Assessing Neuron-Astrocyte Spatial Interactions Using the Neuron-Astrocyte Proximity Assay.
Assessing Neuron-Astrocyte Spatial Interactions Using the Neuron-Astrocyte Proximity Assay.
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DOI：
10.1002/cpns.91
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发表时间：
2020-03-01
影响因子：
--
通讯作者：
Khakh, Baljit S
中科院分区：
文献类型：
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作者：
Badia-Soteras, Aina;Octeau, J Christopher;Khakh, Baljit S
星形胶质细胞是一种形态复杂的细胞，在其胞体、分支和小枝水平上与神经元有许多密切的联系。最小的星形胶质细胞过程与突触在标准光学显微镜无法观察到的尺度上进行离散接触。在这样的接触点，星形胶质细胞被认为是执行自我平衡和神经调节的作用，提出了由其紧密的空间并置决定的功能。为了研究这种空间相互作用，我们以前开发了一种基于福斯特共振能量转移（FRET）的方法，该方法可以观察和跟踪星形胶质细胞与突触过程的静态和动态接近。该方法与标准成像技术，如共聚焦显微镜兼容，并允许评估活组织中星形胶质细胞和神经元之间的最接近的接触。在这篇协议文章中，我们描述的方法来分析纹状体星形胶质细胞的过程和皮质纹状体神经元投射终端到中型棘神经元之间的联系。我们详细报告所需的协议，包括腺相关病毒显微注射，急性脑切片制备，成像和事后FRET定量。文章提供了一个详细的描述，可用于表征和研究星形胶质细胞的过程接近活组织中的突触。© 2020年由约翰威利父子，公司。基本方案1：培养细胞中的Forster共振能量转移成像基本方案2：急性脑切片中神经元-星形胶质细胞邻近测定的Forster共振能量转移成像。
Astrocytes are morphologically complex cells with numerous close contacts with neurons at the level of their somata, branches, and branchlets. The smallest astrocyte processes make discrete contacts with synapses at scales that cannot be observed by standard light microscopy. At such contact points, astrocytes are thought to perform both homeostatic and neuromodulatory roles-functions that are proposed to be determined by their close spatial apposition. To study such spatial interactions, we previously developed a Forster resonance energy transfer (FRET)-based approach, which enables observation and tracking of the static and dynamic proximity of astrocyte processes with synapses. The approach is compatible with standard imaging techniques such as confocal microscopy and permits assessment of the most proximate contacts between astrocytes and neurons in live tissues. In this protocol article we describe the approach to analyze the contacts between striatal astrocyte processes and corticostriatal neuronal projection terminals onto medium spiny neurons. We report the required protocols in detail, including adeno-associated virus microinjections, acute brain slice preparation, imaging, and post hoc FRET quantification. The article provides a detailed description that can be used to characterize and study astrocyte process proximity to synapses in living tissue. © 2020 by John Wiley & Sons, Inc. Basic Protocol 1: Forster resonance energy transfer imaging in cultured cells Basic Protocol 2: Forster resonance energy transfer imaging with the neuron-astrocyte proximity assay in acute brain slices.