Cold acclimation allows Drosophila flies to maintain mitochondrial functioning under cold stress

Environmental stress generally disturbs cellular homeostasis. Researchers have hypothesized that chilling injury is linked to a shortage of ATP. However, previous studies conducted on insects exposed to nonfreezing low temperatures presented conflicting results. In this study, we investigated the mitochondrial bioenergetics of Drosophila melanogaster flies exposed to chronic cold stress (4 °C). We assessed mitochondrial oxygen consumption while monitoring the rate of ATP synthesis at various times (0, 1, 2, and 3 days) during prolonged cold stress and at two assay temperatures (25 and 4 °C). We compared organelle responses between cold-susceptible and cold-acclimated phenotypes. Continuous exposure to low temperature provoked temporal declines in the rates of mitochondrial respiration and ATP synthesis. Respiratory control ratios (RCRs) suggested that mitochondria were not critically uncoupled. Nevertheless, after 3 days of continuous cold stress, a sharp decline in the mitochondrial ATP synthesis rate was observed in control flies when they were assayed at low temperature. This change was associated with reduced survival capacity in control flies. In contrast, cold-acclimated flies exhibited high survival and maintained higher rates of mitochondrial ATP synthesis and coupling (i.e., higher RCRs). Adaptive changes due to cold acclimation observed in the whole organism were thus manifested in isolated mitochondria. Our observations suggest that cold tolerance is linked to the ability to maintain bioenergetics capacity under cold stress.     

Brief carbon dioxide exposure blocks heat hardening but not cold acclimation in Drosophila melanogaster

Carbon dioxide is a commonly used anaesthetic in Drosophila research. While any detrimental effects of CO2 exposure on behaviour or traits are largely unknown, a recent study observed significant effects of CO2 exposure on rapid cold hardening and chill-coma recovery in Drosophila melanogaster. In this study we investigated the effect of a brief CO2 exposure on heat hardening and cold acclimation in D. melanogaster, measuring heat knockdown and chill-coma recovery times of flies exposed to CO2 for 1 min after hardening or acclimation. CO2 anaesthesia had a significant negative effect on heat hardening, with heat knockdown rates in hardened flies completely reduced to those of controls after CO2 exposure. Chill-coma recovery rates also significantly increased in acclimated flies that were exposed to CO2, although not to the same extent seen in the heat populations. CO2 exposure had no impact on heat knockdown rates of control flies, while there was a significant negative effect of the anaesthetic on chill-coma recovery rates of control flies. In light of these results, we suggest that CO2 should not be used after hardening in heat resistance assays due to the complete reversal of the heat hardening process upon exposure to CO2.     

Complexity of the cold acclimation response in Drosophila melanogaster

Insects can increase their resistance to cold stress when they are exposed to non-lethal conditions prior to the stress; these plastic responses are normally described only in terms of immediate effects on mortality. Here we examine in Drosophila melanogaster the short- and longer-term effects of different conditions on several measures of cold resistance, but particularly chill coma recovery. Short-term exposure to sublethal temperature (cold hardening) did not decrease chill coma recovery times even though it decreased mortality. Exposure to 12 degrees C for 2 days (acclimation) decreased chill coma recovery times for a range of stressful temperatures when flies were cultured at 25 degrees C, but did not usually affect recovery times when flies were cultured at 19 degrees C. In contrast, 2-day exposure to 12 degrees C decreased mortality regardless of rearing temperature. Rearing at 19 degrees C decreased mortality and chill coma recovery time relative to rearing at 25 degrees C. Acclimation increased the eclosion rate of eggs from stressed females, but did not affect development time or size of the offspring. These results indicate that plastic responses to cold in D. melanogaster are complex when resistance is scored in different ways, and that effects can extend across generations.     

