Using Electrophysiology to Study Synaptic and Extrasynaptic Ionotropic Receptors in Hippocampal Neurons
Using Electrophysiology to Study Synaptic and Extrasynaptic Ionotropic Receptors in Hippocampal Neurons
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DOI：
10.1007/978-1-4939-3064-7_21
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发表时间：
2016-01-01
期刊：
影响因子：
--
通讯作者：
Soto, David
中科院分区：
文献类型：
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作者：
Coombs, Ian D.;Soto, David
电生理学是神经科学研究的一种特别有用的工具，因为神经元固有的电兴奋性以及神经元和神经胶质细胞中存在的大量离子通道。不同的电生理学技术可以应用于神经生物学，从测量广阔脑区的细胞群到测量质膜斑块中的单通道。电生理学作为一种工具的优势之一是能够测量异种系统中已知离子通道的特性，然后剖析神经元反应的各种药理学和生物物理学，最终更好地了解哪些成分通道及其特征决定了神经元和电路的生物关键输出。膜片钳电生理学允许记录突触和突触外水平的神经元受体。这里描述的特定技术允许通过测量微小的兴奋性突触电流和来自体细胞受体的电流来独立地研究这两个群体。本章所述技术的分辨率和精确度都很高(在亚皮安和亚毫秒范围内)。此外，这些方法提供了关于受体在其与辅助、调节和锚定蛋白共存的自然环境中的行为的有价值的信息。在本章中，我们首先描述了制备海马神经元培养的方法。其次，描述了记录和分析微小突触后电流的过程。最后，我们详细描述了将快速激动剂应用于从神经元胞体获得的外向贴片的技术。我们讨论了这些方法发现的常见问题，并提出了一些提示，以帮助新进入该领域的研究人员，以便他们可以迅速掌握这些技术。
Electrophysiology is an exceptionally useful tool for neuroscience research due to the intrinsic electrical excitability of neurons and the significant array of ion channels present in both neurons and glia. Diverse electrophysiological techniques may be applied to neurobiology ranging from measurements of cell populations in broad brain regions to measurements of single channels in patches of plasma membrane. One of the strengths of electrophysiology as a tool is the ability to measure the properties of known ion channels in heterologous systems, then dissect the diverse pharmacology and biophysics of neuronal responses to finally better understand which component channels and their features determine the biologically critical outputs of neurons and circuits.Patch-clamp electrophysiology allows recording of neuronal receptors at both the synaptic and extra-synaptic level. The specific techniques described here permit the study of both populations independently by measuring miniature excitatory synaptic currents and currents derived from somatic receptors. The resolution and accuracy of the techniques described in the chapter are high (in the sub-picoampere and sub-millisecond ranges). Further, these methodologies provide valuable information about the behavior of the receptors in their native environment where they coexist with auxiliary, modulatory, and anchoring proteins.In this chapter, we firstly describe the methodology for preparing hippocampal neuronal cultures. Secondly, we describe the process of recording and analyzing miniature postsynaptic currents. Finally, we describe in detail the technique of fast agonist application onto outside-out patches obtained from the soma of neurons. We discuss common problems found with these approaches and present tips to assist researchers new to the field so they may rapidly master the techniques.