Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells
Abstract Pv11 is an insect cell line established from the midge Polypedilum vanderplanki, whose larval form exhibits an extreme desiccation tolerance known as anhydrobiosis. Pv11 itself is also capable of anhydrobiosis, which is induced by trehalose treatment. Here we report the successful construct...
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doaj-e01e98b6a5b0485fa92412c59cfd95332021-10-10T11:28:02ZengNature Publishing GroupScientific Reports2045-23222021-10-0111111410.1038/s41598-021-98905-wCas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cellsYugo Miyata0Hiroto Fuse1Shoko Tokumoto2Yusuke Hiki3Ruslan Deviatiiarov4Yuki Yoshida5Takahiro G. Yamada6Richard Cornette7Oleg Gusev8Elena Shagimardanova9Akira Funahashi10Takahiro Kikawada11Division of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO)Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of TokyoDepartment of Integrated Biosciences, Graduate School of Frontier Sciences, The University of TokyoDepartment of Biosciences and Informatics, Keio UniversityExtreme Biology Laboratory, Institute of Fundamental Medicine and Biology, Kazan Federal UniversityInstitute for Advanced Biosciences, Keio UniversityDepartment of Biosciences and Informatics, Keio UniversityDivision of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO)Extreme Biology Laboratory, Institute of Fundamental Medicine and Biology, Kazan Federal UniversityExtreme Biology Laboratory, Institute of Fundamental Medicine and Biology, Kazan Federal UniversityDepartment of Biosciences and Informatics, Keio UniversityDivision of Biomaterial Sciences, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization (NARO)Abstract Pv11 is an insect cell line established from the midge Polypedilum vanderplanki, whose larval form exhibits an extreme desiccation tolerance known as anhydrobiosis. Pv11 itself is also capable of anhydrobiosis, which is induced by trehalose treatment. Here we report the successful construction of a genome editing system for Pv11 cells and its application to the identification of signaling pathways involved in anhydrobiosis. Using the Cas9-mediated gene knock-in system, we established Pv11 cells that stably expressed GCaMP3 to monitor intracellular Ca2+ mobilization. Intriguingly, trehalose treatment evoked a transient increase in cytosolic Ca2+ concentration, and further experiments revealed that the calmodulin–calcineurin–NFAT pathway contributes to tolerance of trehalose treatment as well as desiccation tolerance, while the calmodulin–calmodulin kinase–CREB pathway conferred only desiccation tolerance on Pv11 cells. Thus, our results show a critical contribution of the trehalose-induced Ca2+ surge to anhydrobiosis and demonstrate temporally different roles for each signaling pathway.https://doi.org/10.1038/s41598-021-98905-w |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yugo Miyata Hiroto Fuse Shoko Tokumoto Yusuke Hiki Ruslan Deviatiiarov Yuki Yoshida Takahiro G. Yamada Richard Cornette Oleg Gusev Elena Shagimardanova Akira Funahashi Takahiro Kikawada |
spellingShingle |
Yugo Miyata Hiroto Fuse Shoko Tokumoto Yusuke Hiki Ruslan Deviatiiarov Yuki Yoshida Takahiro G. Yamada Richard Cornette Oleg Gusev Elena Shagimardanova Akira Funahashi Takahiro Kikawada Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells Scientific Reports |
author_facet |
Yugo Miyata Hiroto Fuse Shoko Tokumoto Yusuke Hiki Ruslan Deviatiiarov Yuki Yoshida Takahiro G. Yamada Richard Cornette Oleg Gusev Elena Shagimardanova Akira Funahashi Takahiro Kikawada |
author_sort |
Yugo Miyata |
title |
Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells |
title_short |
Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells |
title_full |
Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells |
title_fullStr |
Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells |
title_full_unstemmed |
Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells |
title_sort |
cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in pv11 cells |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2021-10-01 |
description |
Abstract Pv11 is an insect cell line established from the midge Polypedilum vanderplanki, whose larval form exhibits an extreme desiccation tolerance known as anhydrobiosis. Pv11 itself is also capable of anhydrobiosis, which is induced by trehalose treatment. Here we report the successful construction of a genome editing system for Pv11 cells and its application to the identification of signaling pathways involved in anhydrobiosis. Using the Cas9-mediated gene knock-in system, we established Pv11 cells that stably expressed GCaMP3 to monitor intracellular Ca2+ mobilization. Intriguingly, trehalose treatment evoked a transient increase in cytosolic Ca2+ concentration, and further experiments revealed that the calmodulin–calcineurin–NFAT pathway contributes to tolerance of trehalose treatment as well as desiccation tolerance, while the calmodulin–calmodulin kinase–CREB pathway conferred only desiccation tolerance on Pv11 cells. Thus, our results show a critical contribution of the trehalose-induced Ca2+ surge to anhydrobiosis and demonstrate temporally different roles for each signaling pathway. |
url |
https://doi.org/10.1038/s41598-021-98905-w |
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