Lipophilic contaminants influence cold tolerance of invertebrates through changes in cell membrane fluidity.

Contaminants taken up by living organisms in the environment as a result of anthropogenic contamination can reduce the tolerance of natural stressors, e.g., low temperatures, but the physiological mechanisms behind these interactions of effects are poorly understood. The tolerance to low temperatures of organisms that cannot regulate their body temperature (ectotherms) depends on their ability to increase the fluidity of their cellular membranes at low temperatures. Our study shows that contaminants accumulating in lipids of organisms alter the physical state of their membranes simply by being present. Contaminants of varying chemical structures can alter the membrane fluidity in either direction and correspondingly modulate the cold tolerance of intact animals.

Linking membrane physical properties and low temperature tolerance in arthropods.

Maintenance of membrane fluidity is of crucial importance in ectotherms experiencing thermal changes. This maintenance has in ectotherms most often been indicated using indirect measures of biochemical changes of phospholipid membranes, which is then assumed to modulate the physico-chemical properties of the membrane. Here, we measure bending rigidity characterizing the membrane flexibility of re-constituted membrane vesicles to provide a more direct link between membrane physical characteristics and low temperature tolerance. Bending rigidity of lipid bilayers was measured in vitro using Giant Unilamellar Vesicles formed from phospholipid extracts of the springtail, Folsomia candida. The bending rigidity of these membranes decreased when exposed to 0.4 vol% ethanol (0.23 mM/L). Springtails exposed to ethanol for 24h significantly increased their cold shock tolerance. Thus, by chemically inducing decreased membrane rigidity, we have shown a direct link between the physico-chemical properties of the membranes and the capacity to tolerate low temperature in a chill-susceptible arthropod.

Induced cold-tolerance mechanisms depend on duration of acclimation in the chill-sensitive Folsomia candida (Collembola).

During cold periods ectotherms may improve low temperature tolerance via rapid cold hardening (RCH) over a period of hours and/or long-term cold acclimation (LTCA) during days, weeks or months. However, the effect of duration and the major underlying mechanisms of these processes are still not fully understood. In the present study, the molecular and biochemical responses to RCH (1-3 h) and LTCA (1-3 days) and the corresponding benefits to survival were investigated using the chill-sensitive collembolan Folsomia candida. We investigated osmolyte accumulation, membrane restructuring and transcription of candidate genes as well as survival benefits in response to RCH and LTCA. RCH induced significant upregulation of targeted genes encoding enzymes related to carbohydrate metabolic pathways and genes encoding small and constitutively expressed heat shock proteins (Hsps), indicating that the animals rely on protein protection from a subset of Hsps during RCH and probably also LTCA. The upregulation of genes involved in carbohydrate metabolic processes initiated during RCH was likely responsible for a transient accumulation of myoinositol during LTCA, which may support the protection of protein and membrane function and structure. Membrane restructuring, composed especially of a significantly increased ratio of unsaturated to saturated phospholipid fatty acids seems to be a mechanism supplementary to activation of Hsps and myoinositol accumulation in LTCA. Thus, the moderate increase in cold shock tolerance conferred by RCH seems to be dominated by effects of Hsps, whereas the substantially better cold tolerance achieved after LTCA is dominated by post-transcriptional processes increasing membrane fluidity and cryoprotectant concentration.

Ecological and molecular consequences of prolonged drought and subsequent rehydration in Folsomia candida (Collembola).

Drought tolerance in water-permeable, soil-living Collembola (e.g. Folsomia candida) is achieved due to a unique water vapour absorption mechanism, where accumulation of sugars and polyols is essential. However, the molecular mechanisms underlying such adaptation as well as the maintenance of this survival strategy and the responses to rehydration after prolonged drought in these soil-living Collembola are unclear. In the present study, the functional relationships between ecological drought responses and expression of related target genes were investigated in F. candida exposed to mild and severe drought for up to 5 weeks by relating survival, moulting and reproduction rate with mRNA-level expression of 7 target genes during drought, dehydration and rehydration. Prolonged drought and subsequent rehydration induced significant changes in gene expression which could be related to the fitness traits studied. In F. candida the ecological and molecular responses to mild drought differed from those of severe drought. From the changes in gene expression, where significantly increased expression of Glucose-6-phosphate-isomerase (gpi) and Heat shock protein 70 (hsp70) was dominating, it is proposed that protection of cellular structure and function during prolonged mild drought (98.2% RH) is partly achieved from a continuous accumulation of compatible osmolytes in F. candida. To achieve protection during and after prolonged severe drought (96.1% RH), components related to cell division and development such as inositol monophosphatase and one of the small heat shock proteins (sHsps), Heat shock protein23 (hsp23), seem to play an important role in F. candida.

Body metal concentrations and glycogen reserves in earthworms (Dendrobaena octaedra) from contaminated and uncontaminated forest soil.

Stress originating from toxicants such as heavy metals can induce compensatory changes in the energy metabolism of organisms due to increased energy expenses associated with detoxification and excretion processes. These energy expenses may be reflected in the available energy reserves such as glycogen. In a field study the earthworm, Dendrobaena octaedra, was collected from polluted areas, and from unpolluted reference areas. If present in the environment, cadmium, lead and copper accumulated to high concentrations in D. octaedra. In contrast, other toxic metals such as aluminium, nickel and zinc appeared to be regulated and kept at low internal concentrations compared to soil concentrations. Lead, cadmium and copper accumulation did not correlate with glycogen reserves of individual worms. In contrast, aluminium, nickel and zinc were negatively correlated with glycogen reserves. These results suggest that coping with different metals in earthworms is associated with differential energy demands depending on the associated detoxification strategy.

Hsp70 expression and metabolite composition in response to short-term thermal changes in Folsomia candida (Collembola).

In the present study the joint transcriptomic and metabolomic responses in Folsomia candida (Collembola) to temperature changes on a short-term scale were studied. Change in heat tolerance was examined as survival after a 35 degrees C heat shock (2h) in the course of either a fluctuating temperature regime (8 to 32 degrees C; pre-treated) or a constant temperature (20 degrees C; control) over a period of 24h. Exposure to a temperature increase from 20 to 32 degrees C (2.4 degrees C min(-)(1)) induced a significantly increased heat tolerance which continued throughout the experiment. Expression of the gene encoding heat shock protein Hsp70 was assessed at the mRNA level using real time quantitative polymerase chain reaction (QPCR). Hsp70 was rapidly induced and significantly increased by the temperature increase. The relative concentrations of low molecular weight metabolites were analysed in F. candida using nuclear magnetic resonance spectroscopy ((1)H NMR). A significant metabolomic divergence between pre-treated and control collembolans was evident; partly due to a significantly reduced relative concentration of five free amino acids (arginine, leucine, lysine, phenylalanine and tyrosine) in pre-treated collembolans. These results, obtained from ecological transcriptomics and metabolomics jointly generated insight on various levels into the combined responses to a changing environment.