Imaging synaptic inhibition with the genetically encoded chloride indicator Clomeleon.
Imaging synaptic inhibition with the genetically encoded chloride indicator Clomeleon.
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DOI：
10.1101/pdb.prot066985
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发表时间：
2011-12-01
影响因子：
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通讯作者：
Augustine, George J
中科院分区：
文献类型：
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作者：
Berglund, Ken;Kuner, Thomas;Augustine, George J
有几种技术可以对大脑中的兴奋过程进行成像，但突触抑制在很大程度上仍然是不可见的。大脑中的大多数突触抑制源于氯离子 (Cl(-)) 的跨膜通量，因此细胞内 Cl(-) 浓度成像 ([Cl(-)](i)) 原则上是可视化抑制时空动态的自然方法。该协议描述了 Clomeleon（一种 Cl(-) 的基因编码指示剂）的使用，作为监测突触抑制的工具。它概述了可用于对 Clomeleon 转基因小鼠制备的脑切片中的神经元 [Cl(-)](i) 进行成像的程序。只需进行微小的调整，相同的程序也应该适用于培养细胞的成像。
Several techniques are available to image excitatory processes in the brain, but synaptic inhibition has remained largely invisible. Most synaptic inhibition in the brain arises from transmembrane fluxes of chloride ions (Cl(-)), so imaging intracellular Cl(-) concentration ([Cl(-)](i)) is, in principle, a natural way to visualize the spatiotemporal dynamics of inhibition. This protocol describes the use of Clomeleon, a genetically encoded indicator of Cl(-), as a tool for monitoring synaptic inhibition. It outlines procedures that can be used to image neuronal [Cl(-)](i) in brain slices prepared from Clomeleon transgenic mice. With only minor adjustments, the same procedures should be suitable for imaging from cultured cells as well.