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Neuron. 2012 Sep 6;75(5):779-85. doi: 10.1016/j.neuron.2012.06.040.

Single action potentials and subthreshold electrical events imaged in neurons with a fluorescent protein voltage probe.

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1
Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06511, USA.

Abstract

Monitoring neuronal electrical activity using fluorescent protein-based voltage sensors has been limited by small response magnitudes and slow kinetics of existing probes. Here we report the development of a fluorescent protein voltage sensor, named ArcLight, and derivative probes that exhibit large changes in fluorescence intensity in response to voltage changes. ArcLight consists of the voltage-sensing domain of Ciona intestinalis voltage-sensitive phosphatase and super ecliptic pHluorin that carries the point mutation A227D. The fluorescence intensity of ArcLight A242 decreases by 35% in response to a 100 mV depolarization when measured in HEK293 cells, which is more than five times larger than the signals from previously reported fluorescent protein voltage sensors. We show that the combination of signal size and response speed of these new probes allows the reliable detection of single action potentials and excitatory potentials in individual neurons and dendrites.

PMID:
22958819
PMCID:
PMC3439164
DOI:
10.1016/j.neuron.2012.06.040
[Indexed for MEDLINE]
Free PMC Article
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