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A mixture of innate cryoprotectants is key for freeze tolerance and cryopreservation of a drosophilid fly larva
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SYSNO ASEP 0557907 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title A mixture of innate cryoprotectants is key for freeze tolerance and cryopreservation of a drosophilid fly larva Author(s) Kučera, Lukáš (UMG-J)
Moos, Martin (BC-A) RID, ORCID
Štětina, Tomáš (BC-A) ORCID
Korbelová, Jaroslava (BC-A)
Vodrážka, Petr (BC-A) ORCID
Des Marteaux, Lauren E. (BC-A) RID
Grgac, Robert (BC-A)
Hůla, Petr (BC-A)
Rozsypal, Jan (BC-A) RID, ORCID
Faltus, M. (CZ)
Šimek, Petr (BC-A) RID, ORCID
Sedláček, Radislav (UMG-J) RID
Košťál, Vladimír (BC-A) RID, ORCIDNumber of authors 13 Article number jeb243934 Source Title Journal of Experimental Biology. - : Company of Biologists - ISSN 0022-0949
Roč. 225, č. 8 (2022)Number of pages 25 s. Publication form Online - E Language eng - English Country GB - United Kingdom Keywords Freeze tolerance ; Cryoprotection ; Insects ; Metabolites ; Natural deep eutectic systems Subject RIV EB - Genetics ; Molecular Biology OECD category Cell biology Subject RIV - cooperation Biology Centre (since 2006) - Physiology R&D Projects GA19-13381S GA ČR - Czech Science Foundation (CSF) GA17-22276S GA ČR - Czech Science Foundation (CSF) LM2018126 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) EF16_013/0001789 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Limited access Institutional support UMG-J - RVO:68378050 ; BC-A - RVO:60077344 UT WOS 000794016900015 DOI 10.1242/jeb.243934 Annotation Insects that naturally tolerate internal freezing produce complex mixtures of multiple cryoprotectants (CPs). Better knowledge on composition of these mixtures, and on the mechanisms of individual CP interactions, could inspire development of laboratory CP formulations optimized for cryopreservation of cells and other biological material. Here, we identify and quantify (using high resolution mass spectrometry) a range of putative CPs in larval tissues of a subarctic fly, Chymomyza costata, which survives long-term cryopreservation in liquid nitrogen. The CPs proline, trehalose, glutamine, asparagine, glycine betaine, glycerophosphoethanolamine, glycerophosphocholine and sarcosine accumulate in hemolymph in a ratio of 313:108:55:26:6:4:2.9:0.5 mmol l(-1). Using calorimetry, we show that artificial mixtures, mimicking the concentrations of major CPs in hemolymph of freeze-tolerant larvae, suppress the melting point of water and significantly reduce the ice fraction. We demonstrate in a bioassay that mixtures of CPs administered through the diet act synergistically rather than additively to enable cryopreservation of otherwise freeze-sensitive larvae. Using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), we show that during slow extracellular freezing trehalose becomes concentrated in partially dehydrated hemolymph where it stimulates transition to the amorphous glass phase. In contrast, proline moves to the boundary between extracellular ice and dehydrated hemolymph and tissues where it probably forms a layer of dense viscoelastic liquid. We propose that amorphous glass and viscoelastic liquids may protect macromolecules and cells from thermomechanical shocks associated with freezing and transfer into and out of liquid nitrogen. Workplace Institute of Molecular Genetics Contact Nikol Škňouřilová, nikol.sknourilova@img.cas.cz, Tel.: 241 063 217 Year of Publishing 2023 Electronic address https://journals.biologists.com/jeb/article-abstract/225/8/jeb243934/275162/A-mixture-of-innate-cryoprotectants-is-key-for?redirectedFrom=fulltext
Number of the records: 1