霍乱毒素B亚基与荧光金标记小鼠脊髓感觉神经和运动神经元的差异及量化分析
首发时间:2019-04-12
摘要:目的:比较霍乱毒素B亚基(cholera toxin B subunit, CTB)与荧光金(Fluoro-Gold, FG)用于标记小鼠脊髓损伤(Spinal Cord Injury, SCI)前后脊髓本体感觉神经和运动神经元的差异。方法:通过微量注射器将CTB-Alexa 488与FG分别注射到未损伤小鼠以及SCI小鼠的胫骨前肌(Tibialis Anterior, TA),通过免疫组织化学方法检测标记到的脊髓本体感觉神经和运动神经元,并用Imaris软件对标记到的脊髓本体感觉神经和运动神经元进行3D建模,定量分析其结构和体积。结果:在脊髓未损伤小鼠脊髓腰段,CTB-Alexa 488与FG均可以标记到运动神经元胞体,但CTB-Alexa 488可以标记到更广泛的运动神经元突起,且标记到的运动神经元胞体的体积更大。在SCI小鼠中,CTB-Alexa 488与FG标记的运动神经元突起的长度和数量没有显著差异,而CTB-Alexa 488标记的运动神经元胞体的体积显著大于FG标记的运动神经元胞体的体积。相比于未损伤小鼠,SCI小鼠中,通过CTB-Alexa 488标记可观察到运动神经元胞体的体积减小、突起数量减少和长度减短,以及本体感觉神经轴突的体积减小、轴突末梢的数量减少,而通过FG标记并不能区分这些变化。结论:相较于FG,CTB-Alexa 488标记能鉴别出小鼠脊髓损伤前后脊髓本体感觉神经和运动神经元重要的形态学改变,因此可以在小鼠脊髓损伤后更好的标记脊髓局部环路。
关键词: 神经生物学 霍乱毒素B亚基 荧光金 感觉神经和运动神经元 Imaris软件
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Difference and quantitative analysis of cholera toxin B subunit and Fluoro-Gold labeled spinal sensory nerve and motor neurons in mice
Abstract:Objective: To compare the differences between the cholera toxin B subunit (CTB) and Fluoro-Gold (FG) labeled spinal sensory nerve and motor neurons before and after spinal cord injury (SCI) in mice. Methods: Either CTB-Alexa 488 or FG was injected into tibialis anterior (TA) muscle of the un-injured mice and SCI mice by microinjector. The labeled spinal proprioceptive nerves and motor neurons were detected by immunohistochemistry, and then, 3D models were constructed by Imaris software for quantitatively analyzing the structure and volume of tracing spinal proprioceptive nerves and motor neurons. Results: The somas of the lumbar motor neurons could be labeled by both CTB-Alexa 488 and FG in the un-injured mice. Compared with FG tracing, more and longer neurites of motor neurons and larger volumes of motor neurons somas were labeled by CTB tracing. In SCI mice, there was no significant difference in the number and length of neurites of motor neurons by CTB and FG tracing, whereas the soma volumes of CTB labeled motor neurons were significantly larger than those of FG labeled motor neurons. As compared to the un-injured mice, using CTB tracing, we observed the decrease in volumes of motor neurons, decreased number and shortened length of neurites, decreased volumes of proprioceptive axons and axon terminals in SCI mice. However, these changes could not be observed by FG tracing. Conclusion: Compared with FG tracing, we can distinguish important changes of spinal proprioceptive nerves and motor neurons using CTB tracing before and after spinal cord injury in mice. Thus, CTB tracing of spinal proprioceptive nerves and motor neurons is a better choice to quantitatively analyze changes of spinal neural circuits after spinal cord injury in mice.
Keywords: Neurobiology Cholera toxin B subunit Fluoro-Gold sensory nerves and motor neurons Imaris software
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霍乱毒素B亚基与荧光金标记小鼠脊髓感觉神经和运动神经元的差异及量化分析
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