A re-interpretation of the effect of temperature on the metabolism of certain marine invertebrates

Measurements by nieans of an oxygen electrode of the rate of oxygen consumption of a wide variety of common intertidal invertebrates show that at least two rates of uptake can be distinguished in the intact animal. First, there is a rapid rate of oxygen uptake corresponding with activity and second there is a slower rate which corresponds with periods of quiescence and which alternates with the fast rate. Intermediate rates correspond with intermediate rates of overt activity. The maximal and minimal rates of oxygen uptake are affected differently by temperature change. The fast rate corresponding to activity ("active metabolism") increases with temperature in approximate agreement with Arrhenius's law. The slow rate corresponding to quiescence ("maintenance metabolism") does not vary with temperature over much of the range 7 to 22.5°C. It is concluded that, contrary to common belief, the intertidal invertebrates studied have a "basal metabolic rate" with a Q₁₀ of approximately one over much of the normal environmental temperature range and in this respect are well suited to life in a habitat where rapid fluctuations in temperature occur.     

Temperature and Food as Factors Influencing Oxygen Consumption of Intertidal Organisms, Particularly Limpets

SYNOPSIS: In intertidal animals rates of oxygen consumptionspan an order of magnitude (even when standardised for bodysize and temperature). This may reflect different adaptationsto this variable habitat. Temperatures range widely and potentiallyincrease metabolic expenditure during the heat of the day, whileperiods of feeding may be restricted. The balance between intakeand expenditure of energy may limit zonation, body size, andeven mode of reproduction. One pattern is that species sufferingshortage of food ("conservers") have low rates of oxygen consumptionwhile those with abundant food ("exploiters") have high rates.Conservers appear to have several means of reducing metaboliccosts, but exploiters maintain high (seemingly wasteful) ratesof oxygen consumption. Oxygen consumption cannot, however, be considered separately,since it reflects turnover,including growth rate and reproductiveoutput. There is also the question whether differences betweenspecies are genetically controlled, or simply phenotypic responsesto food availability. Comparisons of populations with differentamounts of food suggest that exploiters substantially reduceoxygen consumption when food is scarce, but conservers are relativelyfixed in their pattern of slow growth and low reproductive output,even if food is experimentally increased. Growth and reproduction are positively correlated, at leastwithin taxonomically related groups (as exemplified by patellaceanlimpets) but both are inversely related to longevity.Thus growthand reproduction are not necessarily "traded off," but are bothlow in "conservers" and both high in "exploiters."     

Effect of Fluctuations in Temperature on the Metabolism of Intertidal Invertebrates

SVNOPSIS. Compensatory changes in the level of activity of intertidalorganisms may occur in response to the thermal conditions whichprevail in the habitat. These involve an increase in the activityof those animals subjected to low temperatures and a correspondingsuppression of activity in those animals subjected to high temperaturesso that organisms with a wide geographical range, or livingover a range of shore levels, have comparable rates of activity.However, the rate of activity varies markedly with short-termfluctuations in temperature, as does the rate of respirationof active animals. Recent evidence suggests that the rate ofrespiration of quiescent animals is relatively independent oftemperature over the normal environmental range, and thus themetabolism is well-suited to an environment where rapid fluctuationsin temperature occur. Further, the extent of the thermal rangeover which metabolism is relatively independent of temperatureis modifiable according to season and storage-temperature. Similarchanges occur in the respiration of cell-free homogenates ofcertain intertidal organisms. Acclimation in such organismsinvolves not only a modification in the level of the activeand standard rates of metabolism but also an alteration in theform of the rate/temperature curve such that the range of temperature-independentmetabolism is appropriate to the thermal conditions prevailingin the habitat.     