Frostbite Protection in Mice Expressing an Antifreeze Glycoprotein

Ectotherms in northern latitudes are seasonally exposed to cold temperatures. To improve survival under cold stress, they use diverse mechanisms to increase temperature resistance and prevent tissue damage. The accumulation of anti-freeze proteins that improve cold hardiness occurs in diverse species including plants, arthropods, fish, and amphibians. We previously identified an Ixodes scapularis anti-freeze glycoprotein, named IAFGP, and demonstrated its cold protective function in the natural tick host and in a transgenic Drosophila model. Here we show, in a transgenic mouse model expressing an anti-freeze glycoprotein, that IAFGP protects mammalian cells and mice from cold shock and frostbite respectively. Transgenic skin samples showed reduced cell death upon cold storage ex vivo and transgenic mice demonstrated increased resistance to frostbite injury in vivo. IAFGP actively protects mammalian tissue from freezing, suggesting its application for the prevention of frostbite, and other diseases associated with cold exposure.     

Age-specific mortality and reproduction respond to adult dietary restriction in Drosophila melanogaster

Adult dietary yeast modulates mortality rate and reproduction of the Mediterranean fruit fly, Ceratatis capitata. In the medfly, a sugar-only diet leads to low mortality rates and reduced reproduction; addition of dietary yeast increases both mortality and egg laying. In Drosophila melanogaster low availability of dietary yeast is known to increase life span and reduce the rate of reproduction. Despite these similarities, because of differences in experimental design it remains unclear whether a common physiological mechanism modulates the effect of diet on survival. Here, we investigate how mortality rate and reproduction in D. melanogaster respond to the treatment regime used to study the medfly: no-yeast versus full diet. We find that adult medfly and D. melanogaster have opposite responses to the absence of yeast: D. melanogaster have high mortality when on no-yeast diet; when switched to full diet, D. melanogaster reduce mortality rates to the level presented by females continuously maintained on yeast. This reduction in mortality is accompanied by increased fecundity. These patterns are observed in all tested wildtype stocks, but flies made sterile by mutation in the gene oo18 RNA-binding protein (orb) lack this response. D. melanogaster, unlike medflies, appear to require adult dietary yeast to maintain maximal survival, and the capacity to assimilate yeast for somatic processes is one wildtype function of the gene orb.     

Negligible Senescence during Reproductive Dormancy in Drosophila melanogaster

Some endemic Drosophila overwinter in a state of adult reproductive diapause where egg maturation is arrested in previtellogenic stages. When maintained at cool temperatures, adult Drosophila melanogaster enter reproductive dormancy, that is, diapause or diapause-like quiescence. The ability to survive for extended periods is a typical feature of diapause syndromes. In adults this somatic persistence may involve reduced or slowed senescence. Here we assess whether reproductively dormant D. melanogaster age at slow rates. Adults were exposed to dormancy-inducing conditions for 3, 6, or 9 wk. After this period, demographic parameters were measured under normal conditions and compared to the demography of newly eclosed cohorts. The age-specific mortality rates of postdormancy adults were essentially identical to the mortality rates of newly eclosed, young flies. Postdormancy reproduction, in contrast, declined with the duration of the treatment; somatic survival during dormancy may tradeoff with later reproduction. Adults in reproductive dormancy were highly resistant to heat and to oxidative stress. Suppressed synthesis of juvenile hormone is known to regulate reproductive diapause of many insects. Treatment of dormant D. melanogaster with a juvenile hormone analog restored vitellogenesis, suppressed stress resistance, and increased demographic senescence. We conclude that D. melanogaster age at slow rates as part of their reproductive dormancy syndrome; the data do not agree with an alternative hypothesis based on heat-dependent "rate of living." We suggest that low temperature reduces neuroendocrine function, which in turn slows senescence as a function of altered stress response, nutrient reallocation, and metabolism.     

