基础医学与临床 ›› 2018, Vol. 38 ›› Issue (7): 961-966.

• 研究论文 • 上一篇    下一篇

腺相关病毒与生物素葡聚糖胺顺行标记小鼠皮质脊髓束的比较

王坚,孙玉慧   

  1. 苏州大学
  • 收稿日期:2018-04-09 修回日期:2018-05-23 出版日期:2018-07-05 发布日期:2018-06-29
  • 通讯作者: 孙玉慧 E-mail:sunyuhui@suda.edu.cn
  • 基金资助:
    国家自然科学基金青年基金;江苏省自然科学基金青年基金

Comparison of anterograde tracing methods of mouse corticospinal tract with adeno associated virus and biotinylated dextran amine

  • Received:2018-04-09 Revised:2018-05-23 Online:2018-07-05 Published:2018-06-29

摘要: 目的 比较腺相关病毒(AAV)与生物素葡聚糖胺(BDA)用于顺行标记小鼠皮质脊髓束(CST)的差异,探索AAV病毒应用于小鼠皮质脊髓束的顺行神经示踪的优势。方法 通过立体定位注射将AAV8-CAG-GFP-2A病毒与BDA分别注射到正常小鼠以及脊髓损伤小鼠大脑体感运动皮层,通过免疫组织化学法检测AAV与BDA标记到的CST轴突,并对轴突束(bundle)及轴突分支(branch)的荧光强度进行定量分析。结果 在正常小鼠的脊髓颈段、胸段及脊髓损伤小鼠胸段,AAV与BDA均可标记到经典的CST轴突束,及投射到脊髓灰质中的轴突分支。相较于BDA,AAV在小鼠脊髓颈段和胸段均能标记到更广泛的CST轴突分支。结论 AAV对于CST轴突的标记效果优于传统的神经示踪剂BDA,可作为一种更好的神经示踪方法应用于小鼠皮质脊髓束及相关神经环路的形态学研究。

关键词: 腺相关病毒, 生物素葡聚糖胺, 皮质脊髓束

Abstract: Objective To compare the differences between Adeno Associated Virus (AAV) and Biotinylated Dextran Amine (BDA) in anterograde tracing of mouse corticospinal tract (CST), and to explore the advantages of the application of AAV in anterograde tracing of mouse CST axons as a neural tracer. Methods AAV8-CAG-GFP-2A and BDA were stereotactically injected into the mouse sensorimotor cortex of normal and spinal cord injury mice respectively, and immunohistochemistry was employed to observe the distribution of the labeled CST axons. The relative fluorescence of axon bundles and branches was analyzed. Results It was shown that both AAV and BDA could label the bundles of corticospinal tract and surrounding axon branches projecting into the gray matter of cervical and thoracic cords in both coronal and saggital sections of normal and spinal cord injury mice. Meanwhile, more axon branches were labeled in the AAV injected mice compared with those in the BDA injected ones. Conclusions AAV exhibits more superior performance in mouse CST axon tracing in comparison with BDA, and can be applied as a better neural tracer for the morphological study of mouse CST and related neural circuits.

Key words: Adeno Associated Virus, Biotinylated Dextran Amine, corticospinal tract

中图分类号: