Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition

Abstract An improved genetically encoded voltage indicator (GEVI) was achieved by altering the charge composition of the region linking the voltage-sensing domain of the GEVI to a pH-sensitive fluorescent protein. Negatively charged linker segments reduced the voltage-dependent optical signal while...

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Autores principales:	Sungmoo Lee, Tristan Geiller, Arong Jung, Ryuichi Nakajima, Yoon-Kyu Song, Bradley J. Baker
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fccba8cd2d084bddb5510817b58c16d4
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spelling	oai:doaj.org-article:fccba8cd2d084bddb5510817b58c16d42021-12-02T12:32:44ZImproving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition10.1038/s41598-017-08731-22045-2322https://doaj.org/article/fccba8cd2d084bddb5510817b58c16d42017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-08731-2https://doaj.org/toc/2045-2322Abstract An improved genetically encoded voltage indicator (GEVI) was achieved by altering the charge composition of the region linking the voltage-sensing domain of the GEVI to a pH-sensitive fluorescent protein. Negatively charged linker segments reduced the voltage-dependent optical signal while positively charged linkers increased the signal size. Arginine scanning mutagenesis of the linker region improved the signal size of the GEVI, Bongwoori, yielding fluorescent signals as high as 20% ΔF/F during the firing of action potentials. The speed of this new sensor was also capable of optically resolving action potentials firing at 65 Hz. This large signal size enabled individual pixels to become surrogate electrodes. Plotting the highest correlated pixels based only on fluorescence changes reproduced the image of the neuron exhibiting activity. Furthermore, the use of a pH-sensitive fluorescent protein facilitated the detection of the acidification of the neuron during the firing of action potentials.Sungmoo LeeTristan GeillerArong JungRyuichi NakajimaYoon-Kyu SongBradley J. BakerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-16 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Sungmoo Lee Tristan Geiller Arong Jung Ryuichi Nakajima Yoon-Kyu Song Bradley J. Baker Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
description	Abstract An improved genetically encoded voltage indicator (GEVI) was achieved by altering the charge composition of the region linking the voltage-sensing domain of the GEVI to a pH-sensitive fluorescent protein. Negatively charged linker segments reduced the voltage-dependent optical signal while positively charged linkers increased the signal size. Arginine scanning mutagenesis of the linker region improved the signal size of the GEVI, Bongwoori, yielding fluorescent signals as high as 20% ΔF/F during the firing of action potentials. The speed of this new sensor was also capable of optically resolving action potentials firing at 65 Hz. This large signal size enabled individual pixels to become surrogate electrodes. Plotting the highest correlated pixels based only on fluorescence changes reproduced the image of the neuron exhibiting activity. Furthermore, the use of a pH-sensitive fluorescent protein facilitated the detection of the acidification of the neuron during the firing of action potentials.
format	article
author	Sungmoo Lee Tristan Geiller Arong Jung Ryuichi Nakajima Yoon-Kyu Song Bradley J. Baker
author_facet	Sungmoo Lee Tristan Geiller Arong Jung Ryuichi Nakajima Yoon-Kyu Song Bradley J. Baker
author_sort	Sungmoo Lee
title	Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
title_short	Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
title_full	Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
title_fullStr	Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
title_full_unstemmed	Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
title_sort	improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/fccba8cd2d084bddb5510817b58c16d4
work_keys_str_mv	AT sungmoolee improvingageneticallyencodedvoltageindicatorbymodifyingthecytoplasmicchargecomposition AT tristangeiller improvingageneticallyencodedvoltageindicatorbymodifyingthecytoplasmicchargecomposition AT arongjung improvingageneticallyencodedvoltageindicatorbymodifyingthecytoplasmicchargecomposition AT ryuichinakajima improvingageneticallyencodedvoltageindicatorbymodifyingthecytoplasmicchargecomposition AT yoonkyusong improvingageneticallyencodedvoltageindicatorbymodifyingthecytoplasmicchargecomposition AT bradleyjbaker improvingageneticallyencodedvoltageindicatorbymodifyingthecytoplasmicchargecomposition
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Improving a genetically encoded voltage indicator by modifying the cytoplasmic charge composition

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