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Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting
Circadian clocks are self-sustained and cell-autonomous oscillators. They respond to various extracellular cues depending on the time-of-day and the signal intensity. Phase Transition Curves (PTCs) are instrumental in uncovering the full repertoire of responses to a given signal. However, the curren...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8501123/ https://www.ncbi.nlm.nih.gov/pubmed/34625543 http://dx.doi.org/10.1038/s41467-021-26210-1 |
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author | Manella, Gal Aizik, Dan Aviram, Rona Golik, Marina Asher, Gad |
author_facet | Manella, Gal Aizik, Dan Aviram, Rona Golik, Marina Asher, Gad |
author_sort | Manella, Gal |
collection | PubMed |
description | Circadian clocks are self-sustained and cell-autonomous oscillators. They respond to various extracellular cues depending on the time-of-day and the signal intensity. Phase Transition Curves (PTCs) are instrumental in uncovering the full repertoire of responses to a given signal. However, the current methodologies for reconstructing PTCs are low-throughput, laborious, and resource- and time-consuming. We report here the development of an efficient and high throughput assay, dubbed Circadian Single-Cell Oscillators PTC Extraction (Circa-SCOPE) for generating high-resolution PTCs. This methodology relies on continuous monitoring of single-cell oscillations to reconstruct a full PTC from a single culture, upon a one-time intervention. Using Circa-SCOPE, we characterize the effects of various pharmacological and blood-borne resetting cues, at high temporal resolution and a wide concentration range. Thus, Circa-SCOPE is a powerful tool for comprehensive analysis and screening for circadian clocks’ resetting cues, and can be valuable for basic as well as translational research. |
format | Online Article Text |
id | pubmed-8501123 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-85011232021-10-22 Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting Manella, Gal Aizik, Dan Aviram, Rona Golik, Marina Asher, Gad Nat Commun Article Circadian clocks are self-sustained and cell-autonomous oscillators. They respond to various extracellular cues depending on the time-of-day and the signal intensity. Phase Transition Curves (PTCs) are instrumental in uncovering the full repertoire of responses to a given signal. However, the current methodologies for reconstructing PTCs are low-throughput, laborious, and resource- and time-consuming. We report here the development of an efficient and high throughput assay, dubbed Circadian Single-Cell Oscillators PTC Extraction (Circa-SCOPE) for generating high-resolution PTCs. This methodology relies on continuous monitoring of single-cell oscillations to reconstruct a full PTC from a single culture, upon a one-time intervention. Using Circa-SCOPE, we characterize the effects of various pharmacological and blood-borne resetting cues, at high temporal resolution and a wide concentration range. Thus, Circa-SCOPE is a powerful tool for comprehensive analysis and screening for circadian clocks’ resetting cues, and can be valuable for basic as well as translational research. Nature Publishing Group UK 2021-10-08 /pmc/articles/PMC8501123/ /pubmed/34625543 http://dx.doi.org/10.1038/s41467-021-26210-1 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/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/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Manella, Gal Aizik, Dan Aviram, Rona Golik, Marina Asher, Gad Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
title | Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
title_full | Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
title_fullStr | Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
title_full_unstemmed | Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
title_short | Circa-SCOPE: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
title_sort | circa-scope: high-throughput live single-cell imaging method for analysis of circadian clock resetting |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8501123/ https://www.ncbi.nlm.nih.gov/pubmed/34625543 http://dx.doi.org/10.1038/s41467-021-26210-1 |
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