Neuron
Volume 27, Issue 3, September 2000, Pages 447-459
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Neurotechnique
A Genetically Encoded Ratiometric Indicator for Chloride: Capturing Chloride Transients in Cultured Hippocampal Neurons

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Abstract

We constructed a novel optical indicator for chloride ions by fusing the chloride-sensitive yellow fluorescent protein with the chloride-insensitive cyan fluorescent protein. The ratio of FRET-dependent emission of these fluorophores varied in proportion to the concentration of Cl and was used to measure intracellular chloride concentration ([Cl]i) in cultured hippocampal neurons. [Cl]i decreased during neuronal development, consistent with the shift from excitation to inhibition during maturation of GABAergic synapses. Focal activation of GABAA receptors caused large changes in [Cl]i that could underlie use-dependent depression of GABA-dependent synaptic transmission. GABA-induced changes in somatic [Cl]i spread into dendrites, suggesting that [Cl]i can signal the location of synaptic activity. This genetically encoded indicator will permit new approaches ranging from high-throughput drug screening to direct recordings of synaptic Cl signals in vivo.

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