Oxidative activity of poikilotherm mitochondria as a function of temperature

In Newell & Northcroft (1967) evidence was presented which showed that the metabolism of certain intertidal invertebrates at rest does not vary greatly with temperature change. The following work was undertaken to determine whether temperature-independent metabolism could be demonstrated in suspensions of mitochondria extracted from poikilotherm tissue and whether it could be inferred that there was any difference in the ability of poikilotherms from different habitats to adjust to short-term temperature change.
It was found that the rate of reaction/temperature curve of mitochondria isolated from the skate Raia had a flattened region below 10C but above this the rate of oxidation of succinate and pyruvate increased regularly with temperature. The curve for intertidal forms had a gradual slope extending as high as 20C in the mussel Mytilus edulis while in the terrestrial snail Helix aspersa the gradual slope reached 27.5C after which the rate of oxidation of substrate increased sharply before declining towards higher temperatures. Finally, the curve for mitochondria isolated from the desert locust Schistocerca gregaria showed a gradual slope up to 35C after which a sharp increase in the rate of oxidation of substrate occurred. Thus the extent of the gradual slope corresponds with the normal environmental temperature range to which the tissues of the poikilotherms are subjected and the general form of the R-T curve is such that rapid fluctuations within the normal environmental temperature range have relatively little effect on the rate of mitochondrial activity.     

Oxidative activity of poikilotherm mitochondria as a function of temperature

In Newell & Northcroft (1967) evidence was presented which showed that the metabolism of certain intertidal invertebrates at rest does not vary greatly with temperature change. The following work was undertaken to determine whether temperature-independent metabolism could be demonstrated in suspensions of mitochondria extracted from poikilotherm tissue and whether it could be inferred that there was any difference in the ability of poikilotherms from different habitats to adjust toshort-term temperature change.
It was found that the rate of reaction/temperature curve of mitochondria isolated from the skate Raia had a flattened region below 10°C but above this the rate of oxidation of succinate and pyruvate increased regularly with temperature. The curve for intertidal forms had a gradual slope extending as high as 20°C in the mussel Mytilus edulis while in the terrestrial snail Helix aspersa the gradual slope reached 27.5°C after which the rate of oxidation of substrate increased sharply before declining towards higher temperatures. Finally, the curve for mitochondria isolated from the desert locust Schistocerca gregaria showed a gradual slope up to 35°C after which a sharp increase in the rate of oxidation of substrate occurred. Thus the extent of the gradual slope corresponds with the normal environmental temperature range to which the tissues of the poikilotherms are subjected and the general form of the R-T curve is such that rapid fluctuations within the normal environmental temperature range have relatively little effect on the rate of mitochondrial activity.     

Temperature influences whole-animal rates of metabolism but not protein synthesis in a temperate intertidal isopod

The effects of temperature on two important biological rate processes, whole-body rates of oxygen uptake (M dot o2) and protein synthesis (k(s)), were investigated in the temperate intertidal isopod Ligia oceanica at two different times of the year. Animals were collected in January (winter) and June (summer) and either subjected to an acute temperature change after 24 h (acclimatized) or acclimated to various temperatures for 4 wk. In both cases, M dot o2 increased with temperature, with a Q(10) of 2.2 between 5 degrees and 20 degrees C, but increased in thermal sensitivity at 25 degrees C. Winter isopods were characterized by significantly higher M dot o2 levels, greater thermal sensitivities, and lower thermal tolerances than summer animals. Seasonal differences in M dot o2 persisted after acclimation, indicating that temperature alone was not responsible for the changes. In sharp contrast, whole-body k(s) showed no variation with temperature, although overall rates decreased upon acclimation. In acclimatized animals, k(s) was higher in the summer than in the winter. After acclimation, a compensatory increase in RNA capacity in winter animals reversed this situation. The temperature independence of whole-body k(s) in L. oceanica could ensure survival in a highly liable thermal environment, as thermal tolerances of intertidal invertebrates are thought to be more closely related to protein than to energy metabolism.     

