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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Autores principales:	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
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e01e98b6a5b0485fa92412c59cfd9533
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spelling	oai:doaj.org-article:e01e98b6a5b0485fa92412c59cfd95332021-12-02T19:16:19ZCas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells10.1038/s41598-021-98905-w2045-2322https://doaj.org/article/e01e98b6a5b0485fa92412c59cfd95332021-10-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-98905-whttps://doaj.org/toc/2045-2322Abstract 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.Yugo MiyataHiroto FuseShoko TokumotoYusuke HikiRuslan DeviatiiarovYuki YoshidaTakahiro G. YamadaRichard CornetteOleg GusevElena ShagimardanovaAkira FunahashiTakahiro KikawadaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-14 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q 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
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.
format	article
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
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 Portfolio
publishDate	2021
url	https://doaj.org/article/e01e98b6a5b0485fa92412c59cfd9533
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Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells

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