How does the cold stenothermal gadoid Lota lota survive high water temperatures during summer?

The cold-stenothermal freshwater gadid Lota lota inhabiting the potamic regions of lowland rivers in central Europe, is exposed to summer temperatures up to 25 degrees C, which is far above the thermal preferendum of this species. Oxygen consumption rates, determined in field catches sampled at different times of the year, revealed that the basal metabolic rate is depressed during summer when water temperatures are high (152+/-16 micromol O2 100 g(-1) h(-1)at 22 degrees C in July compared to 250+/-33 micromol O2 100 g(-1) h(-1) at 6 degrees C in November). This observation led us to investigate whether the observed depression of the metabolic rate is caused by oxygen limitation due to thermal impairment of the ventilatory system, as has been observed in other species. Determination of anaerobic end products (lactate and succinate) in the liver tissue of fish caught at different sampling dates did not show an accumulation of anaerobic end products during the summer, indicating no oxygen limitation. Measurements of enzyme activities in the white musculature and liver suggest that enzymes involved in aerobic metabolism were down-regulated during summer, which may have contributed to the observed reduction of metabolic rate.

In vitro protein synthesis capacities in a cold stenothermal and a temperate eurythermal pectinid.

The translational system was isolated from the gills of the Antarctic scallop Adamussium colbecki (Smith) and the European scallop Aequipecten opercularis (Linnaeus) for in vitro protein synthesis capacities microg protein mg FW(-1) day(-1)) and the translational capacities of RNA (k(RNA in vitro) mg protein mg RNA(-1) day(-1)). In vitro protein synthesis capacity in the cold-adapted pectinid at 0 degrees C was similar to the one found in the temperate scallop at 25 degrees C. These findings might reflect cold compensated rates in Adamussium colbecki, partly explainable by high tissue levels of RNA. Cold-compensated in vitro protein synthesis capacities may further result from increments in the translational capacity of RNA. The thermal sensitivity of the translation machinery was slightly different in the two species, with significantly lower levels of Arrhenius activation energies E(a) and Q(10) in Adamussium colbecki in the temperature range 0-15 degrees C. Reduced protein synthesis and translational capacities were found in vitro in gills of long-term aquarium-maintained Adamussium colbecki and were accounted for by a loss of protein synthesis machinery, i.e. a reduction in RNA levels, as well as a decrease in the amount of protein synthesized per milligram of RNA (RNA translational capacity, k(RNA in vitro)). Such changes may involve food uptake or mirror metabolic depression strategies, like those occurring during winter. Consequences of high in vitro RNA translational capacities found in the permanently cold-adapted species are discussed in the context of seasonal food availability and growth rates at high latitudes.

The effect of fasting and refeeding on temperature preference, activity and growth of roach, Rutilus rutilus.

Most fish species are regularly subjected to periods of starvation during which a reduction of energy turnover might be favourable for the animal. This reduction of energy flux may be achieved by changes in thermal behaviour and/or swimming activity. We investigated such behavioural changes during starvation and subsequent refeeding in roach, Rutilus rutilus, with respect to energetic benefits and growth maximisation. Roach, acclimated to a wide range of temperatures (4, 12, 20, 24, 27 and 30 °C), were fed to excess, subjected to 3 weeks of starvation and subsequently refed in order to determine the temperature dependence of feeding rates, growth rates and conversion efficiency (K1) under control conditions and during compensatory growth. When exposed to a thermal gradient, control animals preferentially selected a temperature of 26.8±0.9 °C, which is in the range of the optimal temperatures for feeding, growth and conversion efficiency. Starving fish showed a distinct circadian pattern of the mean selected temperature (MST). They migrated to cooler water in the dark (MSTdark=22.8±1.1 °C) but returned to warmer water during daytime. This behaviour may be regarded as a trade-off between the potentially higher food density in warmer water areas and the energetic benefit of selecting cooler water patches. The circadian pattern of MST was gradually abandoned upon refeeding and control values were reached again after 3 weeks. Energetically more effective than behavioural hypothermia was the reduction of swimming activity. During starvation, activity peaks were slightly lower than under control conditions and mean daily activity decreased by about 50%. Swimming velocity, however, was not affected by feeding regime. After a period of starvation fish showed compensatory growth at all temperatures, even below 12 °C, where these animals normally do not grow. This suggests that after a period of starvation the critical temperature for growth shifts to lower values.