Drosophila regulate yeast density and increase yeast community similarity in a natural substrate

Drosophila melanogaster adults and larvae, but especially larvae, had profound effects on the densities and community structure of yeasts that developed in banana fruits. Pieces of fruit exposed to adult female flies previously fed fly-conditioned bananas developed higher yeast densities than pieces of the same fruits that were not exposed to flies, supporting previous suggestions that adult Drosophila vector yeasts to new substrates. However, larvae alone had dramatic effects on yeast density and species composition. When yeast densities were compared in pieces of the same fruits assigned to different treatments, fruits that developed low yeast densities in the absence of flies developed significantly higher yeast densities when exposed to larvae. Across all of the fruits, larvae regulated yeast densities within narrow limits, as compared to a much wider range of yeast densities that developed in pieces of the same fruits not exposed to flies. Larvae also affected yeast species composition, dramatically reducing species diversity across fruits, reducing variation in yeast communities from one fruit to the next (beta diversity), and encouraging the consistent development of a yeast community composed of three species of yeast (Candida californica, C. zemplinina, and Pichia kluvyeri), all of which were palatable to larvae. Larvae excreted viable cells of these three yeast species in their fecal pools, and discouraged the growth of filamentous fungi, processes which may have contributed to their effects on the yeast communities in banana fruits. These and other findings suggest that D. melanogaster adults and their larval offspring together engage in 'niche construction', facilitating a predictable microbial environment in the fruit substrates in which the larvae live and develop.     

Uncovering the benefits of fluctuating thermal regimes on cold tolerance of drosophila flies by combined metabolomic and lipidomic approach

When exposed to constant low temperatures (CLTs), insects often suffer from cumulative physiological injuries that can severely compromise their fitness and survival. Yet, mortality can be considerably lowered when the cold stress period is interrupted by periodic warm interruption(s), referred to as fluctuating thermal regimes, FTRs. In this study, we have shown that FTRs strongly promoted cold tolerance of Drosophila melanogaster adults. We then assessed whether this marked phenotypic shift was associated with detectable physiological changes, such as synthesis of cryoprotectants and/or membrane remodeling. To test these hypotheses, we conducted two different time-series Omics analyzes in adult flies submitted to CLTs vs. FTRs: metabolomics (GC/MS) and lipidomics (LC/ESI/MS) targeting membrane phospholipids. We observed increasing levels in several polyhydric alcohols (arabitol, erythritol, sorbitol, mannitol, glycerol), sugars (fructose, mannose) and amino acids (serine, alanine, glutamine) in flies under CLT. Prolonged exposure to low temperature was also associated with a marked deviation of metabolic homeostasis and warm interruptions as short as 2 h were sufficient to periodically return the metabolic system to functionality. Lipidomics revealed an increased relative proportion of phosphatidylethanolamines and a shortening of fatty acyl chains in flies exposed to cold, likely to compensate for the ordering effect of low temperature on membranes. We found a remarkable correspondence in the time-course of changes between the metabolic and phospholipids networks, both suggesting a fast homeostatic regeneration during warm intervals under FTRs. In consequence, we suggest that periodic opportunities to restore system-wide homeostasis contribute to promote cold tolerance under FTRs.     

Ethylene Induces Antifreeze Activity in Winter Rye Leaves

Antifreeze activity is induced by cold temperatures in winter rye (Secale cereale) leaves. The activity arises from six antifreeze proteins that accumulate in the apoplast of winter rye leaves during cold acclimation. The individual antifreeze proteins are similar to pathogenesis-related proteins, including glucanases, chitinases, and thaumatin-like proteins. The objective of this study was to study the regulation of antifreeze activity in response to ethylene and salicyclic acid, which are known regulators of pathogenesis-related proteins induced by pathogens. Nonacclimated plants treated with salicylic acid accumulated apoplastic proteins with no antifreeze activity. In contrast, when nonacclimated plants were exposed to ethylene, both antifreeze activity and the concentration of apoplastic protein increased in rye leaves. Immunoblotting revealed that six of the seven accumulated apoplastic proteins consisted of two glucanases, two chitinases, and two thaumatin-like proteins. The ethylene-releasing agent ethephon and the ethylene precursor 1-aminocyclopropane-1-carboxylate also induced high levels of antifreeze activity at 20 degrees C, and this effect could be blocked by the ethylene inhibitor AgNO(3). When intact rye plants were exposed to 5 degrees C, endogenous ethylene production and antifreeze activity were detected within 12 and 48 h of exposure to cold, respectively. Rye plants exposed to drought produced both ethylene and antifreeze activity within 24 h. We conclude that ethylene is involved in regulating antifreeze activity in winter rye in response to cold and drought.     

