Nanoscale Molecular Imaging of Presynaptic Active Zone Proteins in Cultured Hippocampal Neurons
Nanoscale Molecular Imaging of Presynaptic Active Zone Proteins in Cultured Hippocampal Neurons
复制标题
培养海马神经元突触前活性区蛋白的纳米分子成像
DOI：
10.1007/978-1-0716-0532-5_12
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Hirose Kenzo
中科院分区：
文献类型：
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作者：
Sakamoto Hirokazu;Namiki Shigeyuki;Hirose Kenzo
神经递质的释放发生在神经末梢的一个特殊区域，称为突触前活动区，其中存在大量的突触蛋白。活性区蛋白的组成和纳米级空间组织是突触前功能的关键决定因素，并且组成和组织的多样性可能解释了中央突触的广泛多样性。在这里，我们描述了随机光学重建显微镜（STORM）成像，基于单分子定位的超分辨率成像，用于分析培养的海马神经元中活性区蛋白的纳米级分布。STORM能够以显著的空间分辨率进行三维和三维分析，从而解析由活性区蛋白形成的详细纳米结构，例如对应于量子突触囊泡释放位点的Munc 13 -1超分子纳米组装体。在本章中，我们将为读者提供有关如何对突触前活动区蛋白进行染色、准备适合STORM成像的标本以及以定量方式进行STORM成像的指南。
Neurotransmitter release occurs at a specialized region of the nerve terminal known as the presynaptic active zone, where a large number of synaptic proteins reside. The composition and nanoscale spatial organization of the active zone proteins are crucial determinants of presynaptic functions, and variety in the composition and organization may account for wide diversity of central synapses. Here, we describe stochastic optical reconstruction microscopy (STORM) imaging, a single-molecule localization-based super-resolution imaging, for analyzing nanoscale distributions of active zone proteins in cultured hippocampal neurons. STORM enables multicolor and three-dimensional analysis with remarkable spatial resolution, and thus resolves detailed nanostructures formed by the active zone proteins, such as Munc13-1 supramolecular nanoassemblies which correspond to the quantal synaptic vesicle release sites. In this chapter, we provide the reader with guidelines on how to stain presynaptic active zone proteins, to prepare specimens suitable for STORM imaging, and to perform STORM imaging in a quantitative manner.
影响因子：
16.6
作者：
Ehmann, Nadine;van de Linde, Sebastian;Alon, Amit;Ljaschenko, Dmitrij;Keung, Xi Zhen;Holm, Thorge;Rings, Annika;DiAntonio, Aaron;Hallermann, Stefan;Ashery, Uri;Heckmann, Manfred;Sauer, Markus;Kittel, Robert J.
通讯作者：
Kittel, Robert J.
影响因子：
64.8
作者：
通讯作者：
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