Physiological disturbances at critically high temperatures: a comparison between stenothermal antarctic and eurythermal temperate eelpouts (Zoarcidae)

The effect of gradually increased water temperature on the metabolism of temperate eelpout from the North Sea (Zoarces viviparus) and Antarctic eelpout (Pachycara brachycephalum) was investigated. Standard metabolic rate (SMR) was similar in cold-adapted P. brachycephalum and cold-acclimated Z. viviparus in the low temperature range. This indicates that Antarctic eelpout show no metabolic cold adaptation (as originally defined by Wohlschlag); however, they do show a compensatory increase of oxygen consumption compared to warm-acclimated eelpout. SMR increased more strongly with rising temperature in P. brachycephalum than in Z. viviparus, which is reflected in a higher Arrhenius activation energy for oxygen consumption (99+/-5 kJ mol(-)(1), versus 55+/-3 kJ mol(-)(1) for cold-acclimated Z. viviparus; means +/- s.d.). The intracellular pH in the white musculature of Z. viviparus follows alphastat regulation over the whole investigated temperature range and dropped at a rate of -0.016 pH units per degrees C between 3 degrees C and 24 degrees C. In Antarctic eelpout white muscle pH declined at a rate of -0.015 pH units per degrees C between 0 degrees C and 3 degrees C, but deviated from alphastat at higher temperatures, indicating that thermal stress leads to acid-base disturbances in this species. The upper critical temperature limit (Tc(II); characterised by a transition to anaerobic metabolism) was found to be between 21 degrees C and 24 degrees C for Z. viviparus and around 9 degrees C for P. brachycephalum. In both species a rise of succinate concentration in the liver tissue turned out to be the most useful indicator of Tc(II). Obviously, liver is more sensitive to heat stress than is white muscle. Accordingly, the energy status of white muscle is not diminished at Tc(II). Heat-induced hyperglycaemia was observed in Antarctic eelpout (at 9 degrees C and 10 degrees C), but not in common eelpout. Based on our results and on literature data, impaired respiration in combination with circulatory failure is suggested as the final cause of heat death. Our data suggest that the southern distribution limit of Zoarces viviparus is correlated with the limit of thermal tolerance. Therefore, it can be anticipated that global warming would cause a shift in the distribution of this species.

Temperature-dependent expression of cytochrome-c oxidase in Antarctic and temperate fish.

Seasonal acclimation versus permanent adaptation to low temperatures leads to a differential response in the expression of cytochrome-c oxidase (CCO) in temperate and Antarctic eelpouts. Although eurythermal eelpout from the North Sea (Zoarces viviparus) displayed a cold-induced rise of CCO activity in white muscle, enzyme activity in the cold stenothermal Antarctic eelpout Pachycara brachycephalum failed to reflect such a compensatory increase. In Antarctic eelpout, CCO activity correlates with transcript levels of mitochondrial encoded subunits of CCO (CCO I and CCO II), whereas cold-acclimated eelpout from the North Sea show lower enzyme activities than expected on the basis of mitochondrial mRNA levels. In these animals, CCO expression at low temperatures may be limited either by nuclear CCO transcripts or by posttranscriptional processes. These may comprise translation of the subunits or assembly of the CCO holoenzyme. mRNA levels of CCO IV, one of the nuclear encoded subunits, increased strongly during cold acclimation, indicating that the expression of CCO is likely not message limited in cold-acclimated Z. viviparus. Our data suggest that seasonal cold acclimation of Z. viviparus results in a modification of the relationship between transcription and translation or posttranslational processes. In permanently cold-adapted P. brachycephalum, on the other hand, CCO expression shows similar characteristics as in the warm-acclimated confamilial species, e.g., low levels of enzyme activity correlated with low levels of mitochondrial message.

High-energy turnover at low temperatures: recovery from exhaustive exercise in Antarctic and temperate eelpouts.

Earlier work on Notothenioids led to the hypothesis that a reduced glycolytic capacity is a general adaptation to low temperatures in Antarctic fish. In our study this hypothesis was reinvestigated by comparing changes in the metabolic status of the white musculature in two related zoarcid species, the stenothermal Antarctic eelpout Pachycara brachycephalum and the eurythermal Zoarces viviparus during exercise and subsequent recovery at 0 degreesC. In both species, strenuous exercise caused a similar increase in white muscle lactate, a drop in intracellular pH (pHi) by about 0.5 pH units, and a 90% depletion of phosphocreatine. This is the first study on Antarctic fish that shows an increase in white muscle lactate concentrations. Thus the hypothesis that a reduced importance of the glycolytic pathway is characteristic for cold-adapted polar fish cannot hold. The recovery process, especially the clearance of white muscle lactate, is significantly faster in the Antarctic than in temperate eelpout. Based on metabolite data, we calculated that during the first hour of recovery aerobic metabolism is increased 6.6-fold compared with resting rates in P. brachycephalum vs. an only 2.9-fold increase in Z. viviparus. This strong stimulation of aerobic metabolism despite low temperatures may be caused by a pronounced increase of free ADP levels, in the context of higher levels of pHi and ATP, which is observed in the Antarctic species. Although basal metabolic rates are identical in both species, the comparison of metabolic rates during situations of high-energy turnover reveals that the stenothermal P. brachycephalum shows a higher degree of metabolic cold compensation than the eurythermal Z. viviparus. Muscular fatigue after escape swimming may be caused by a drop of the free energy change of ATP hydrolysis, which is shown to fall below critical levels for cellular ATPases in exhausted animals of both species.