Developmental changes in Drosophila melanogaster following exposure to alternating electromagnetic fields

This study investigated the biological effects of alternating electromagnetic fields (EMFs) on developmental stages of Drosophila melanogaster eggs and the first, second and third instar larvae stages. D. melanogaster eggs and larval stages were exposed to a 11 mT 50 Hz field produced by a pair of Helmholtz coils. Each stage was exposed to aEMFs for 2, 4, 6 and 8 h. Features of adult flies such as head, thorax, abdomen and other morphological changes were studied and compared. The frequency of abnormal flies was calculated using statistical methods at P <.05. The results obtained from exposing larvae in different stages of development showed a significant increase in the number of abnormal adult flies, whereas no significant increase was observed in the group arising from eggs exposed to aEMFs. Also, it appeared that duration of exposure correlates with the increase in the number of abnormal flies. There was no significant difference in mortality rate and sex distribution of the abnormal flies between field exposed and the control groups.     

Induction of rapid cold hardening by cooling at ecologically relevant rates in Drosophila melanogaster

Over a decade ago it was hypothesized that the rapid cold hardening process allows an organism's overall cold tolerance to track changes in environmental temperature, as would occur in nature during diurnal thermal cycles. Although a number of studies have since focused on characterizing the rapid cold hardening process and on elucidating the physiological mechanisms upon which it is based, the ecological relevance of this phenomenon has received little attention. We present evidence that in Drosophila melanogaster rapid cold hardening can be induced during cooling at rates which occur naturally, and that the protection afforded in such a manner benefits the organism at ecologically relevant temperatures. Drosophila melanogaster cooled at natural rates (0.05 and 0.1 degrees C min(-1)) exhibited significantly higher survival after one hour of exposure to -7 and -8 degrees C than did those directly transferred to these temperatures or those cooled at 0.5, or 1.0 degrees C min(-1). Protection accrued throughout the cooling process (e.g., flies cooled to 0 degrees C were more cold tolerant than those cooled to 11 degrees C). Whereas D. melanogaster cooled at 1.0 degrees C min(-1) had a critical thermal minimum (i.e., the temperature at which torpor occurred) of 6.5+/-0.6 degrees C, those cooled at an ecologically relevant rate of 0.1 degrees C min(-1) had a significantly lower value of 3.9+/-0.9 degrees C.     

Consequences of heat hardening on a field fitness component in Drosophila depend on environmental temperature

Heat hardening increases thermal resistance to more extreme temperatures in the laboratory. However, heat hardening also has negative consequences, and the net benefit of hardening has not been evaluated in the field. We tested short-term heat hardening effects on the likelihood of Drosophila melanogaster to be caught at different temperatures at baits in field sites without natural resources. We predicted that hardened flies should be more frequently caught at the baits at high but not low temperatures. Under cool conditions, flies hardened at 36 degrees C, and to a lesser extent at 34 degrees C, were less frequently caught at baits than nonhardened flies a few hours after release, indicating a negative effect of hardening. In later captures, negative effects tended to disappear, particularly in males. Under warm conditions, there was an overall balance of negative and positive effects, though with a different temporal resolution. Under very hot conditions, when capture rates were low, there was a large benefit of hardening at 36 degrees C and 34 degrees C but not 33 degrees C. Finally, based on climatic records, the overall benefit of hardening in D. melanogaster is discussed as an evolved response to high temperatures occasionally experienced by organisms at some locations.     

