中国神经再生研究(英文版) ›› 2015, Vol. 10 ›› Issue (10): 1700-1705.doi: 10.4103/1673-5374.167772

• 原著:脊髓损伤修复保护与再生 • 上一篇    下一篇

最有效标记大鼠脊髓运动神经元的逆行示踪剂

  

  • 收稿日期:2015-06-20 出版日期:2015-10-28 发布日期:2015-10-28
  • 基金资助:

    国家重点基础研究发展计划(973计划)(2014CB542200);教育部创新团队发展计划(IRT1201);国家自然科学基金项目(31271284,31171150,81171146,30971526,31040043,31371210,81372044,31471144),北京市科技基金(7142164)

Comparison of commonly used retrograde tracers in rat spinal motor neurons

You-lai Yu1, Hai-yan Li2, Pei-xun Zhang1, *, #, Xiao-feng Yin1, Na Han1, Yu-hui Kou1, Bao-guo Jiang1, *, #   

  1. 1 Department of Trauma and Orthopedics, Peking University People’s Hospital, Beijing, China
    2 Department of Gastroenterology, Weifang People’s Hospital, Weifang, Shandong Province, China
  • Received:2015-06-20 Online:2015-10-28 Published:2015-10-28
  • Contact: Bao-guo Jiang or Pei-xun Zhang,jiangbaoguo@vip.sina.com or zhangpeixun@126.com.
  • Supported by:

    This study was supported by a grants from the National Program on Key Basic Research Project (973 Program), No. 2014CB542200; the Innovative Research Team by the Ministry of Education, No. IRT1201; the National Natural Science Foundation of China, No. 31271284, 31171150, 81171146, 30971526, 31040043, 31371210, 81372044, 31471144; and the Beijing Municipal Natural Science Foundation of China, No. 7142164.

摘要:

为探索神经示踪剂真蓝(TB)、荧光金(FG)、羰化青(DiI)和红色荧光金(FR)对大鼠脊髓运动神经元标记效率的差异,实验以大鼠股神经肌支为示踪模型,以真蓝、荧光金、羰化青和红色荧光金中的一或两种浸泡股神经横断后近侧断端。单标实验中将股神经横断后近侧断端分别用真蓝、荧光金、羰化青和红色荧光金中的一种进行示踪,荧光显微镜观察到示踪术后3d,红色荧光金示踪的大鼠脊髓腰膨大段神经元的数量最少,但1周时4种示踪剂标记的神经元数量接近。双标实验中将荧光金联合羰化青或真蓝联合羰化青组合后示踪1周时,2种示踪剂组合标记的神经元数量接近,说明示踪剂真蓝和羰化青/荧光金能有效标记相同的神经元,作者认为在逆行示踪大鼠脊髓运动神经元的研究中,使用荧光金和羰化青组合或真蓝和羰化青组合是比较合适的选择。

关键词: 神经再生, 示踪效率, 荧光示踪剂, 逆行示踪, 运动神经元, 大鼠股神经

Abstract:

The purpose of this study was to investigate the effect of four fluorescent dyes, True Blue (TB), Fluoro-Gold (FG), Fluoro-Ruby (FR), and 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI), in retrograde tracing of rat spinal motor neurons. We transected the muscle branch of the rat femoral nerve and applied each tracer to the proximal stump in single labeling experiments, or combinations of tracers (FG-DiI and TB-DiI) in double labeling experiments.
In the single labeling experiments, significantly fewer labeled motor neurons were observed after FR labeling than after TB, FG, or DiI, 3 days after tracer application. By 1 week, there were no significant differences in the number of labeled neurons between the four groups. In the double-labeling experiment, the number of double-labeled neurons in the FG-DiI group was not significantly different from that in the TB-DiI group 1 week after tracer application. Our findings indicate that TB, FG, and DiI have similar labeling efficacies in the retrograde labeling of spinal motor neurons in the rat femoral nerve when used alone. Furthermore, combinations of DiI and TB or FG are similarly effective. Therefore, of the dyes studied, TB, FG and DiI, and combinations of DiI with TB or FG, are the most suitable for retrograde labeling studies of motor neurons in the rat femoral nerve.

Key words: nerve regeneration, tracing efficacy, fluorescent tracers, retrograde tracing, femoral nerve, motor neurons