中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2013, Vol. 8 ›› Issue (14): 1253-1261.doi: 10.3969/j.issn.1673-5374.2013.14.001

• 原著:神经损伤修复保护与再生 •    下一篇

不同示踪剂神经内注射对神经功能的影响

  

  • 收稿日期:2013-02-20 修回日期:2013-04-22 出版日期:2013-05-15 发布日期:2013-05-15

Neurological function following intra-neural injection of fluorescent neuronal tracers in rats

Wen Hu1, 2, Dan Liu2, 3, Yanping Zhang2, 3, Zhongyi Shen2, 4, Tianwen Gu2, 4, Xiaosong Gu1, 2, Jianhui Gu3   

  1. 1 School of Biology and Basic Medical Sciences, Soochow University, Suzhou 215123, Jiangsu Province, China
    2 Jiangsu Key Laboratory of Neuroregeneration, Nantong University, Nantong 226001, Jiangsu Province, China
    3 Department of Hand Surgery, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China
    4 School of Medicine, Nantong University, Nantong 226001, Jiangsu Province, China
  • Received:2013-02-20 Revised:2013-04-22 Online:2013-05-15 Published:2013-05-15
  • Contact: Jianhui Gu, Ph.D., Department of Hand Surgery, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China, 13809081785@163.com.
  • About author:Wen Hu☆, Studying for doctorate. Wen Hu and Dan Liu contributed equally to this work.

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摘要：

示踪剂在进行长时程示踪标记时必须满足低神经毒性的要求。以往虽然有一些神经示踪剂毒性的研究，但侧重于病理学角度，而应用示踪剂是否影响神经自身的功能尚不清楚。鉴于此，实验建立大鼠胫神经注射示踪剂模型，采用的神经示踪剂为5% Fluoro-Gold、4% True blue和10% Fluoro-Ruby，建模后采用足迹试验、痛觉评定、激光多普勒血流灌注成像、靶肌湿质量比检测及组织学染色等方法全面评价示踪后大鼠的运动、感觉和植物神经功能。结果显示，大鼠注射Fluoro-Gold和True blue可致后足皮肤血流灌注量显著增加，引起神经性血管扩张，而注射Fluoro-Ruby无影响。注射Fluoro-Gold大鼠后足底感觉功能减低，胫神经运动功能完全丧失，注射点远侧有髓神经纤维全部变性，神经纤维境界不清；注射True blue大鼠后足运动功能减退和有髓神经纤维变性程度较低。注射Fluoro-Ruby后，大鼠胫神经的运动和感觉功能无明显变化，有髓神经纤维未出现变性。结果证实，大鼠胫神经内注射Fluoro-Gold示踪剂可显著抑制运动、感觉和植物神经功能，注射True blue示踪剂次之，而注射Fluoro-Ruby示踪剂基本对神经功能无影响。

关键词: 神经再生, 周围神经损伤, 神经示踪, 毒性, 荧光剂, Fluoro-Gold, True Blue, Fluoro-Ruby, 胫神经, 感觉神经功能, 运动神经功能, 植物神经功能, 基金资助文章

Abstract:

Fluorescent neuronal tracers should not be toxic to the nervous system when used in long-term labeling. Previous studies have addressed tracer toxicity, but whether tracers injected into an intact nerve result in functional impairment remains to be elucidated. In the present study, we examined the functions of motor, sensory and autonomic nerves following the application of 5% Fluoro-Gold, 4% True Blue and 10% Fluoro-Ruby (5 μL) to rat tibial nerves via pressure injection. A set of evaluation methods including walking track analysis, plantar test and laser Doppler perfusion imaging was used to determine the action of the fluorescent neuronal tracers. Additionally, nerve pathology and ratio of muscle wet weight were also observed. Results showed that injection of Fluoro-Gold significantly resulted in loss of motor nerve function, lower plantar sensibility, increasing blood flow volume and higher neurogenic vasodilatation. Myelinated nerve fiber degeneration, unclear boundaries in nerve fibers and high retrograde labeling efficacy were observed in the Fluoro-Gold group. The True Blue group also showed obvious neurogenic vasodilatation, but less severe loss of motor function and degeneration, and fewer labeled motor neurons were found compared with the Fluoro-Gold group. No anomalies of motor and sensory nerve function and no myelinated nerve fiber degeneration were observed in the Fluoro-Ruby group. Experimental findings indicate that Fluoro-Gold tracing could lead to significant functional impairment of motor, sensory and autonomic nerves, while functional impairment was less severe following True Blue tracing. Fluoro-Ruby injection appears to have no effect on neurological function.

Key words: neural regeneration, peripheral nerve injury, neuronal tracing, tracer toxicity, neurological function, Fluoro-Gold, True Blue, Fluoro-Ruby, grants-supported paper, neuroregeneration