A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials(Article)(Open Access)
- aDepartment of Cell Biology and Neuroscience, Montana State University, Bozeman, MT, United States
- bThe John B. Pierce Laboratory, Inc., New Haven, CT, United States
- cFaculty of Physical Chemistry, University of Belgrade, Belgrade, Serbia
- dCellular and Molecular Physiology and Neurobiology, Yale University School of Medicine, New Haven, CT, United States
Abstract
There is a pressing need in neuroscience for genetically-encoded, fluorescent voltage probes that can be targeted to specific neurons and circuits to allow study of neural activity using fluorescent imaging. We created 90 constructs in which the voltage sensing portion (S1-S4) of Ciona intestinalis voltage sensitive phosphatase (CiVSP) was fused to circularly permuted eGFP. This led to ElectricPk, a probe that is an order of magnitude faster (taus ~1-2 ms) than any currently published fluorescent protein-based voltage probe. ElectricPk can follow the rise and fall of neuronal action potentials with a modest decrease in fluorescence intensity (~0.7% ΔF/F). The probe has a nearly linear fluorescence/membrane potential response to both hyperpolarizing and depolarizing steps. This is the first probe based on CiVSP that captures the rapid movements of the voltage sensor, suggesting that voltage probes designed with circularly permuted fluorescent proteins may have some advantages. © 2012 Barnett et al.
Indexed keywords
| EMTREE drug terms: | enhanced green fluorescent proteinphosphatase |
|---|---|
| EMTREE medical terms: | action potentialanimal cellarticleCiona intestinalisdepolarizationembryofluorescence imaginggeneral devicegenetically encoded fluorescent voltage probehumanhuman cellhyperpolarizationmolecular probemousenerve cellnonhumansensor |
| MeSH: | Action PotentialsAmino Acid SequenceAnimalsCiona intestinalisGenes, ReporterGreen Fluorescent ProteinsHEK293 CellsHippocampusHumansKineticsMiceMolecular Sequence DataNeuronsPhosphoric Monoester HydrolasesRecombinant Fusion Proteins |
| Species Index: | Ciona intestinalis |
Chemicals and CAS Registry Numbers:
phosphatase, 9013-05-2;
Green Fluorescent Proteins, 147336-22-9; Phosphoric Monoester Hydrolases, 3.1.3.-; Recombinant Fusion Proteins; enhanced green fluorescent protein; voltage-sensor-containing phosphatase, Ciona intestinalis, 3.1.3.-
Device tradename:
- ElectricPk
Funding details
| Funding sponsor | Funding number | Acronym |
|---|---|---|
| National Institute of Neurological Disorders and Stroke See opportunities by NINDS | U24NS057631 | NINDS |
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1
The authors have the following conflicts: Funding for this study came in part from The John B. Pierce Laboratory, Inc. Authors Jelena Platisa and Vincent A. Pieribone are employees of The John B. Pierce Laboratory, Inc. There are no other relevant declarations relating to employment, consultancy, patents, products in development or marketed products. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials.
- ISSN: 19326203
- Source Type: Journal
- Original language: English
- DOI: 10.1371/journal.pone.0043454
- PubMed ID: 22970127
- Document Type: Article
Pieribone, V. A.; The John B. Pierce Laboratory, Inc., United States;
© Copyright 2013 Elsevier B.V., All rights reserved.
© MEDLINE® is the source for the MeSH terms of this document.
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