Effect of transfer of fragment of chromosome 13, containing gene i16st on parameters of mouse temperature homeostasis

Mechanisms of maintenance of temperature homeostasis in warm and under effect of cold were studied in mice of AKR strain and of its coherent strain AKR.CBA-D13Mit76 with the changed gene i16st encoding the gp 130 receptor, via which IL-6 performs its action. Under thermoneutral conditions and under action of cold, there were recorded temperature parameters, total oxygen consumption, carbon dioxide release, respiratory coefficient, and electrical muscle activity. Animals of the studied strains demonstrated different reactions to equal cold effect. At cooling, all mice of the AKR strain entered the state of hypothermia by decreasing metabolism. Mice of the AKR.CBA-D13Mit76 line showed 2 different types of reaction: 39 % of the animals of this strain reacted like mice of the AKR strain, but the majority (61 %) resisted actively to the cold action, which was manifested as a marked increase of metabolism. Taking into account the gene penetrance, this can indicate effect of the gene i16st on choice of the active ("regulated") or passive ("dependent") way of the organism reacting to temperature actions.     

Superweak irradiation of marine invertebrates

The inhibitory analysis of the spontaneous ("ultraweak") and the luminole-induced chemiluminescence of marine Sycon sponges and Aiptasia actinias supports the idea that ultraweak photon emission of marine invertebrates is a consequence of Ca(2+)-dependent processes related to the interaction of reactive oxygen species with some endogenous fluorophore substrates.     

Factors Affecting the Respiration of Intertidal Invertebrates

The nature and rate of gas exchange in intertidal organismsis variable and is the product of an extremely complex environmentalsituation. Such influences may be grouped into tidal-dependentfactors such as the proportion of time spent exposed to air,the magnitude of environmental temperature fluctuations, andthe availability of food. Factors which may be regarded as generallyoperating independently of tidal level include latitudinal variationsin environmental temperature. Superimposed upon such spatialenvironmental variations are seasonal factors such as temperatureand photoperiod, which impose a temporal cycle upon the metabolicrate. Finally, there are numerous endogenous factors such asbody size, activity level, and stage of development which profoundlyinfluence the rate of respiration. It can be shown that somefactors, such as aerial or aquatic conditions, may affect respirationqualitatively. Under aquatic conditions gas exchange is withthe surrounding sea water, but with increasing exposure to airintertidal invertebrates are able to respire aerially and towithstand the water loss associated with this process. Manyare also able to respire anaerobically under conditions of stress.In contrast, factors such as activity level, body size, nutritionalconditions, and exposure temperature affect respiration quantitativelyand are often interdependent. The metabolism of intertidal animalsis then endowed with considerable flexibility between the extremesset by the active and standard rates of respiration. This activerate of respiration is markedly temperature-dependent in mostinstances, whereas the standard rate, which is characteristicof quiescent animals, is not only lower but often has a lowtemperature coefficient. A reduction in respiration conservesmetabolic reserves during periods of stress, and the low temperaturecoefficient further minimizes depletion of such reserves despitethe high environmental temperatures which often prevail duringthe intertidal period.     

A mutation in subunit I of cytochrome oxidase from Rhodobacter sphaeroides results in an increase in steady-state activity but completely eliminates proton pumping

The heme-copper oxidases convert the free energy liberated in the reduction of O(2) to water into a transmembrane proton electrochemical potential (protonmotive force). One of the essential structural elements of the enzyme is the D-channel, which is thought to be the input pathway, both for protons which go to form H(2)O ("chemical protons") and for protons that get translocated across the lipid membrane ("pumped protons"). The D-channel contains a chain of water molecules extending about 25 A from an aspartic acid (D132 in the Rhodobacter sphaeroides oxidase) near the cytoplasmic ("inside") enzyme surface to a glutamic acid (E286) in the protein interior. Mutations in which either of these acidic residues is replaced by their corresponding amides (D132N or E286Q) result in severe inhibition of enzyme activity. In the current work, an asparagine located in the D-channel has been replaced by the corresponding acid (N139 to D; N98 in bovine enzyme) with dramatic consequences. The N139D mutation not only completely eliminates proton pumping but, at the same time, confers a substantial increase (150-300%) in the steady-state cytochrome oxidase activity. The N139D mutant of the R. sphaeroides oxidase was further characterized by examining the rates of individual steps in the catalytic cycle. Under anaerobic conditions, the rate of reduction of heme a(3) in the fully oxidized enzyme, prior to the reaction with O(2), is identical to that of the wild-type oxidase and is not accelerated. However, the rate of reaction of the fully reduced enzyme with O(2) is accelerated by the N139D mutation, as shown by a more rapid F --> O transition. Whereas the rates of formation and decay of the oxygenated intermediates are altered, the nature of the oxygenated intermediates is not perturbed by the N139D mutation.     

