A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials
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...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435330/ https://www.ncbi.nlm.nih.gov/pubmed/22970127 http://dx.doi.org/10.1371/journal.pone.0043454 |
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author | Barnett, Lauren Platisa, Jelena Popovic, Marko Pieribone, Vincent A. Hughes, Thomas |
author_facet | Barnett, Lauren Platisa, Jelena Popovic, Marko Pieribone, Vincent A. Hughes, Thomas |
author_sort | Barnett, Lauren |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3435330 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-34353302012-09-11 A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials Barnett, Lauren Platisa, Jelena Popovic, Marko Pieribone, Vincent A. Hughes, Thomas PLoS One Research Article 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. Public Library of Science 2012-09-06 /pmc/articles/PMC3435330/ /pubmed/22970127 http://dx.doi.org/10.1371/journal.pone.0043454 Text en © 2012 Barnett et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Barnett, Lauren Platisa, Jelena Popovic, Marko Pieribone, Vincent A. Hughes, Thomas A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials |
title | A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials |
title_full | A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials |
title_fullStr | A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials |
title_full_unstemmed | A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials |
title_short | A Fluorescent, Genetically-Encoded Voltage Probe Capable of Resolving Action Potentials |
title_sort | fluorescent, genetically-encoded voltage probe capable of resolving action potentials |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3435330/ https://www.ncbi.nlm.nih.gov/pubmed/22970127 http://dx.doi.org/10.1371/journal.pone.0043454 |
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