Dessication Stress

The threat of desiccation for organisms inhabiting the intertidal zone occurs during emersion at low tides or when organisms are positioned in the high intertidal zone, where wetting occurs primarily by spring tides, storm waves, and spray. Drying due to evaporative water loss is the most common mechanism for dehydration, although during winter in northern temperate regions freezing can also occur, which reduces the liquid water in extracellular fluids and can lead to intracellular dehydration in multicellular organisms. Freezing tolerance has been reported and characterized for a number of intertidal invertebrates, including gastropods such as an air-breathing snail and a periwinkle, and bivalve genera including the common and ribbed mussels

Role of Intrinsic Disorder in Animal Desiccation Tolerance

This review compares the molecular strategies employed by anhydrobiotic invertebrates to survive extreme water stress. Intrinsically disordered proteins (IDPs) play a central role in desiccation tolerance in all species investigated. Various hypotheses about the functions of anhydrobiosisrelated intrinsically disordered (ARID) proteins, including late embryogenesis abundant (LEA) and tardigrade-specific intrinsically disordered proteins, were evaluated by broad sequence characterization. A surprisingly wide range in sequence characteristics including hydropathy and the frequency and distribution of charges was discovered. Interestingly, two clusters of similar proteins were found that potentially correlate with distinct functions. This may indicate two broad groups of ARID proteins, composed of one group that folds into functional conformations during desiccation and a second group that potentially displays functions in the hydrated state. A broad range of physiochemical properties suggest that folding may be induced by factors such as hydration level, molecular crowding, and interactions with binding partners. This plasticity may be required to fine tune the ARID-proteome response at different hydration levels during desiccation. Furthermore, the sequence properties of some LEA proteins share qualities with IDPs known to undergo liquid-liquid phase separations during environmental challenges

Concert recording 2015-11-23

[Track 01]. Sonata in C major BWV 1033. Andante ; [Track 02]. Allegro ; [Track 03]. Adagio ; [Track 04]. Menuet I ; Menuet II / J.S. Bach -- [Track 05]. Mawal for solo flute / Avi Eilam-Amzallag -- [Track 06]. 2 flutes in 3 scenes. Burghers ; [Track 07]. There were daffodils ; [Track 08]. By royal coach / Gary Schocker -- [Track 09]. Andante and allegretto from Sonata for flute and piano / General John Reid -- [Track 10]. Fantasie / Gabriel Fauré

Reduced mitochondrial efficiency explains mismatched growth and metabolic rate at supraoptimal temperatures.

The relationship between whole-organism growth and metabolism is generally assumed to be positive and causative; higher metabolic rates support higher growth rates. In Manduca sexta, existing data demonstrate a deviation from this simple prediction: at supraoptimal temperatures for larval growth, metabolic rate keeps increasing while growth rate is decreasing. This mismatch presumably reflects the rising “cost of maintenance” with temperature. Precisely what constitutes this cost is not clear, but we suspect the efficiency with which mitochondria harness oxygen and organic substrates into cellular energy (ATP) is key. We tested this by integrating existing data on M. sexta growth and metabolism with new data on mitochondrial bioenergetics across the temperature range 14°–42°C. Across this range, our measure of mitochondrial efficiency closely paralleled larval growth rates. At supraoptimal temperatures for growth, mitochondrial efficiency was reduced, which could explain the mismatch between growth and metabolism observed at the whole-organism level

Mitochondrial energetics of benthic and pelagic Antarctic teleosts.

Antarctic fauna are highly adapted to the frigid waters of the Southern Ocean. This study describes the in vitro temperature sensitivity of oxygen consumption rates measured in liver mitochondria from the pelagic notothenioid Pleuragramma antarcticum between 5 and 35 C. Oxygen fluxes were measured after the addition of millimolar levels of pyruvate, malate, succinate and glutamate (state II, LEAK) and saturating levels of ADP [state III, oxidative phosphorylation (OXPHOS)]. State III respiration significantly decreased above 18.7 C. A comparison of the oxidative capacities among P. antarcticum and other notothenioids showed significant differences in state III respiration, where benthic species exhibited about 50 % lower rates than P. antarcticum . In addition, state III respiration rates normalized per milligram of mitochondrial protein of P. antarcticum were up to eight times higher than state III rates reported in the literature for other notothenioids. The comparatively high respiration rates measured in this study may be explained by our approach, which engaged both complexes I and II under conditions of oxidative phosphorylation. State III rates of independently activated complexes I and II were found to range from 42 to 100 % of rates obtained when both complexes were activated simultaneously in the same species. The remarkable tolerance of P. antarcticum OXPHOS toward warmer temperatures was unexpected for an Antarctic stenotherm and may indicate that thermal sensitivity of their mitochondria is not the driving force behind their stenothermy

Mitochondrial Energetics of Benthic and Pelagic Antarctic Teleosts

Antarctic fauna are highly adapted to the frigid waters of the Southern Ocean. This study describes the in vitro temperature sensitivity of oxygen consumption rates measured in liver mitochondria from the pelagic notothenioid Pleuragramma antarcticum between 5 and 35 C. Oxygen fluxes were measured after the addition of millimolar levels of pyruvate, malate, succinate and glutamate (state II, LEAK) and saturating levels of ADP [state III, oxidative phosphorylation (OXPHOS)]. State III respiration significantly decreased above 18.7 C. A comparison of the oxidative capacities among P. antarcticum and other notothenioids showed significant differences in state III respiration, where benthic species exhibited about 50 % lower rates than P. antarcticum . In addition, state III respiration rates normalized per milligram of mitochondrial protein of P. antarcticum were up to eight times higher than state III rates reported in the literature for other notothenioids. The comparatively high respiration rates measured in this study may be explained by our approach, which engaged both complexes I and II under conditions of oxidative phosphorylation. State III rates of independently activated complexes I and II were found to range from 42 to 100 % of rates obtained when both complexes were activated simultaneously in the same species. The remarkable tolerance of P. antarcticum OXPHOS toward warmer temperatures was unexpected for an Antarctic stenotherm and may indicate that thermal sensitivity of their mitochondria is not the driving force behind their stenothermy

