩展了以往的研究结果,发现蜥蜴腰段脊髓横断伤后11-34天形成了一个胶质连接组织桥,重新连接横断脊髓的近远侧残端。在最初瘫痪后,后肢运动功能的部分恢复发生在伤后1-3个月;48-50kDa生长相关蛋白43的免疫组化染色和超微结构分析表明,稀疏的生长相关蛋白43阳性轴突存在于脊髓近端残端,但其数量在伤后11-34天的组织桥中有所减少;伤后11-22天,少数200-220 kDa神经丝蛋白免疫染色阳性的轴突出现在组织桥中,且在伤后34天,进一步数量增加,至伤后3个月蜥蜴后肢运动功能已有所恢复,在脊髓的近远残端的许多神经元中生长相关蛋白43也出现阳性表达。这是一种转位到轴突生长锥的胞质蛋白,这表明生长相关蛋白43的合成与脊髓横断伤后神经元的轴突再生和萌发有关。有限的轴突重新连接在蜥蜴脊髓横断后1-3个月发生,桥内的少数再生轴突重新连接脊髓横断处近远残端,有助于后肢运动功能的恢复。

https://orcid.org/0000-0002-8247-2217 (Lorenzo Alibardi)

Abstract: Previous cytological studies on the transected lumbar spinal cord of lizards have shown the presence of differentiating glial cells, few neurons and axons in the bridge region between the proximal and distal stumps of the spinal cord in some cases. A limited number of axons (20–50) can cross the bridge and re-connect the caudal stump of the spinal cord with small neurons located in the rostral stump of the spinal cord. This axonal regeneration appears to be related to the recovery of hind-limb movements after initial paralysis. The present study extends previous studies and shows that after transection of the lumbar spinal cord in lizards, a glial-connective tissue bridge that reconnects the rostral and caudal stumps of the interrupted spinal cord is formed at 11–34 days post-injury. Following an initial paralysis some recovery of hindlimb movements occurs within 1–3 months post-injury. Immunohistochemical and ultrastructural analysis for a growth associated protein 43 (GAP-43) of 48–50 kDa shows that sparse GAP-43 positive axons are present in the proximal stump of the spinal cord but their number decreased in the bridge at 11–34 days post-transection. Few immunolabeled axons with a neurofilament protein of 200–220 kDa were seen in the bridge at 11–22 days post-transection but their number increased at 34 days and 3 months post-amputation in lizards that have recovered some hindlimb movements. Numerous neurons in the rostral and caudal stumps of the spinal cord were also labeled for GAP43, a cytoplasmic protein that is trans-located into their axonal growth cones. This indicates that GAP-43 biosynthesis is related to axonal regeneration and sprouting from neurons that were damaged by the transection. Taken together, previous studies that utilized tract-tracing technique to label the present observations confirm that a limited axonal re-connection of the transected spinal cord occurs 1–3 months post-injury in lizards. The few regenerating-sprouting axons within the bridge reconnect the caudal with the rostral stumps of the spinal cord, and likely contribute to activate the neural circuits that sustain the limited but important recovery of hind-limb movements after initial paralysis. The surgical procedures utilized in the study followed the regulations on animal care and experimental procedures under the Italian Guidelines (art. 5, DL 116/92).

Key words: GAP-43, immunocytochemistry, lizard, neurofilaments, regeneration, spinal cord

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