Abstract
The cryopreservation techniques proposed for embryos of the fruit fly Drosophila melanogaster are not yet ready for practical use. Alternative methods for long-term storage of D. melanogaster strains, although urgently needed, do not exist. Herein, we describe a narrow interval of low temperatures under which the larvae of D. melanogaster can be stored in quiescence for up to two months. The development of larvae was arrested at the pre-wandering stage under fluctuating thermal regime (FTR), which simultaneously resulted in diminishing the accumulation of indirect chill injuries. Our physiological, metabolomic, and transcriptomic analyses revealed that compared to larvae stored at constant low temperatures, the larvae stored under FTR conditions were able to decrease the rates of depletion of energy substrates, exploited brief warm episodes of FTR for homeostatic control of metabolite levels, and more efficiently exerted protection against oxidative damage.
Highlights
The cryopreservation techniques proposed for embryos of the fruit fly Drosophila melanogaster are not yet ready for practical use
Based on previous reports[18,19] and our previous results[11], we focused on constant low temperatures (CLTs) in the range from +3 to +9 °C and assessed the time limits of larval storability
The ‘temperature-window’ for successful quiescence is theoretically delimited by the temperature of the upper limit of cold injury zone (ULCIZ) from below[21] and the temperature of the lower developmental threshold (LDT) from above[22]
Summary
The cryopreservation techniques proposed for embryos of the fruit fly Drosophila melanogaster are not yet ready for practical use. A strategy of long-term storage at temperatures slightly below the threshold for development, exploiting the natural features of insect diapause, presents itself as a useful alternative to cryopreservation in many insects[1]. In insect species that lack diapause, the potential for practical long-term storage at low temperatures could be limited unless the host of problems linked to development of indirect chill injury is solved. Using a combination of low temperature-induced quiescence and FTR-based suppression of chill injuries, we were able to store viable larvae for up to 2 months. Depletion of energy substrates, loss of metabolite homeostasis and oxidative damage as potential mechanisms responsible for accumulation of indirect chill injury, which sets physiological limits on D. melanogaster low-temperature storability
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