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Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags

The stochastic multicolor labeling method ‘Brainbow’ is a powerful strategy to label multiple neurons differentially with fluorescent proteins; however, the fluorescence levels provided by the original attempts to use this strategy were inadequate. In the present study, we developed a stochastic mul...

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Detalles Bibliográficos
Autores principales: Sakaguchi, Richi, Leiwe, Marcus N, Imai, Takeshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6245733/
https://www.ncbi.nlm.nih.gov/pubmed/30454553
http://dx.doi.org/10.7554/eLife.40350
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author Sakaguchi, Richi
Leiwe, Marcus N
Imai, Takeshi
author_facet Sakaguchi, Richi
Leiwe, Marcus N
Imai, Takeshi
author_sort Sakaguchi, Richi
collection PubMed
description The stochastic multicolor labeling method ‘Brainbow’ is a powerful strategy to label multiple neurons differentially with fluorescent proteins; however, the fluorescence levels provided by the original attempts to use this strategy were inadequate. In the present study, we developed a stochastic multicolor labeling method with enhanced expression levels that uses a tetracycline-operator system (Tetbow). We optimized Tetbow for either plasmid or virus vector-mediated multicolor labeling. When combined with tissue clearing, Tetbow was powerful enough to visualize the three-dimensional architecture of individual neurons. Using Tetbow, we were able to visualize the axonal projection patterns of individual mitral/tufted cells along several millimeters in the mouse olfactory system. We also developed a Tetbow system with chemical tags, in which genetically encoded chemical tags were labeled with synthetic fluorophores. This was useful in expanding the repertoire of the fluorescence labels and the applications of the Tetbow system. Together, these new tools facilitate light-microscopy-based neuronal tracing at both a large scale and a high resolution.
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spelling pubmed-62457332018-11-20 Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags Sakaguchi, Richi Leiwe, Marcus N Imai, Takeshi eLife Neuroscience The stochastic multicolor labeling method ‘Brainbow’ is a powerful strategy to label multiple neurons differentially with fluorescent proteins; however, the fluorescence levels provided by the original attempts to use this strategy were inadequate. In the present study, we developed a stochastic multicolor labeling method with enhanced expression levels that uses a tetracycline-operator system (Tetbow). We optimized Tetbow for either plasmid or virus vector-mediated multicolor labeling. When combined with tissue clearing, Tetbow was powerful enough to visualize the three-dimensional architecture of individual neurons. Using Tetbow, we were able to visualize the axonal projection patterns of individual mitral/tufted cells along several millimeters in the mouse olfactory system. We also developed a Tetbow system with chemical tags, in which genetically encoded chemical tags were labeled with synthetic fluorophores. This was useful in expanding the repertoire of the fluorescence labels and the applications of the Tetbow system. Together, these new tools facilitate light-microscopy-based neuronal tracing at both a large scale and a high resolution. eLife Sciences Publications, Ltd 2018-11-20 /pmc/articles/PMC6245733/ /pubmed/30454553 http://dx.doi.org/10.7554/eLife.40350 Text en © 2018, Sakaguchi et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Neuroscience
Sakaguchi, Richi
Leiwe, Marcus N
Imai, Takeshi
Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
title Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
title_full Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
title_fullStr Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
title_full_unstemmed Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
title_short Bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
title_sort bright multicolor labeling of neuronal circuits with fluorescent proteins and chemical tags
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6245733/
https://www.ncbi.nlm.nih.gov/pubmed/30454553
http://dx.doi.org/10.7554/eLife.40350
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