Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.

Imaging the activities of individual neurons with genetically encoded Ca(2+) indicators (GECIs) is a promising method for understanding neuronal network functions. Here, we report GECIs with improved neuronal Ca(2+) signal detectability, termed G-CaMP6 and G-CaMP8. Compared to a series of existing G...

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Autores principales:	Masamichi Ohkura, Takuya Sasaki, Junko Sadakari, Keiko Gengyo-Ando, Yuko Kagawa-Nagamura, Chiaki Kobayashi, Yuji Ikegaya, Junichi Nakai
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/26c1cd1f8311490492d0b62125fa491d
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spelling	oai:doaj.org-article:26c1cd1f8311490492d0b62125fa491d2021-11-18T08:05:36ZGenetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.1932-620310.1371/journal.pone.0051286https://doaj.org/article/26c1cd1f8311490492d0b62125fa491d2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23240011/?tool=EBIhttps://doaj.org/toc/1932-6203Imaging the activities of individual neurons with genetically encoded Ca(2+) indicators (GECIs) is a promising method for understanding neuronal network functions. Here, we report GECIs with improved neuronal Ca(2+) signal detectability, termed G-CaMP6 and G-CaMP8. Compared to a series of existing G-CaMPs, G-CaMP6 showed fairly high sensitivity and rapid kinetics, both of which are suitable properties for detecting subtle and fast neuronal activities. G-CaMP8 showed a greater signal (F(max)/F(min) = 38) than G-CaMP6 and demonstrated kinetics similar to those of G-CaMP6. Both GECIs could detect individual spikes from pyramidal neurons of cultured hippocampal slices or acute cortical slices with 100% detection rates, demonstrating their superior performance to existing GECIs. Because G-CaMP6 showed a higher sensitivity and brighter baseline fluorescence than G-CaMP8 in a cellular environment, we applied G-CaMP6 for Ca(2+) imaging of dendritic spines, the putative postsynaptic sites. By expressing a G-CaMP6-actin fusion protein for the spines in hippocampal CA3 pyramidal neurons and electrically stimulating the granule cells of the dentate gyrus, which innervate CA3 pyramidal neurons, we found that sub-threshold stimulation triggered small Ca(2+) responses in a limited number of spines with a low response rate in active spines, whereas supra-threshold stimulation triggered large fluorescence responses in virtually all of the spines with a 100% activity rate.Masamichi OhkuraTakuya SasakiJunko SadakariKeiko Gengyo-AndoYuko Kagawa-NagamuraChiaki KobayashiYuji IkegayaJunichi NakaiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e51286 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Masamichi Ohkura Takuya Sasaki Junko Sadakari Keiko Gengyo-Ando Yuko Kagawa-Nagamura Chiaki Kobayashi Yuji Ikegaya Junichi Nakai Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.
description	Imaging the activities of individual neurons with genetically encoded Ca(2+) indicators (GECIs) is a promising method for understanding neuronal network functions. Here, we report GECIs with improved neuronal Ca(2+) signal detectability, termed G-CaMP6 and G-CaMP8. Compared to a series of existing G-CaMPs, G-CaMP6 showed fairly high sensitivity and rapid kinetics, both of which are suitable properties for detecting subtle and fast neuronal activities. G-CaMP8 showed a greater signal (F(max)/F(min) = 38) than G-CaMP6 and demonstrated kinetics similar to those of G-CaMP6. Both GECIs could detect individual spikes from pyramidal neurons of cultured hippocampal slices or acute cortical slices with 100% detection rates, demonstrating their superior performance to existing GECIs. Because G-CaMP6 showed a higher sensitivity and brighter baseline fluorescence than G-CaMP8 in a cellular environment, we applied G-CaMP6 for Ca(2+) imaging of dendritic spines, the putative postsynaptic sites. By expressing a G-CaMP6-actin fusion protein for the spines in hippocampal CA3 pyramidal neurons and electrically stimulating the granule cells of the dentate gyrus, which innervate CA3 pyramidal neurons, we found that sub-threshold stimulation triggered small Ca(2+) responses in a limited number of spines with a low response rate in active spines, whereas supra-threshold stimulation triggered large fluorescence responses in virtually all of the spines with a 100% activity rate.
format	article
author	Masamichi Ohkura Takuya Sasaki Junko Sadakari Keiko Gengyo-Ando Yuko Kagawa-Nagamura Chiaki Kobayashi Yuji Ikegaya Junichi Nakai
author_facet	Masamichi Ohkura Takuya Sasaki Junko Sadakari Keiko Gengyo-Ando Yuko Kagawa-Nagamura Chiaki Kobayashi Yuji Ikegaya Junichi Nakai
author_sort	Masamichi Ohkura
title	Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.
title_short	Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.
title_full	Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.
title_fullStr	Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.
title_full_unstemmed	Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.
title_sort	genetically encoded green fluorescent ca2+ indicators with improved detectability for neuronal ca2+ signals.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/26c1cd1f8311490492d0b62125fa491d
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Genetically encoded green fluorescent Ca2+ indicators with improved detectability for neuronal Ca2+ signals.

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