DBT-National Brain Research Centre;
J Vis Exp. 2021 Nov 20(177). doi: 10.3791/62744.
In neurons, microtubule orientation has been a key assessor to identify axons that have plus-end out microtubules and dendrites that generally have mixed orientation. Here we describe methods to label, image, and analyze the microtubule dynamics and growth during the development and regeneration of touch neurons in C. elegans. Using genetically encoded fluorescent reporters of microtubule tips, we imaged the axonal microtubules. The local changes in microtubule behavior that initiates axon regeneration following axotomy can be quantified using this protocol. This assay is adaptable to other neurons and genetic backgrounds to investigate the regulation of microtubule dynamics in various cellular processes.
在神经元中，微管取向一直是鉴定具有正极向外微管的轴突和通常具有混合取向的树突的关键评估器。在这里，我们描述了在秀丽隐杆线虫的触觉神经元发育和再生过程中标记、成像和分析微管动力学和生长的方法。我们使用微管尖端的遗传编码荧光报告子来成像轴突微管。使用该方案可以定量分析轴突切断后引发轴突再生的微管行为的局部变化。该测定法可适应于其他神经元和遗传背景，以研究各种细胞过程中微管动力学的调节。
