Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor

Víctor Fernández-Dueñas; Xavier Morató; Thomas Knöpfel; Francisco Ciruela

doi:10.1007/978-1-4939-3064-7_27

Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor

Víctor Fernández-Dueñas^*
, Xavier Morató
, Thomas Knöpfel
, Francisco Ciruela

^*Corresponding author for this work

Department of Physics

Research output: Chapter in book/report/conference proceeding › Chapter › peer-review

1 Citation (Scopus)

Abstract

Fluorescence-based biosensors for membrane voltage (mV) allow dynamic optical recording of neuronal activity. Interestingly, the development of genetically encoded voltage indicators constitute a good alternative to classical voltage-sensitive dyes, thus allowing overcoming some of the inherent problems (e.g., optical noise, etc.) associated with these organic compounds. Here, we show the use of a genetically encoded voltage-sensitive fluorescent protein (VSFP), namely the VSFP2.32, which contains a mCerulean and Citrine tandem engaging in a constitutive fluorescent resonance energy transfer (FRET) process. By expressing VSFP2.32 in hippocampal cultured neurons, we were able to monitor mV alterations in single neurons by recording VSFP2.32 conformation-mediated FRET changes in a real-time mode.

Original language	English
Title of host publication	Receptor and Ion Channel Detection in the Brain
Subtitle of host publication	Methods and Protocols
Editors	Rafael Luján, Francisco Ciruela
Publisher	Humana New York, NY
Chapter	27
Pages	447-454
Number of pages	8
Edition	1st
ISBN (Electronic)	9781493930647
ISBN (Print)	9781493930630, 9781493979837
DOIs	https://doi.org/10.1007/978-1-4939-3064-7_27
Publication status	Published - 2 Feb 2016

Publication series

Name	Neuromethods
Publisher	Humana Press
Volume	110
ISSN (Print)	0893-2336
ISSN (Electronic)	1940-6045

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

User-Defined Keywords

FRET
Neuronal membrane potential
Voltage indicators

Access to Document

10.1007/978-1-4939-3064-7_27

Cite this

Fernández-Dueñas, V., Morató, X., Knöpfel, T., & Ciruela, F. (2016). Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor. In R. Luján, & F. Ciruela (Eds.), Receptor and Ion Channel Detection in the Brain: Methods and Protocols (1st ed., pp. 447-454). (Neuromethods; Vol. 110). Humana New York, NY. https://doi.org/10.1007/978-1-4939-3064-7_27

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abstract = "Fluorescence-based biosensors for membrane voltage (mV) allow dynamic optical recording of neuronal activity. Interestingly, the development of genetically encoded voltage indicators constitute a good alternative to classical voltage-sensitive dyes, thus allowing overcoming some of the inherent problems (e.g., optical noise, etc.) associated with these organic compounds. Here, we show the use of a genetically encoded voltage-sensitive fluorescent protein (VSFP), namely the VSFP2.32, which contains a mCerulean and Citrine tandem engaging in a constitutive fluorescent resonance energy transfer (FRET) process. By expressing VSFP2.32 in hippocampal cultured neurons, we were able to monitor mV alterations in single neurons by recording VSFP2.32 conformation-mediated FRET changes in a real-time mode.",

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author = "V{\'i}ctor Fern{\'a}ndez-Due{\~n}as and Xavier Morat{\'o} and Thomas Kn{\"o}pfel and Francisco Ciruela",

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Fernández-Dueñas, V, Morató, X, Knöpfel, T & Ciruela, F 2016, Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor. in R Luján & F Ciruela (eds), Receptor and Ion Channel Detection in the Brain: Methods and Protocols. 1st edn, Neuromethods, vol. 110, Humana New York, NY, pp. 447-454. https://doi.org/10.1007/978-1-4939-3064-7_27

Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor. / Fernández-Dueñas, Víctor; Morató, Xavier; Knöpfel, Thomas et al.
Receptor and Ion Channel Detection in the Brain: Methods and Protocols. ed. / Rafael Luján; Francisco Ciruela. 1st. ed. Humana New York, NY, 2016. p. 447-454 (Neuromethods; Vol. 110).

Research output: Chapter in book/report/conference proceeding › Chapter › peer-review

TY - CHAP

T1 - Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor

AU - Fernández-Dueñas, Víctor

AU - Morató, Xavier

AU - Knöpfel, Thomas

AU - Ciruela, Francisco

PY - 2016/2/2

Y1 - 2016/2/2

N2 - Fluorescence-based biosensors for membrane voltage (mV) allow dynamic optical recording of neuronal activity. Interestingly, the development of genetically encoded voltage indicators constitute a good alternative to classical voltage-sensitive dyes, thus allowing overcoming some of the inherent problems (e.g., optical noise, etc.) associated with these organic compounds. Here, we show the use of a genetically encoded voltage-sensitive fluorescent protein (VSFP), namely the VSFP2.32, which contains a mCerulean and Citrine tandem engaging in a constitutive fluorescent resonance energy transfer (FRET) process. By expressing VSFP2.32 in hippocampal cultured neurons, we were able to monitor mV alterations in single neurons by recording VSFP2.32 conformation-mediated FRET changes in a real-time mode.

AB - Fluorescence-based biosensors for membrane voltage (mV) allow dynamic optical recording of neuronal activity. Interestingly, the development of genetically encoded voltage indicators constitute a good alternative to classical voltage-sensitive dyes, thus allowing overcoming some of the inherent problems (e.g., optical noise, etc.) associated with these organic compounds. Here, we show the use of a genetically encoded voltage-sensitive fluorescent protein (VSFP), namely the VSFP2.32, which contains a mCerulean and Citrine tandem engaging in a constitutive fluorescent resonance energy transfer (FRET) process. By expressing VSFP2.32 in hippocampal cultured neurons, we were able to monitor mV alterations in single neurons by recording VSFP2.32 conformation-mediated FRET changes in a real-time mode.

KW - FRET

KW - Neuronal membrane potential

KW - Voltage indicators

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SN - 9781493979837

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A2 - Luján, Rafael

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PB - Humana New York, NY

ER -

Dynamic recording of membrane potential from hippocampal neurons by using a FRET-based voltage biosensor

Abstract

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