In vivo imaging of axonal transport in murine motor and sensory neurons.
In vivo imaging of axonal transport in murine motor and sensory neurons.
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DOI：
10.1016/j.jneumeth.2015.09.018
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发表时间：
2016-01-15
影响因子：
3
通讯作者：
Schiavo G
中科院分区：
文献类型：
--
作者：
Gibbs KL;Kalmar B;Sleigh JN;Greensmith L;Schiavo G
活体麻醉小鼠坐骨神经轴突运输的活体成像。信号内体使用荧光探针和共聚焦显微镜进行监测。运动神经元和感觉神经元的轴突运输是可以区分的。这种方法可以很容易地适用于研究其他货物的轴突运输。这种体内成像方法在药物筛选中的潜在应用。轴突运输对神经元的功能和存活至关重要。轴突运输缺陷已被确认为几种神经系统疾病的早期病理特征。因此，神经疾病啮齿动物模型体内轴突运输的可视化和定量分析对于提高我们对疾病发病机制的理解和识别新的治疗方法是至关重要的。在这里，我们描述了一种活体成像的方法，用于在活的麻醉小鼠的坐骨神经中对信号内体的轴突运输进行成像。这种方法可以对成年小鼠在控制条件下和疾病进展过程中运动神经元和感觉神经元的体内轴突运输进行多参数、定量分析。以前对信号内体轴突运输的体内成像一直局限于神经植体准备或使用磁共振成像的非侵入性方法的研究；这些技术受到组织损伤和低时间和空间分辨率等主要缺陷的阻碍。这一新方法可以对坐骨神经中单个内小体的轴突运输进行现场成像，并比以前的方法更灵敏地分析轴突运输动力学。本文描述的方法可以深入分析活体运动神经元和感觉神经元的轴突运输特征。它能够详细研究神经疾病啮齿动物模型中轴突运输的变化，并可用于鉴定轴突运输的新的药理调节剂。
In vivo imaging of axonal transport in the sciatic nerve of live anesthetised mice. Signalling endosomes are monitored using fluorescent probes and confocal microscopy. Axonal transport in motor and sensory neurons can be differentiated. This method can be easily adapted to study the axonal transport of other cargoes. Potential use of this in vivo imaging approach in drug screening. Axonal transport is essential for neuronal function and survival. Defects in axonal transport have been identified as an early pathological feature in several disorders of the nervous system. The visualisation and quantitative analysis of axonal transport in vivo in rodent models of neurological disease is therefore crucial to improve our understanding of disease pathogenesis and for the identification of novel therapeutics. Here, we describe a method for the in vivo imaging of axonal transport of signalling endosomes in the sciatic nerve of live, anaesthetised mice. This method allows the multiparametric, quantitative analysis of in vivo axonal transport in motor and sensory neurons of adult mice in control conditions and during disease progression. Previous in vivo imaging of the axonal transport of signalling endosomes has been limited to studies in nerve explant preparations or non-invasive approaches using magnetic resonance imaging; techniques that are hampered by major drawbacks such as tissue damage and low temporal and spatial resolution. This new method allows live imaging of the axonal transport of single endosomes in the sciatic nerve in situ and a more sensitive analysis of axonal transport kinetics than previous approaches. The method described in this paper allows an in-depth analysis of the characteristics of axonal transport in both motor and sensory neurons in vivo. It enables the detailed study of alterations in axonal transport in rodent models of neurological diseases and can be used to identify novel pharmacological modifiers of axonal transport.