Effects of acclimation temperature on thermal tolerance and membrane phospholipid composition in the fruit fly Drosophila melanogaster

Adaptative responses of ectothermic organisms to thermal variation typically involve the reorganization of membrane glycerophospholipids (GPLs) to maintain membrane function. We investigated how acclimation at 15, 20 and 25 degrees C during preimaginal development influences the thermal tolerance and the composition of membrane GPLs in adult Drosophila melanogaster. Long-term cold survival was significantly improved by low acclimation temperature. After 60 h at 0 degrees C, more than 80% of the 15 degrees C-acclimated flies survived while none of the 25 degrees C-acclimated flies survived. Cold shock tolerance (1h at subzero temperatures) was also slightly better in the cold acclimated flies. LT50 shifted down by ca 1.5 degrees C in 15 degrees C-acclimated flies in comparison to those acclimated at 25 degrees C. In contrast, heat tolerance was not influenced by acclimation temperature. Low temperature acclimation was associated with the increase in proportion of ethanolamine (from 52.7% to 58.5% in 25 degrees C-acclimated versus 15 degrees C-acclimated flies, respectively) at the expense of choline in GPLs. Relatively small, but statistically significant changes in lipid molecular composition were observed with decreasing acclimation temperature. In particular, the proportions of glycerophosphoethanolamines with linoleic acid (18:2) at the sn-2 position increased. No overall change in the degree of fatty acid unsaturation was observed. Thus, cold tolerance but not heat tolerance was influenced by preimaginal acclimation temperature and correlated with the changes in GPL composition in membranes of adult D. melanogaster.     

The DD2R gene expression level in the corpus allatum affects the fitness of female Drosophila melanogaster

The effect of tissue-specific suppression of the dopamine D2-like receptor gene (DD2R) in the corpus allatum (CA), the gland that synthesizes juvenile hormone (JH) on the Drosophila melanogaster resistance to heat stress has been studied. A decreased expression of the DD2R gene in the CA has been found to substantially decrease the heat stress resistance of adult transgenic female, but not male, D. melanogaster compared to the control group, this phenomenon being weakly pronounced in juvenile flies. The effect of DD2R activation on the D. melanogaster reproductive function has been estimated. It has been shown that treatment of D. melanogaster with a synthetic specific agonist of DD2R decreases the fertility, the effect being considerably stronger in adult flies than in juvenile ones. It is concluded that the change in the number of DD2Rs in CA or their activation decreases the fitness of Drosophila.     

异色瓢虫捕食胁迫对黑腹果蝇繁殖与后代发育的影响水

以模式昆虫黑腹果蝇(Drosophila melanogaster)及捕食性天敌异色瓢虫(Harmonia axyridis)为研究系统,通过把异色瓢虫接入到有黑腹果蝇的指形管中,研究了黑腹果蝇雌、雄成虫单独或共同被异色瓢虫捕食胁迫后,其自身寿命、子代的生长发育、繁殖和适合度的变化。结果表明:当雄性黑腹果蝇受到异色瓢虫成虫胁迫时,其寿命明显延长,但胁迫与否对雌性黑腹果蝇的寿命无明显影响;当黑腹果蝇的雌雄成虫同时受到胁迫时,其子代幼虫的发育历期延长,雄性后代寿命也增加,而雌性或雄性黑腹果蝇单独受到胁迫,其子代幼虫发育进度没有明显改变;当初孵黑腹果蝇幼虫受到异色瓢虫成虫直接或间接捕食胁迫后,其后续幼虫的发育历期都会受到影响,其中1龄期受到的间接捕食胁迫作用的影响大于直接捕食胁迫作用,直接捕食胁迫1龄幼虫比间接捕食胁迫更能延长其将来雌性成虫的寿命。     

大蒜素调控黑腹果蝇产卵偏嗜性和适合度

[目的]研究果蝇对大蒜素的产卵选择,并解析果蝇产卵避性的机制和生物学意义.[方法]应用产卵双向选择装置,检测黑腹果蝇Drosophila melanogaster雌成虫对0.01％,0.015％和0.02％大蒜素的产卵选择性;利用产卵装置,检测黑腹果蝇对大蒜素的位置效应;通过毛细管摄食法检测黑腹果蝇摄食行为;利用黑暗(...     

