Cells from an anhydrobiotic chironomid survive almost complete desiccation

Nakahara, Yuko; Imanishi, Shigeo; Mitsumasu, Kanako; Kanamori, Yasushi; Iwata, Ken-ichi; Watanabe, Masahiko; Kikawada, Takahiro; Okuda, Takashi

doi:10.1016/j.cryobiol.2009.10.004

Cited by 40 publications

(42 citation statements)

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“…The latter category represents fat body cells and the former might be non-differentiated cells of hematopoietic organs or imaginal pads. These cells in fat body of the larvae have been shown earlier to be capable of surviving complete desiccation even if dissected from the larvae, which makes this cell type a convenient model for studies of anhydrobiosis [37], [38].…”

Section: Resultsmentioning

confidence: 99%

Anhydrobiosis-Associated Nuclear DNA Damage and Repair in the Sleeping Chironomid: Linkage with Radioresistance

et al. 2010

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Anhydrobiotic chironomid larvae can withstand prolonged complete desiccation as well as other external stresses including ionizing radiation. To understand the cross-tolerance mechanism, we have analyzed the structural changes in the nuclear DNA using transmission electron microscopy and DNA comet assays in relation to anhydrobiosis and radiation. We found that dehydration causes alterations in chromatin structure and a severe fragmentation of nuclear DNA in the cells of the larvae despite successful anhydrobiosis. Furthermore, while the larvae had restored physiological activity within an hour following rehydration, nuclear DNA restoration typically took 72 to 96 h. The DNA fragmentation level and the recovery of DNA integrity in the rehydrated larvae after anhydrobiosis were similar to those of hydrated larvae irradiated with 70 Gy of high-linear energy transfer (LET) ions (4He). In contrast, low-LET radiation (gamma-rays) of the same dose caused less initial damage to the larvae, and DNA was completely repaired within within 24 h. The expression of genes encoding the DNA repair enzymes occurred upon entering anhydrobiosis and exposure to high- and low-LET radiations, indicative of DNA damage that includes double-strand breaks and their subsequent repair. The expression of antioxidant enzymes-coding genes was also elevated in the anhydrobiotic and the gamma-ray-irradiated larvae that probably functions to reduce the negative effect of reactive oxygen species upon exposure to these stresses. Indeed the mature antioxidant proteins accumulated in the dry larvae and the total activity of antioxidants increased by a 3–4 fold in association with anhydrobiosis. We conclude that one of the factors explaining the relationship between radioresistance and the ability to undergo anhydrobiosis in the sleeping chironomid could be an adaptation to desiccation-inflicted nuclear DNA damage. There were also similarities in the molecular response of the larvae to damage caused by desiccation and ionizing radiation.

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Section: Resultsmentioning

confidence: 99%

Anhydrobiosis-Associated Nuclear DNA Damage and Repair in the Sleeping Chironomid: Linkage with Radioresistance

et al. 2010

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“…To gain further understanding of PvLEA protein functions in P. vanderplanki , we performed an RNA-seq analysis of the Pv11 cell line established from embryonic stem cells of P. vanderplanki and capable of entering anhydrobiosis (5,6). We also investigated patterns of intracellular localization in the Sf9 cell line.…”

Section: Introductionmentioning

confidence: 99%

Intracellular localization and gene expression analysis provide new insights on LEA proteins’ diversity in anhydrobiotic midge