Relative contributions of "pressure pump" and "peristaltic pump" to gastric emptying

The relative contributions to gastric emptying from common cavity antroduodenal pressure difference ("pressure pump") vs. propagating high-pressure waves in the distal antrum ("peristaltic pump") were analyzed in humans by high-resolution manometry concurrently with time-resolved three-dimensional magnetic resonance imaging during intraduodenal nutrient infusion at 2 kcal/min. Gastric volume, space-time pressure, and contraction wave histories in the antropyloroduodenal region were measured in seven healthy subjects. The subjects fell into two distinct groups with an order of magnitude difference in levels of antral pressure activity. However, there was no significant difference in average rate of gastric emptying between the two groups. Antral pressure history was separated into "propagating high-pressure events" (HPE), "nonpropagating HPEs," and "quiescent periods." Quiescent periods dominated, and average pressure during quiescent periods remained unchanged with decreasing gastric volume, suggesting that common cavity pressure levels were maintained by increasing wall muscle tone with decreasing volume. When propagating HPEs moved to within 2-3 cm of the pylorus, pyloric resistance was found statistically to increase with decreasing distance between peristaltic waves and the pylorus. We conclude that transpyloric flow tends to be blocked when antral contraction waves are within a "zone of influence" proximal to the pylorus, suggesting physiological coordination between pyloric and antral contractile activity. We further conclude that gastric emptying of nutrient liquids is primarily through the "pressure pump" mechanism controlled by pyloric opening during periods of relative quiescence in antral contractile wave activity.     

Some properties of potassium-stimulated calcium influx in presynaptic nerve endings

Potassium-stimulated 45Ca entry into rat brain synaptosomes was measured at times ranging from 1 to 60 s. The K-rich solutions were used to depolarize the synaptosomes. Backflux of 45Ca from the synaptosomes was negligible during the first 10-20 s of incubation. An initial ("fast") phase of K-stimulated Ca entry, lasting from 1 to 2 s was observed. This phase was inhibited by low concentrations of La (KI approximately equal to 0.3 microM). It was also abolished ("inactivated") by incubating the synaptosomes in depolarizing solutions (containing veratridine, gramicidin, or elevated [K]o) before the addition of 45Ca. An additional long lasting ("slow") phase of K-stimulated Ca entry was also detected. This "slow" Ca entry was much less sensitive to La (KI > 100 microM) and was not affected by depolarizing the synaptosomes before the addition of 45Ca. The rate of influx during the fast phase was about four times the rate of Ca influx during the slow phase. Neither the fast nor slow phase of Ca entry was sensitive to tetrodotoxin (10 microM), a potent blocker of Na channels, but both phases were inhibited by Ni, Mn, Mg, and other agents that block Ca channels. The data are consistent with the presence of two distinct populations of voltage-regulated, divalent cation-selective pathways for Ca entry in presynaptic brain nerve endings.     

The environmental effects of salinity and temperature on the oxygen consumption and total body osmolality of the marine flatworm Procerodes littoralis

The present study examined the effect of salinity and temperature on the rate of oxygen consumption and total body osmolality of the triclad turbellarian Procerodes littoralis, a common marine flatworm normally found in areas where freshwater streams run out over intertidal areas. Extremes in environmental factors encountered by P. littoralis were recorded at the study site. These were salinity (0-44 psu), temperature (2.7-24.9 °C) and oxygen concentration (2.8-16.1 mg l⁻¹). Respirometry experiments showed minimal oxygen consumption rates at the salinity extremes encountered by the study species (0 and 40 psu). Further experiments showed relatively constant oxygen consumption rates over the temperature range 5-20 °C and elevated consumption rates at temperatures above 25 °C. Total body osmolality of P. littoralis increased with increasing salinity. The study illustrates how a marine flatworm uses integrated physiological and behavioural mechanisms to successfully inhabit an environment that is predominantly freshwater for up to 75% of the tidal cycle.     

