中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (5): 751-756.doi: 10.4103/1673-5374.206644

• 原著:神经损伤修复保护与再生 • 上一篇    下一篇

低能双光子纳米外科手术对活体中枢神经系统神经元结构破坏的动态变化

  

  • 收稿日期:2017-05-06 出版日期:2017-05-15 发布日期:2017-05-15
  • 基金资助:

     

    973项目；中国香港健康与医学研究基金

Time-lapse changes of in vivo injured neuronal substructures in the central nervous system after low energy two-photon nanosurgery

Zhikai Zhao1, Shuangxi Chen1, Yunhao Luo2, Jing Li1, Smaranda Badea2, Chaoran Ren1, Wutian Wu1, 2, 3, 4   

  1. 1 Guangdong-Hong Kong-Macau Institute of CNS Regeneration, Jinan University, Guangzhou, Guangdong Province, China; 2 School of Biomedical Sciences, Division of Anatomy, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China; 3 Guangdong Engineering Research Center of Stem Cell Storage and Clinical Application, Saliai Stem Cell Science and Technology, Guangzhou, Guangdong Province, China; 4 State Key Laboratory of Brain and Cognitive Sciences, The University of Hong Kong, Pokfulam, Hong Kong Special Administrative Region, China
  • Received:2017-05-06 Online:2017-05-15 Published:2017-05-15
  • Contact: Wutian Wu, M.D., Ph.D.,wtwu@hku.hk.
  • Supported by:

    This work was supported by National Basic Research Program of China (973 Program), No. 20114002002, and 2014CB542205, and by Hong Kong Health and Medical Research Fund, No. 02132826.

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摘要：

关于中枢神经系统损伤后神经元亚细胞结构退行性改变的动态研究目前仍十分少见。此次实验应用一种新型方法低能小于30 mW双光子纳米外科手术成功对活体小鼠大脑和脊髓神经元的单个树突、轴突和树突棘进行选择性破坏，且体内成像精确监测了这些损伤的神经元结构的时间推移变化。实验用的双光子能量很低，因而不能引起神经元亚结构，特别是树突棘的额外损伤；同时，低能双光子纳米外科手术进行过程中还能对神经元结构的损伤程度进行实时动态监测，因而这项神经显微外科技术有助于研究活体中枢神经系统损伤后神经元亚细胞结构的变化。

ORCID：0000-0002-0959-4664(Wutian Wu)

关键词: 神经再生, 树突, 轴突, 树突棘, 脑, 脊髓, 双光子纳米外科, 活体

Abstract:

There is currently very little research regarding the dynamics of the subcellular degenerative events that occur in the central nervous system in response to injury. To date, multi-photon excitation has been primarily used for imaging applications; however, it has been recently used to selectively disrupt neural structures in living animals. However, understanding the complicated processes and the essential underlying molecular pathways involved in these dynamic events is necessary for studying the underlying process that promotes neuronal regeneration. In this study, we introduced a novel method allowing in vivo use of low energy (less than 30 mW) two-photon nanosurgery to selectively disrupt individual dendrites, axons, and dendritic spines in the murine brain and spinal cord to accurately monitor the time-lapse changes in the injured neuronal structures. Individual axons, dendrites, and dendritic spines in the brain and spinal cord were successfully ablated and in vivo imaging revealed the time-lapse alterations in these structures in response to the two-photon nanosurgery induced lesion. The energy (less than 30 mW) used in this study was very low and caused no observable additional damage in the neuronal sub-structures that occur frequently, especially in dendritic spines, with current commonly used methods using high energy levels. In addition, our approach includes the option of monitoring the time-varying dynamics to control the degree of lesion. The method presented here may be used to provide new insight into the growth of axons and dendrites in response to acute injury.

Key words: nerve regeneration, dendrite, dendritic spine, axon, spinal cord, two-photon nanosurgery, single-synapse resolution, neural regeneration