Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging

Abstract cAMP is a common second messenger that is involved in various physiological processes. To expand the colour palette of available cAMP indicators, we developed a red cAMP indicator named “Pink Flamindo” (Pink Fluorescent cAMP indicator). The fluorescence intensity of Pink Flamindo increases...

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Autores principales:	Kazuki Harada, Motoki Ito, Xiaowen Wang, Mika Tanaka, Devina Wongso, Ayumu Konno, Hirokazu Hirai, Hajime Hirase, Takashi Tsuboi, Tetsuya Kitaguchi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a46cc053037049d8a7d8d5c8d75380a3
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spelling	oai:doaj.org-article:a46cc053037049d8a7d8d5c8d75380a32021-12-02T15:05:26ZRed fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging10.1038/s41598-017-07820-62045-2322https://doaj.org/article/a46cc053037049d8a7d8d5c8d75380a32017-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-07820-6https://doaj.org/toc/2045-2322Abstract cAMP is a common second messenger that is involved in various physiological processes. To expand the colour palette of available cAMP indicators, we developed a red cAMP indicator named “Pink Flamindo” (Pink Fluorescent cAMP indicator). The fluorescence intensity of Pink Flamindo increases 4.2-fold in the presence of a saturating dose of cAMP, with excitation and emission peaks at 567 nm and 590 nm, respectively. Live-cell imaging revealed that Pink Flamindo is effective for monitoring the spatio-temporal dynamics of intracellular cAMP generated by photoactivated adenylyl cyclase in response to blue light, and in dual-colour imaging studies using a green Ca2+ indicator (G-GECO). Furthermore, we successfully monitored the elevation of cAMP levels in vivo in cerebral cortical astrocytes by two-photon imaging. We propose that Pink Flamindo will facilitate future in vivo, optogenetic studies of cell signalling and cAMP dynamics.Kazuki HaradaMotoki ItoXiaowen WangMika TanakaDevina WongsoAyumu KonnoHirokazu HiraiHajime HiraseTakashi TsuboiTetsuya KitaguchiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-9 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kazuki Harada Motoki Ito Xiaowen Wang Mika Tanaka Devina Wongso Ayumu Konno Hirokazu Hirai Hajime Hirase Takashi Tsuboi Tetsuya Kitaguchi Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
description	Abstract cAMP is a common second messenger that is involved in various physiological processes. To expand the colour palette of available cAMP indicators, we developed a red cAMP indicator named “Pink Flamindo” (Pink Fluorescent cAMP indicator). The fluorescence intensity of Pink Flamindo increases 4.2-fold in the presence of a saturating dose of cAMP, with excitation and emission peaks at 567 nm and 590 nm, respectively. Live-cell imaging revealed that Pink Flamindo is effective for monitoring the spatio-temporal dynamics of intracellular cAMP generated by photoactivated adenylyl cyclase in response to blue light, and in dual-colour imaging studies using a green Ca2+ indicator (G-GECO). Furthermore, we successfully monitored the elevation of cAMP levels in vivo in cerebral cortical astrocytes by two-photon imaging. We propose that Pink Flamindo will facilitate future in vivo, optogenetic studies of cell signalling and cAMP dynamics.
format	article
author	Kazuki Harada Motoki Ito Xiaowen Wang Mika Tanaka Devina Wongso Ayumu Konno Hirokazu Hirai Hajime Hirase Takashi Tsuboi Tetsuya Kitaguchi
author_facet	Kazuki Harada Motoki Ito Xiaowen Wang Mika Tanaka Devina Wongso Ayumu Konno Hirokazu Hirai Hajime Hirase Takashi Tsuboi Tetsuya Kitaguchi
author_sort	Kazuki Harada
title	Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
title_short	Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
title_full	Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
title_fullStr	Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
title_full_unstemmed	Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
title_sort	red fluorescent protein-based camp indicator applicable to optogenetics and in vivo imaging
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/a46cc053037049d8a7d8d5c8d75380a3
work_keys_str_mv	AT kazukiharada redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT motokiito redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT xiaowenwang redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT mikatanaka redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT devinawongso redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT ayumukonno redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT hirokazuhirai redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT hajimehirase redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT takashitsuboi redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging AT tetsuyakitaguchi redfluorescentproteinbasedcampindicatorapplicabletooptogeneticsandinvivoimaging
_version_	1718388850628231168

Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging

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