High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein

Giuseppe Maulucci; Valentina Labate; Marina Mele; Emiliano Panieri; Giuseppe Arcovito; Tommaso Galeotti; Henrik Østergaard; Jakob R Winther; Marco De Spirito; Giovambattista Pani

doi:10.1126/scisignal.143pl3

High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein

Giuseppe Maulucci, Valentina Labate, Marina Mele, Emiliano Panieri, Giuseppe Arcovito, Tommaso Galeotti, Henrik Østergaard, Jakob R Winther, Marco De Spirito, Giovambattista Pani

Biomolecular Sciences

43 Citations (Scopus)

Abstract

We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time "redox maps" of adherent cells and to score subtle changes in the intracellular redox state, such as those induced by overexpression of redox-active proteins. This strategy for in vivo imaging of redox signaling circumvents many of the technical limitations currently encountered in the study of complex redox-based phenomena and promises to contribute substantially to this expanding area of signal transduction.

Original language	English
Journal	Science Signaling
Volume	1
Issue number	43
Pages (from-to)	pl3 1-15
Number of pages	15
DOIs	https://doi.org/10.1126/scisignal.143pl3
Publication status	Published - 2008

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title = "High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein",

abstract = "We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time {"}redox maps{"} of adherent cells and to score subtle changes in the intracellular redox state, such as those induced by overexpression of redox-active proteins. This strategy for in vivo imaging of redox signaling circumvents many of the technical limitations currently encountered in the study of complex redox-based phenomena and promises to contribute substantially to this expanding area of signal transduction.",

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T1 - High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein

AU - Maulucci, Giuseppe

AU - Labate, Valentina

AU - Mele, Marina

AU - Panieri, Emiliano

AU - Arcovito, Giuseppe

AU - Galeotti, Tommaso

AU - Østergaard, Henrik

AU - Winther, Jakob R

AU - De Spirito, Marco

AU - Pani, Giovambattista

PY - 2008

Y1 - 2008

N2 - We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time "redox maps" of adherent cells and to score subtle changes in the intracellular redox state, such as those induced by overexpression of redox-active proteins. This strategy for in vivo imaging of redox signaling circumvents many of the technical limitations currently encountered in the study of complex redox-based phenomena and promises to contribute substantially to this expanding area of signal transduction.

AB - We present the application of a redox-sensitive mutant of the yellow fluorescent protein (rxYFP) to image, with elevated sensitivity and high temporal and spatial resolution, oxidative responses of eukaryotic cells to pathophysiological stimuli. The method presented, based on the ratiometric quantitation of the distribution of fluorescence by confocal microscopy, allows us to draw real-time "redox maps" of adherent cells and to score subtle changes in the intracellular redox state, such as those induced by overexpression of redox-active proteins. This strategy for in vivo imaging of redox signaling circumvents many of the technical limitations currently encountered in the study of complex redox-based phenomena and promises to contribute substantially to this expanding area of signal transduction.

U2 - 10.1126/scisignal.143pl3

DO - 10.1126/scisignal.143pl3

M3 - Journal article

C2 - 18957692

SN - 1937-9145

VL - 1

SP - pl3 1-15

JO - Science Signaling

JF - Science Signaling

IS - 43

ER -

High-resolution imaging of redox signaling in live cells through an oxidation-sensitive yellow fluorescent protein

Abstract

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