TY - JOUR
T1 - Cas9-mediated genome editing reveals a significant contribution of calcium signaling pathways to anhydrobiosis in Pv11 cells
AU - Miyata, Yugo
AU - Fuse, Hiroto
AU - Tokumoto, Shoko
AU - Hiki, Yusuke
AU - Deviatiiarov, Ruslan
AU - Yoshida, Yuki
AU - Yamada, Takahiro G.
AU - Cornette, Richard
AU - Gusev, Oleg
AU - Shagimardanova, Elena
AU - Funahashi, Akira
AU - Kikawada, Takahiro
N1 - Funding Information:
We would like to express our deep gratitude to Tomoe Shiratori (NARO) for her technical support. This work was supported by Grants-in-Aid for Scientific Research (KAKENHI) Grants (numbers JP25128714 and JP17H01511 to T.K., JP18K14472 to Y.M., JP19J12030 to S.T., JP18J21155 to Y.Y., JP16K07308 to R.C. and JP18H02217 to O.G.), and T.K. and R.C. were also funded by the Agriculture, Forestry and Fisheries Research Council of the Ministry of Agriculture, Forestry and Fisheries (http://www.affrc.maff.go.jp) grant “Strategic International Collaborative Research Project” (JPJ008837). E.S. and R.D. were supported by Russian Science Foundation grant 20-44-07002 for the bioinformatics part of the study.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - 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.
AB - 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.
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U2 - 10.1038/s41598-021-98905-w
DO - 10.1038/s41598-021-98905-w
M3 - Article
C2 - 34611198
AN - SCOPUS:85116372183
VL - 11
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
IS - 1
M1 - 19698
ER -