Live imaging of microtubule dynamics at excitatory presynaptic boutons in primary hippocampal neurons and acute hippocampal slices.
Live imaging of microtubule dynamics at excitatory presynaptic boutons in primary hippocampal neurons and acute hippocampal slices.
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DOI：
10.1016/j.xpro.2021.100342
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发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Bartolini F
中科院分区：
文献类型：
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作者：
Qu X;Kumar A;Bartolini F
微管（MT）和谷氨酸能瞬时钮扣的末端动力学分析可以在小鼠或大鼠胚胎中分离的培养原代神经元中进行，也可以在体外电穿孔小鼠急性切片中进行。在这里，我们描述了一种方案，用于建立和分析表达MT +端结合蛋白EB3和突触前囊泡标记物vGlut1或VAMP2的标记版本的初级神经元和急性海马切片的实时图像记录。有关使用和执行本协议的完整信息，请参阅。培养神经元兴奋钮扣的MT +端动力学分析海马急性切片MT +端动力学分析培养神经元钮扣的MT +端动力学培养神经元钮扣的药理刺激培养神经元钮扣的MT +端动力学分析可以在小鼠或大鼠胚胎中分离的培养原代神经元中进行，也可以在小鼠子宫内电穿孔的急性切片中离体进行。在这里，我们描述了一种方案，用于建立和分析表达MT +端结合蛋白EB3和突触前囊泡标记物vGlut1或VAMP2的标记版本的初级神经元和急性海马切片的实时图像记录。
Analyses of microtubule (MT) plus end dynamics at glutamatergic en passant boutons can be carried out in cultured primary neurons isolated from mouse or rat embryos or ex vivo in acute slices isolated from mice that had been electroporated in utero. 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. Analysis of MT plus end dynamics at excitatory boutons in cultured neurons Analysis of MT plus end dynamics at excitatory boutons in acute hippocampal slices Pharmacological stimulation of MT nucleation at boutons in cultured neurons Analyses of microtubule (MT) plus end dynamics at glutamatergic en passant boutons can be carried out in cultured primary neurons isolated from mouse or rat embryos or ex vivo in acute slices isolated from mice that had been electroporated in utero. 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.