Staying alive

Quiescence is a state of reversible cell cycle arrest that can grant protection against many environmental insults. In some systems, cellular quiescence is associated with a low metabolic state characterized by a decrease in glucose uptake and glycolysis, reduced translation rates and activation of autophagy as a means to provide nutrients for survival. For cells in multiple different quiescence model systems, including Saccharomyces cerevisiae, mammalian lymphocytes and hematopoietic stem cells, the PI3Kinase/TOR signaling pathway helps to integrate information about nutrient availability with cell growth rates. Quiescence signals often inactivate the TOR kinase, resulting in reduced cell growth and biosynthesis. However, quiescence is not always associated with reduced metabolism; it is also possible to achieve a state of cellular quiescence in which glucose uptake, glycolysis and flux through central carbon metabolism are not reduced. In this review, we compare and contrast the metabolic changes that occur with quiescence in different model systems.     

Staying alive: Metabolic adaptations to quiescence

Quiescence is a state of reversible cell cycle arrest that can grant protection against many environmental insults. In some systems, cellular quiescence is associated with a low metabolic state characterized by a decrease in glucose uptake and glycolysis, reduced translation rates and activation of autophagy as a means to provide nutrients for survival. For cells in multiple different quiescence model systems, including Saccharomyces cerevisiae, mammalian lymphocytes and hematopoietic stem cells, the PI3Kinase/TOR signaling pathway helps to integrate information about nutrient availability with cell growth rates. Quiescence signals often inactivate the TOR kinase, resulting in reduced cell growth and biosynthesis. However, quiescence is not always associated with reduced metabolism; it is also possible to achieve a state of cellular quiescence in which glucose uptake, glycolysis and flux through central carbon metabolism are not reduced. In this review, we compare and contrast the metabolic changes that occur with quiescence in different model systems.     

Comparative respiration of some tropical intertidal gastropods

The rate of oxygen uptake of three species of tropical intertidal gastropods,Nerita tessellata Gmelid., N. Versicolor Gmelin and N. Peloronta L., have been investigated under different environmental conditions. In all species the rates increased from 30 to 37°C, were depressed at 20 °C and were not size-dependent at that temperature. The rates of oxygen uptake varied between individuals collected from different habitats and were greater in areas of higher mean maximum daily temperatures, lower rainfall and lower wind speeds than in areas with the reverse conditions. A decrease in oxygen uptake was shown to occur with an increase of exposure time of the animals on the shore and in the laboratory. Rates were higher in two species collected at the time of high tide than those collected at the same level after exposure to low tides.     

Ammonium Assimilation and the Role of [gamma]-Aminobutyric Acid in pH Homeostasis in Carrot Cell Suspensions

In vivo 15N NMR spectroscopy was used to monitor the assimilation of ammonium by cell-suspension cultures of carrot (Daucus carota L. cv Chantenay). The cell suspensions were supplied with oxygen in the form of either pure oxygen ("oxygenated cells") or air ("aerated cells"). In contrast to oxygenated cells, in which ammonium assimilation had no effect on cytoplasmic pH, ammonium assimilation by aerated cells caused a decrease in cytoplasmic pH of almost 0.2 pH unit. This led to a change in nitrogen metabolism resulting in the accumulation of [gamma]-aminobutyric acid. The metabolic effect of the reduced oxygen supply under aerated conditions could be mimicked by artificially decreasing the cytoplasmic pH of oxygenated cells and was abolished by increasing the cytoplasmic pH of aerated cells. The activity of glutamate decarboxylase increased as the cytoplasmic pH declined and decreased as the pH recovered. These findings are consistent with a role for the decarboxylation of glutamate, a proton-consuming reaction, in the short-term regulation of cytoplasmic pH, and they demonstrate that cytoplasmic pH influences the pathways of intermediary nitrogen metabolism.     