Sterols as dietary markers for Drosophila melanogaster

During cold acclimation fruit flies switch their feeding from yeast to plant food, however there are no robust molecular markers to monitor this in the wild. Drosophila melanogaster is a sterol auxotroph and relies on dietary sterols to produce lipid membranes, lipoproteins and molting hormones. We employed shotgun lipidomics to quantify eight major food sterols in total lipid extracts of heads and genital tracts of adult male and female flies. We found that their sterol composition is dynamic and reflective of fly diet in an organ-specific manner. Season-dependent changes observed in the organs of wild-living flies suggested that the molar ratio between yeast (ergosterol, zymosterol) and plant (sitosterol, stigmasterol) sterols is a quantifiable, generic and unequivocal marker of their feeding behavior suitable for ecological and environmental population-based studies. The enrichment of phytosterols over yeast sterols in wild-living flies at low temperatures is consistent with switching from yeast to plant diet and corroborates the concomitantly increased unsaturation of their membrane lipids.     

Characterization of antifreeze activity in Antarctic plants

Deschampsia antarctica and Colobanthus quitensis are the only vascular plants to have colonized the Maritime Antarctic, which is characterized by its permanently low temperature and frequent summer frosts. To understand how the plants survive freezing temperatures year-round, antifreeze activity was assayed in apoplastic extracts obtained from both non-acclimated and cold-acclimated Antarctic plants. By observing the shape of ice crystals grown in dilution series of the extracts, it was found that D. antarctica had antifreeze activity, but C. quitensis did not. D. antarctica exhibited antifreeze activity in the non-acclimated state and this activity increased after cold acclimation. The antifreeze activity in D. antarctica was labile to proteolysis and high temperature, active over a wide pH range, and associated with molecules greater than 10 kDa in molecular weight. These results show that D. antarctica produces antifreeze proteins that are secreted into the apoplast. When examined by SDS-PAGE, the apoplastic extracts from cold-acclimated D. antarctica exhibited 13 polypeptides. It is concluded that D. antarctica accumulates AFPs as part of its mechanism of freezing tolerance. Moreover, this is the first plant in which antifreeze activity has been observed to be constitutive.     

Investigating thermal acclimation effects before and after a cold shock in Drosophila melanogaster using behavioural assays

Acclimation to environmental change can impose both costs and benefits to organisms. In this study we explored to what extent locomotor behaviour of Drosophila melanogaster is influenced by developmental temperature and adult temperature in both the laboratory and the field. Following development at 15, 25, or 31 °C, adult flies were tested for locomotor activity at all developmental temperatures in the laboratory before and after exposure to a cold shock and in the field for their ability to locate resources after a cold shock. Both test (15, 25, and 31 °C) and developmental temperatures strongly affected locomoter activity, with flies developed at 25 °C having the highest activity at all three test temperatures before the cold shock. After the cold shock flies developed at 15 °C had higher activity compared with flies developed at 25 and 31 °C when tested at 15 and 25 °C, and flies developed at 25 °C had the highest activity when tested at 31 °C. Furthermore, flies developed at 31 °C showed longer recovery times following the cold shock at test temperatures of 15 and 25 °C. However, flies acclimated at 15 °C during development did not recover faster at 15 and 25 °C compared with flies developed at 25 °C. There were no significant correlations between recovery time and locomotor activity at any of the test temperatures. Flies developed at 15 °C and exposed to a cold shock before release in the field were much more successful in locating a resource at low field temperatures compared with flies developed at 25 and 31 °C. Our results provide support for both the beneficial acclimation hypothesis and the optimal developmental temperature hypothesis, but the results are highly context dependent and change with the temperature experienced by the individual during its lifetime.