Imaging Synaptic Inhibition with the Genetically Encoded Chloride Indicator Clomeleon

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Abstract
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.