Temperature-dependent shift of pHi in fish white muscle: contributions of passive and active processes.

This study was designed to determine the mechanisms causing temperature-induced pH shifts in the white muscle of the marine teleost Zoarces viviparus. The white musculature undergoes an intracellular acidification with increasing body temperature at a slope of the pH-temperature relationship equal to -0.016 +/- 0.003 U/degree C. This is in good accordance with the overall relationship between the change in pK and the change in temperature of the intracellular proteins, which was determined to be -0.013 +/- 0.001 U/degree C. Thus the dissociation state of muscle proteins is kept fairly constant in white muscle of Zoarces viviparus. The passive component of the observed pH shift, which is due to the physicochemical response of the intracellular buffers to temperature change, accounts for only 35% of the pH transition. Ventilatory adjustment of intracellular PCO2 does not contribute to the temperature-induced shift of intracellular pH (pHi) in Zoarces viviparus. Therefore, the remaining 65% of pH adjustment must be ascribed to ion exchange mechanisms. The nonbicarbonate buffer value amounted to 34.4 +/- 2.3 meq.pH-1 kg cell water-1 at 12 degrees C and decreased slightly but not significantly with temperature. On the basis of our data we calculated that a removal of 0.52 mmol base equivalents.kg cell water-1.degree C-1 was necessary to shift pHi to its new steady state.

Profiles of nuclear and mitochondrial encoded mRNAs in developing and quiescent embryos of Artemia franciscana.

Embryos of the brine shrimp Artemia franciscana are able to withstand long bouts of environmental anoxia by entering a quiescent state during which metabolism is greatly depressed. Recent evidence supports a global arrest of protein synthesis during quiescence. In this study we measured the amounts of mRNA for a mitochondrial-encoded subunit of cytochrome c oxidase (COX I) and for nuclear-encoded actin during aerobic development, anaerobiosis, and aerobic acidosis (artificial quiescence imposed by intracellular acidification under aerobic conditions). The levels of both COX I and actin transcripts increased significantly during aerobic development. COX I mRNA levels were tightly correlated with previous measures of COX catalytic activity, which suggests that COX synthesis could be regulated by message concentration during aerobic development. The ontogenetic increase for these mRNAs was blocked by anoxia and aerobic acidosis. Importantly, the levels of COX I and actin mRNA did not decline appreciably during the 6 h bouts of quiescence, even though protein synthesis is acutely arrested by these same treatments. Thus, the constancy of mRNA levels during quiescence indicate that reduced protein synthesis is not caused by message limitation, but rather, is likely controlled at the translational level. One advantage of this regulatory mechanism is the conservation of mRNA molecules during quiescence, which would potentially favor a quick resumption of translation as soon as oxygen is returned to the embryos. Finally, because anoxia and aerobic acidosis are both characterized by acidic intracellular pH, the reduction in pH may serve, directly or indirectly, as one signal regulating levels of mRNA in this embryo during quiescence.

Downregulation of cellular metabolism during environmental stress: mechanisms and implications.

Survival time of organisms during exposure to environmental stresses that limit energy availability is, in general, directly related to the degree of metabolic depression achieved. The energetic cost savings realized by the organism is a consequence primarily of the ability to depress ion pumping activities of cells, macromolecular synthesis, and macromolecular turnover. Evidence supporting the concept of channel arrest-the reduction in ion leakage across cell membranes during hypometabolic states-has highlighted the energetic benefits of limiting ATP turnover related to cellular ion homeostasis. Depression of protein synthesis results in substantial bioenergetic savings. However, when protein synthesis is arrested, the preservation of macromolecules becomes increasingly important as the duration of quiescence is extended because the cellular capacity for replenishing these components is reduced. It is likely that the rate of macromolecular degradation is a key feature that sets the upper time limit for survival during chronic environmental stress.