Mitochondrial Energetics of Benthic and Pelagic Antarctic Teleosts

Antarctic fauna are highly adapted to the frigid waters of the Southern Ocean. This study describes the in vitro temperature sensitivity of oxygen consumption rates measured in liver mitochondria from the pelagic notothenioid Pleuragramma antarcticum between 5 and 35 C. Oxygen fluxes were measured after the addition of millimolar levels of pyruvate, malate, succinate and glutamate (state II, LEAK) and saturating levels of ADP [state III, oxidative phosphorylation (OXPHOS)]. State III respiration significantly decreased above 18.7 C. A comparison of the oxidative capacities among P. antarcticum and other notothenioids showed significant differences in state III respiration, where benthic species exhibited about 50 % lower rates than P. antarcticum . In addition, state III respiration rates normalized per milligram of mitochondrial protein of P. antarcticum were up to eight times higher than state III rates reported in the literature for other notothenioids. The comparatively high respiration rates measured in this study may be explained by our approach, which engaged both complexes I and II under conditions of oxidative phosphorylation. State III rates of independently activated complexes I and II were found to range from 42 to 100 % of rates obtained when both complexes were activated simultaneously in the same species. The remarkable tolerance of P. antarcticum OXPHOS toward warmer temperatures was unexpected for an Antarctic stenotherm and may indicate that thermal sensitivity of their mitochondria is not the driving force behind their stenothermy

Effects of Osmotic Stress on Oxygen Consumption of Drosophila Cells (Kc167)

This article investigates the effect of osmotic stress on a drosophila cell line called Kc167. The embryonic-derived fly (Drosophila melanogaster) cell line, Kc-167, was employed as a model for water-stress sensitivity in Arthropods. Like mammalian cells, cells derived from the fruit fly contain the same basic set of membranous components found in all eukaryotic cells. A series of experiments were conducted to characterize the mitochondrial repones of Kc167 cells to water stress. Precisely, the oxygen flux in a sealed respirometer chamber containing Kc167 cells was measured under hyperosmotic and control conditions. Mitochondrial uncouplers were used in some experiments for intact and chemically permeabilized cells to gain detailed information on mitochondrial integrity in response to increased solute concentration. Mitochondria are the primary ATP producer in the cell and consume oxygen in a process termed oxidative phosphorylation. Therefore, oxygen consumption rates can be used to assess the impact of water-limited states on cellular bioenergetics. Conducted experiments were performed to measure the following: • The basal oxygen consumption rates of Kc167 cells. • Consumption under conditions of oxidative stress • The oxygen consumption of chemically permeabilized cells • The max mitochondrial uncoupling that the Kc167 could withstand

Effects of Osmotic Stress on DNA and Cell Viability in a Desiccation-Sensitive Cell Line

Kc167 is a widely used Drosophila cell line, known to be sensitive to the extreme water loss caused by desiccation. In order to characterize the effects of this desiccation-sensitivity on DNA and cell viability, a series of osmotic stressors of differing concentrations were introduced to the cell line. These cells were then imaged via the Cytation1 cell imaging machine using fluorescence microscopy. Specifically, cells were stained using the DAPI staining solution, a blue fluorescent DNA stain that binds strongly to A-T rich regions within the DNA, forming a fluorescent complex. As DAPI more readily enters the membrane and thereby stains dead cells, instances of apoptosis caused by osmotic stress on cells can be characterized by increasing intensity of fluorescence. Both sucrose and sodium chloride were used to simulate the water loss relevant to that of desiccation. This was done in concentrations of 100mM, 250mM, and 500mM for both sucrose and sodium chloride

Functional and Conformational Plasticity of an Animal Group 1 LEA Protein

Group 1 (Dur-19, PF00477, LEA_5) Late Embryogenesis Abundant (LEA) proteins are present in organisms from all three domains of life, Archaea, Bacteria, and Eukarya. Surprisingly, Artemia is the only genus known to include animals that express group 1 LEA proteins in their desiccation-tolerant life-history stages. Bioinformatics analysis of circular dichroism data indicates that the group 1 LEA protein AfLEA1 is surprisingly ordered in the hydrated state and undergoes during desiccation one of the most pronounced disorder-to-order transitions described for LEA proteins from A. franciscana. The secondary structure in the hydrated state is dominated by random coils (42%) and β-sheets (35%) but converts to predominately α-helices (85%) when desiccated. Interestingly, AfLEA1 interacts with other proteins and nucleic acids, and RNA promotes liquid–liquid phase separation (LLPS) of the protein from the solvent during dehydration in vitro. Furthermore, AfLEA1 protects the enzyme lactate dehydrogenase (LDH) during desiccation but does not aid in restoring LDH activity after desiccation-induced inactivation. Ectopically expressed in D. melanogaster Kc167 cells, AfLEA1 localizes predominantly to the cytosol and increases the cytosolic viscosity during desiccation compared to untransfected control cells. Furthermore, the protein formed small biomolecular condensates in the cytoplasm of about 38% of Kc167 cells. These findings provide additional evidence for the hypothesis that the formation of biomolecular condensates to promote water stress tolerance during anhydrobiosis may be a shared feature across several groups of LEA proteins that display LLPS behaviors