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Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging
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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5544736/ https://www.ncbi.nlm.nih.gov/pubmed/28779099 http://dx.doi.org/10.1038/s41598-017-07820-6 |
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author | Harada, Kazuki Ito, Motoki Wang, Xiaowen Tanaka, Mika Wongso, Devina Konno, Ayumu Hirai, Hirokazu Hirase, Hajime Tsuboi, Takashi Kitaguchi, Tetsuya |
author_facet | Harada, Kazuki Ito, Motoki Wang, Xiaowen Tanaka, Mika Wongso, Devina Konno, Ayumu Hirai, Hirokazu Hirase, Hajime Tsuboi, Takashi Kitaguchi, Tetsuya |
author_sort | Harada, Kazuki |
collection | PubMed |
description | 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 Ca(2+) 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 | Online Article Text |
id | pubmed-5544736 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-55447362017-08-09 Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging Harada, Kazuki Ito, Motoki Wang, Xiaowen Tanaka, Mika Wongso, Devina Konno, Ayumu Hirai, Hirokazu Hirase, Hajime Tsuboi, Takashi Kitaguchi, Tetsuya Sci Rep Article 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 Ca(2+) 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. Nature Publishing Group UK 2017-08-04 /pmc/articles/PMC5544736/ /pubmed/28779099 http://dx.doi.org/10.1038/s41598-017-07820-6 Text en © The Author(s) 2017 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Harada, Kazuki Ito, Motoki Wang, Xiaowen Tanaka, Mika Wongso, Devina Konno, Ayumu Hirai, Hirokazu Hirase, Hajime Tsuboi, Takashi Kitaguchi, Tetsuya Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging |
title | 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_short | 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 |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5544736/ https://www.ncbi.nlm.nih.gov/pubmed/28779099 http://dx.doi.org/10.1038/s41598-017-07820-6 |
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