Ultrastructural Detection of Neuronal Markers, Receptors, and Vesicular Transporters

Q2 Neuroscience Current Protocols in Neuroscience Pub Date : 2019-05-20 DOI:10.1002/cpns.70
Shiliang Zhang, Marisela Morales
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引用次数: 1

Abstract

At the ultrastructural level, axon terminals containing synaptic vesicles are clearly observed. These axon terminals (presynaptic component of a synapse) may be seen establishing contacts (synapses) with cell bodies, axons, or dendrites (postsynaptic component of a synapse). By a combination of ultrastructural analysis and immunodetection of molecules, it is possible to determine the subcellular distribution of specific cellular markers (i.e., enzymes), neurotransmitters (within synaptic vesicles), vesicular transporters (in association with vesicles), and receptors (within the presynaptic or postsynaptic component of a synapse). Here we will provide detailed protocols that facilitate the ultrastructural detection of cellular markers, receptors, and vesicular transporters. These protocols include brain ultrastructural immunodetection of one, two, or three different types of molecules prior to brain tissue processing for ultrastructural analysis (pre-embedding immunolabeling), brain molecular immunodetection after tissue processing for ultrastructural analysis (post-embedding immunolabeling), or molecular immunodetection in purified synaptic vesicles. Published 2019. This article is a US Government work and is in the public domain in the USA.

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神经元标记物、受体和囊泡转运体的超微结构检测
在超微结构水平上,可以清楚地观察到含有突触囊泡的轴突终末。这些轴突末端(突触的突触前成分)可以与细胞体、轴突或树突(突触的突触后成分)建立接触(突触)。通过结合超微结构分析和分子免疫检测,可以确定特定细胞标记物(即酶)、神经递质(在突触囊泡内)、囊泡转运蛋白(与囊泡相关)和受体(在突触的突触前或突触后成分内)的亚细胞分布。在这里,我们将提供详细的方案,以促进细胞标记物、受体和囊泡转运体的超微结构检测。这些方案包括在脑组织处理前进行一种、两种或三种不同类型分子的脑超微结构免疫检测(包埋前免疫标记),在组织处理后进行超微结构分析(包埋后免疫标记)的脑分子免疫检测,或在纯化的突触囊泡中进行分子免疫检测。2019年出版。这篇文章是美国政府的作品,在美国属于公有领域。
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Current Protocols in Neuroscience
Current Protocols in Neuroscience Neuroscience-Neuroscience (all)
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期刊介绍: Current Protocols in Neuroscience is a one-stop resource for finding and adapting the best models and methods for all types of neuroscience experiments. Updated every three months in all formats, CPNS is constantly evolving to keep pace with the very latest discoveries and developments. A year of these quarterly updates is included in the initial CPNS purchase price.
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