Несмелов

et al. 2019

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20Anhydrobiosis, an adaptive ability to withstand complete desiccation, is in insects 21 limited to a single species: the nonbiting midge Polypedilum vanderplanki (the sleeping 22 chironomid). Evolution of anhydrobiosis in a single representative of a large genus is 23 associated with drastic changes in genome structure, including the emergence of new 24 multimember gene families directly involved in desiccation tolerance. Among them, Late 25 Embryogenesis Abundant (LEA) proteins, which protect other proteins from aggregation 26 caused by desiccation, are believed to originate via horizontal gene transfer from a bacterial 27 donor. To obtain new insights on the biological background of the expanded 27-member LEA 28 protein group in P. vanderplanki, we investigated the expression of corresponding genes in a 29 P. vanderplanki-derived cell line, capable of anhydrobiosis, in a normal state and during 30 induction of desiccation tolerance. We found that all LEA proteins genes identified in P.31 vanderplanki's genome except PvLea16 and PvLea17 are also expressed in Pv11 cells. Their 32 expression was elevated in response to anhydrobiosis-inducing trehalose treatment. 33 Expression patterns of PvLea genes were well preserved in Pv11 cells in comparison to P. 34 vanderplanki's larvae both in the control group and during the anhydrobiosis cycle. We also 35 investigated localization of LEA proteins in Pv11 cells and Sf9 cells and found a different 36 level of conservation in intracellular localization of the protein expressed in mammalian and 37 insect cells. 38 39 Introduction 40 One of the most intriguing adaptations to extreme environments is the phenomenon of 41 anhydrobiosis-the ability to withstand complete desiccation in a nonmetabolic state (see 42 review in (1)-which allows living organisms to revive after complete desiccation. 3 43 Organisms capable of entering anhydrobiosis are found across different taxa, and the larvae 44 of the chironomid Polypedilum vanderplanki (Insecta, Diptera) are one of the most complex 45 of these from an evolutionary point of view, as well as being a unique case of emergence of 46 anhydrobiosis in a single species of eukaryotes (2). 47 For the larvae, entering the anhydrobiotic state takes 48 hours and is mediated by 48 several key events including replacement of water with trehalose and vitrification, as well as 49 accumulation of protective biomolecules, including heat shock proteins, antioxidants and 50 enzymes, aquaporins, and Late Embryogenesis Abundant (LEA) proteins (2,3). The latter are 51 highly hydrophilic proteins, well known for their protective effect against water deficits, and 52 they are found in anhydrobiotic organisms in both the plant and animal kingdoms (2). 53 Previously, we identified 27 genes of LEA proteins (PvLea) in the P. vanderplanki 54 genome, all of which are encoding members of LEA group 3. Many of these genes become 55 induced during desiccation in larvae (4). LEA proteins are believed to be one of the main 56 participants in anhydrobiosis mechanisms, ...

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“…In 2010, the Pv11 cell line was established as an embryonic cell culture from P. vanderplanki 9 . Desiccation tolerance of Pv11 cells is induced by treatment with culture medium containing 600 mM trehalose for 48 h. Even after dehydration in a desiccator (<10% relative humidity) for 12 days and rehydration for 1 h, the trehalose-treated Pv11 cells were able to resume proliferation 10 , whereas the other insect cell lines (Sf9, BmN-4, AeAl-2, AnCu-35, and S2) did not proliferate after rehydration.…”

Section: Introductionmentioning

confidence: 99%

Transcriptome analysis of Pv11 cells infers the mechanism of desiccation tolerance and recovery

Yamada¹,

Suetsugu

Deviatiiarov

et al. 2018

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The larvae of the African midge, Polypedilum vanderplanki, can enter an ametabolic state called anhydrobiosis to conquer fatal desiccation stress. The Pv11 cell line, derived from embryos of the midge, shows desiccation tolerance by pretreatment with trehalose before desiccation; they can resume proliferation after rehydration. To address the underlying molecular mechanisms, we desiccated Pv11 cells after pretreatment with the medium containing trehalose and induced proliferation by rehydration. We collected the cells at each before and after desiccation and rehydration step and performed CAGE-seq of mRNA of those cells. By analysing differentially expressed genes (DEGs) among the results of CAGE-seq, we detected 384 DEGs after trehalose treatment and 14 DEGs after rehydration. Hierarchical clustering of the identified DEGs indicated that rehydration returns their expression pattern to that in the control culture state. DEGs involved in various stress responses, detoxification of harmful chemicals, and regulation of oxidoreduction were upregulated by trehalose treatment. DEGs for rehydration supported that DNA repair is one of the potential mechanisms involves recovery. This study provided initial insight into the molecular mechanisms underlying the extreme desiccation tolerance of Pv11 cells with a potential for proliferation following rehydration.trehalose-treated, desiccated, and rehydrated Pv11 cells, followed by gene ontology (GO) analysis of the identified DEGs. To the best of our knowledge, this is the first report of comprehensive DEG analysis using CAGE-seq data for the Pv11 cells that infers a putative mechanism of their avoidance of cell death and recovery from anhydrobiosis.

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Cells from an anhydrobiotic chironomid survive almost complete desiccation

Cited by 40 publications

References 40 publications

Anhydrobiosis-Associated Nuclear DNA Damage and Repair in the Sleeping Chironomid: Linkage with Radioresistance

Anhydrobiosis-Associated Nuclear DNA Damage and Repair in the Sleeping Chironomid: Linkage with Radioresistance

Intracellular localization and gene expression analysis provide new insights on LEA proteins’ diversity in anhydrobiotic midge

Transcriptome analysis of Pv11 cells infers the mechanism of desiccation tolerance and recovery

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