Neuron
Volume 75, Issue 5, 6 September 2012, Pages 779-785
Journal home page for Neuron

NeuroResource
Single Action Potentials and Subthreshold Electrical Events Imaged in Neurons with a Fluorescent Protein Voltage Probe

https://doi.org/10.1016/j.neuron.2012.06.040Get rights and content
Under an Elsevier user license
open archive

Summary

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 100mV 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.

Highlights

► CiVS-based FP voltage sensors improved by A227D mutation in super ecliptic pHluorin ► Optogenetic visualization of single action potentials and subthreshold events

Cited by (0)

7

These authors contributed equally to this work