Transport of 6-deoxyglucose in Saccharomyces cerevisiae.

The uptake of 6-deoxyglucose was measured in wild-type Saccharomyces cerevisiae, in a double mutant strain lacking activity for hexokinases A and B (hxkl hxk2), in a triple mutant strain lacking activity for both hexokinases and glucokinase (hxkl hxk2 glk), and in the triple mutant with high levels of activity of single kinases restored by introduction of the cloned genes. In the wild-type strain, uptake of the glucose analog showed two components, with Km values of ca. 20 mM ("high affinity") and 250 mM ("low affinity"), respectively. The double mutant also had high- and low-affinity components, but the triple mutant showed only low-affinity uptake. Reintroduction of the single kinases to the triple mutant restored high-affinity uptake. (Other experiments on 6-deoxyglucose uptake are also presented, including the apparent use of the galactose transport system when induced.) These results show that the recent implication of the kinases in transport of glucose (L.F. Bisson and D.G. Fraenkel, Proc. Natl. Acad. Sci. U.S.A. 80:1730-1734, 1983) applies equally to the nonmetabolized analog 6-deoxyglucose and suggests that the role of the kinases in transport is not merely a consequence of metabolism of the transported compound.     

High-speed running performance is largely unaffected by hypoxic reductions in aerobic power.

We tested the importance of aerobic metabolism to human running speed directly by altering inspired oxygen concentrations and comparing the maximal speeds attained at different rates of oxygen uptake. Under both normoxic (20.93% O2) and hypoxic (13.00% O2) conditions, four fit adult men completed 15 all-out sprints lasting from 15 to 180 s as well as progressive, discontinuous treadmill tests to determine maximal oxygen uptake and the metabolic cost of steady-state running. Maximal aerobic power was lower by 30% (1.00 +/- 0.15 vs. 0.77 +/- 0.12 ml O2. kg-1. s-1) and sprinting rates of oxygen uptake by 12-25% under hypoxic vs. normoxic conditions while the metabolic cost of submaximal running was the same. Despite reductions in the aerobic energy available for sprinting under hypoxic conditions, our subjects were able to run just as fast for sprints of up to 60 s and nearly as fast for sprints of up to 120 s. This was possible because rates of anaerobic energy release, estimated from oxygen deficits, increased by as much as 18%, and thus compensated for the reductions in aerobic power. We conclude that maximal metabolic power outputs during sprinting are not limited by rates of anaerobic metabolism and that human speed is largely independent of aerobic power during all-out runs of 60 s or less.     

Effects of salinity and temperature on oxygen consumption in a freshwater population of Palaemonetes antennarius (Crustacea,decapoda)

• 1.1. After step-like increases in salinity the shrimps exhibit the smallest increase in oxygen consumption in the lower salinity range. At higher salinities the shrimps show longer recovery times and greater increases in the metabolic rate after salinity shock.
• 2.2. In steady-state experiments, the shrimps display the lowest oxygen consumption rates near the isosmotic point. The lowest metabolic rates occur at salinities of 3‰ and 10‰ At salinities of 20‰ and above the rate of metabolism increases by 20–30%.
• 3.3. The calculated osmoregulatory work for animals in fresh water amounts to only 2.7% of routine metabolism and drops to 1.1% for shrimps in 3‰ and 0.7% in 5‰ salinity.
• 4.4. Locomotory activity in the form of position change was not responsible for the increased oxygen consumption of the animals after salinity shocks. A “tentative swimming activity” by fast and frequent beating of the pleopods without position change may be an important factor in the increase of metabolic rates.
• 5.5. In its temperature response, the brackish water population has a higher metabolic rate than the freshwater one. Between 5 and 35°C Q ₁₀-values range from 4.01 to 1.37.