Department of Pathology & Cell Biology, Columbia University Medical Center, New York, NY 10032, USA.
STAR Protoc. 2021 Feb 21;2(1):100342. doi: 10.1016/j.xpro.2021.100342. eCollection 2021 Mar 19.
Analyses of microtubule (MT) plus end dynamics at glutamatergic boutons can be carried out in cultured primary neurons isolated from mouse or rat embryos or in acute slices isolated from mice that had been electroporated . Here, we describe a protocol for setting up and analyzing live image recordings of primary neurons and acute hippocampal slices expressing tagged versions of the MT plus end binding protein EB3 and the presynaptic vesicle markers vGlut1 or VAMP2. For complete information on the use and execution of this protocol, please refer to Qu et al. (2019).
在培养的原代神经元或从已电穿孔的小鼠中分离的急性切片中,可以分析谷氨酸能末梢的微管(MT)末端动力学。 在这里,我们描述了一个方案,用于建立和分析表达标记的 MT 末端结合蛋白 EB3 和突触前囊泡标记物 vGlut1 或 VAMP2 的原代神经元和急性海马切片的活细胞图像记录。 有关此方案使用和执行的完整信息,请参见 Qu 等人(2019 年)。