Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.

Cyclic GMP (cGMP) regulates many physiological processes by cooperating with the other signaling molecules such as cyclic AMP (cAMP) and Ca(2+). Genetically encoded sensors for cGMP have been developed based on fluorescence resonance energy transfer (FRET) between fluorescent proteins. However, to a...

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Autores principales:	Yusuke Niino, Kohji Hotta, Kotaro Oka
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8468c4daa9754fee8337611e2f7df7c4
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spelling	oai:doaj.org-article:8468c4daa9754fee8337611e2f7df7c42021-11-25T06:25:55ZBlue fluorescent cGMP sensor for multiparameter fluorescence imaging.1932-620310.1371/journal.pone.0009164https://doaj.org/article/8468c4daa9754fee8337611e2f7df7c42010-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20161796/?tool=EBIhttps://doaj.org/toc/1932-6203Cyclic GMP (cGMP) regulates many physiological processes by cooperating with the other signaling molecules such as cyclic AMP (cAMP) and Ca(2+). Genetically encoded sensors for cGMP have been developed based on fluorescence resonance energy transfer (FRET) between fluorescent proteins. However, to analyze the dynamic relationship among these second messengers, combined use of existing sensors in a single cell is inadequate because of the significant spectral overlaps. A single wavelength indicator is an effective alternative to avoid this problem, but color variants of a single fluorescent protein-based biosensor are limited. In this study, to construct a new color fluorescent sensor, we converted the FRET-based sensor into a single wavelength indicator using a dark FRET acceptor. We developed a blue fluorescent cGMP biosensor, which is spectrally compatible with a FRET-based cAMP sensor using cyan and yellow fluorescent proteins (CFP/YFP). We cotransfected them and loaded a red fluorescent probe for Ca(2+) into cells, and accomplished triple-parameter fluorescence imaging of these cyclic nucleotides and Ca(2+), confirming the applicability of this combination to individually monitor their dynamics in a single cell. This blue fluorescent sensor and the approach using this FRET pair would be useful for multiparameter fluorescence imaging to understand complex signal transduction networks.Yusuke NiinoKohji HottaKotaro OkaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 2, p e9164 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yusuke Niino Kohji Hotta Kotaro Oka Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
description	Cyclic GMP (cGMP) regulates many physiological processes by cooperating with the other signaling molecules such as cyclic AMP (cAMP) and Ca(2+). Genetically encoded sensors for cGMP have been developed based on fluorescence resonance energy transfer (FRET) between fluorescent proteins. However, to analyze the dynamic relationship among these second messengers, combined use of existing sensors in a single cell is inadequate because of the significant spectral overlaps. A single wavelength indicator is an effective alternative to avoid this problem, but color variants of a single fluorescent protein-based biosensor are limited. In this study, to construct a new color fluorescent sensor, we converted the FRET-based sensor into a single wavelength indicator using a dark FRET acceptor. We developed a blue fluorescent cGMP biosensor, which is spectrally compatible with a FRET-based cAMP sensor using cyan and yellow fluorescent proteins (CFP/YFP). We cotransfected them and loaded a red fluorescent probe for Ca(2+) into cells, and accomplished triple-parameter fluorescence imaging of these cyclic nucleotides and Ca(2+), confirming the applicability of this combination to individually monitor their dynamics in a single cell. This blue fluorescent sensor and the approach using this FRET pair would be useful for multiparameter fluorescence imaging to understand complex signal transduction networks.
format	article
author	Yusuke Niino Kohji Hotta Kotaro Oka
author_facet	Yusuke Niino Kohji Hotta Kotaro Oka
author_sort	Yusuke Niino
title	Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
title_short	Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
title_full	Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
title_fullStr	Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
title_full_unstemmed	Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.
title_sort	blue fluorescent cgmp sensor for multiparameter fluorescence imaging.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/8468c4daa9754fee8337611e2f7df7c4
work_keys_str_mv	AT yusukeniino bluefluorescentcgmpsensorformultiparameterfluorescenceimaging AT kohjihotta bluefluorescentcgmpsensorformultiparameterfluorescenceimaging AT kotarooka bluefluorescentcgmpsensorformultiparameterfluorescenceimaging
_version_	1718413755594833920

Blue fluorescent cGMP sensor for multiparameter fluorescence imaging.

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