Imaging neural spiking in brain tissue using FRET-opsin protein voltage sensors

Yiyang Gong (), Mark J. Wagner, Jin Zhong Li and Mark J. Schnitzer ()
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Yiyang Gong: James H. Clark Center, Stanford University
Mark J. Wagner: James H. Clark Center, Stanford University
Jin Zhong Li: James H. Clark Center, Stanford University
Mark J. Schnitzer: James H. Clark Center, Stanford University

Nature Communications, 2014, vol. 5, issue 1, 1-11

Abstract: Abstract Genetically encoded fluorescence voltage sensors offer the possibility of directly visualizing neural spiking dynamics in cells targeted by their genetic class or connectivity. Sensors of this class have generally suffered performance-limiting tradeoffs between modest brightness, sluggish kinetics and limited signalling dynamic range in response to action potentials. Here we describe sensors that use fluorescence resonance energy transfer (FRET) to combine the rapid kinetics and substantial voltage-dependence of rhodopsin family voltage-sensing domains with the brightness of genetically engineered protein fluorophores. These FRET-opsin sensors significantly improve upon the spike detection fidelity offered by the genetically encoded voltage sensor, Arclight, while offering faster kinetics and higher brightness. Using FRET-opsin sensors we imaged neural spiking and sub-threshold membrane voltage dynamics in cultured neurons and in pyramidal cells within neocortical tissue slices. In live mice, rates and optical waveforms of cerebellar Purkinje neurons’ dendritic voltage transients matched expectations for these cells’ dendritic spikes.

Date: 2014
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DOI: 10.1038/ncomms4674

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