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Adaptative features of ectothermic enzymes--II. The effects of acclimation temperature on the malate dehydrogenase of the spot. Leiostomus xanthurus.

1. Isozyme patterns and thermostability of the skeletal muscle and heart malate dehydrogenase (s-MDH) from the spot acclimated to different temperatures were examined. 2. No changes in isozyme patterns were seen for MDH in any of the tissues examined in response to 15 degrees and 20 degrees C acclimation. 3. The A-homodimer, which was more thermostable showed an increase in its relative activity, whereas the B-homodimer, which was more heat sensitive, showed a decrease during warm acclimation. However, after different periods of low temperature incubation, these samples showed a decrease in their subunit ratios. 4. Concerning the effect to thermal acclimation on the thermostability of MDH, it was found that skeletal muscle samples of the 30 degrees C-acclimated spot were more stable to heat than the 20 degrees and 15 degrees C-acclimated fishes.

Acclimatization↗

Increased respiration in skeletal muscle mitochondria from cold-acclimated ducklings: uncoupling effects of free fatty acids.

Intermyofibrillar mitochondria from skeletal muscle (m. gastrocnemius) and liver mitochondria were isolated from cold-acclimated (4 degrees C) or control (30 degrees C) 4-week old ducklings. The respiratory rate of isolated mitochondria, with Na-succinate as substrate, was followed polarographically at 25 degrees C in order to determine the basal respiratory rate, the rate of respiration in the presence of free fatty acids (FFA) (Na-palmitate), and the fully uncoupled rate, after addition of FCCP. The basal respiration (which in liver mitochondria was unaffected by acclimation to cold) was higher (+53%) in intermyofibrillar mitochondria from cold-acclimated ducklings than from controls, and the maximal FCCP-stimulated respiration was also increased (+98%) by acclimation to cold. FFA-stimulated respiration increased as a function of FFA concentration in both types of mitochondria. The increase in respiration due to FFA was about double in intermyofibrillar mitochondria from cold-acclimated ducklings than that of controls, but in liver mitochondria there was no increase due to cold. The membrane potential was estimated by the dye safranine in the absence or in the presence of FFA in the incubation medium. There were no significant differences in the basal membrane potential in the two groups and the addition of FFA led to the same depolarization in both groups. The significance of these alterations for acclimation to cold is discussed.

Acclimatization↗

Cold acclimation and oxygen consumption in the thymus.

Mitochondrial uncoupling protein 1 is usually associated with brown adipose tissue but has recently been discovered in rat and mouse thymus. We wished to establish whether there was a thermogenic role for UCP 1 in thymus and thus examined the effect of 5 weeks cold-acclimation on rat thymus tissue abundance, thymocyte oxygen consumption, thymus mitochondrial abundance, uncoupling protein 1 expression and function. We found that thymocytes from cold-acclimated rats had oxygen consumption rates 8 times less than those from rats held at room temperature and that thymocytes from cold-acclimated rats or rats kept at room temperature were noradrenaline insensitive. In addition, we found that thymus tissue or mitochondrial abundance was not increased after cold-acclimation. However uncoupling protein 1 expression per unit mass of mitochondria was increased after cold-acclimation, as determined by immunoblotting (approximately 1.7-fold) and GDP binding (approximately 1.5-fold). Consistent with our protein expression data, we also observed an increased, state 4 (approximately 1.5-fold), GDP-inhibitable (approximately 1.3-fold) and palmitate activatable (approximately 1.6-fold) oxygen consumption rates in isolated thymus mitochondria. However, extrapolation of our data showed that cold-acclimation only increased the amount of UCP 1 per gram of thymus tissue approximately 1.2-fold. Taken together, we conclude that UCP 1 does not have a thermogenic role in thymus.

Acclimatization↗

Effect of temperature acclimation on metabolism and hemocyanin binding affinities in two crayfish, Procambarus clarkii and Procambarus zonangulus.

Procambarus clarkii and Procambarus zonangulus are two of the most widespread crayfish species in North America. In regions where their ranges overlap species composition can vary greatly. The physiological basis for this variable species composition is unknown. Temperature and oxygen level are two parameters that vary in shallow water habitats. We examined the metabolic rate and hemocyanin binding affinities in relation to thermal history. Temperature acclimation did not have the predicted effect on metabolic rate. Acclimation to high temperature (30 degrees C) decreased metabolic rate at 35 degrees C for both species. Low temperature acclimation (10 degrees C) resulted in 20% mortality in P. clarkii and 100% mortality in P. zonangulus when exposed to 35 degrees C. The range of P. clarkii is known to extend farther south than that of P. zonangulus, and this response may be a consequence of adaptations to higher temperatures in this range. Hemocyanin binding affinity was directly affected by assay and acclimation temperature. The highest P(50) values were recorded for crayfish of both species acclimated to 10 degrees C and assayed at 30 degrees C. There was also a shift in the isoelectric points of hemolymph proteins (possibly due to structural changes) that correlated with and an increase in the hemocyanin binding affinity following acclimation to high temperatures (30 degrees C) in both species.

Acclimatization↗

Acclimation of Daphnia magna to environmentally realistic copper concentrations.

It may be hypothesised that as the bioavailable background concentration of an essential metal increases (within natural limits), the natural tolerance (to the metal) of the acclimated/adapted organisms and communities will increase. In this study the influence of acclimation to different copper concentrations on the sensitivity of the freshwater cladoceran Daphnia magna Straus was investigated. D. magna was acclimated over three generations to environmentally relevant copper concentrations ranging from 0.5 to 100 microg Cu/l (copper activity: 7.18 x 10(-15) to 3700 x 10(-12) M Cu2+). A modified standard test medium was used as culture and test medium. Medium modifications were: reduced hardness (lowered to 180 mg CaCO3/l) and addition of Aldrich humic acid at a concentration of 5 mg DOC/l (instead of EDTA). The effects of acclimation on these organisms were monitored using acute mortality assays and long-term assays in which life table parameters, copper body concentrations and energy reserves were used as test endpoints. Our results showed a two-fold increase in acute copper tolerance with increasing acclimation concentration for second and third generation organisms. Copper acclimation concentrations up to 35 microg Cu/l (80 pM Cu2+) did not affect the net reproduction and the intrinsic growth rate. The energy reserves of the acclimated daphnids revealed an Optimal Concentration range (OCEE) and concentrations between 5 and 12 microg Cu/l (0.5-4.1 pM Cu2+) and 1 and 35 microg Cu/l (0.023-80 pM Cu2+) seemed to be optimal for first and third generation daphnids, respectively. Lower and higher copper concentrations resulted in deficiency and toxicity responses. It was also demonstrated that up to 35 microg Cu/l, third generation daphnids were able to regulate their total copper body concentration. These results clearly indicate that bioavailable background copper concentrations present in culture media have to be considered in the evaluation of toxicity test results, especially when the toxicity data are used for water quality guideline derivation and/or ecological risk assessment for metals.

Adaptation, Physiological↗

Toxaphene detoxification and acclimation in Daphnia magna: do cytochrome P-450 enzymes play a role?

Toxaphene is a persistent environmental contaminant that has been shown to alter male production in Daphnia magna and to induce P-450 activity in mammals. Cytochrome P-450-mediated metabolism may lead to xenobiotic detoxification resulting in acclimation. To determine if D. magna acclimate to toxaphene via P-450 pathways, chronic and acute toxicity tests were conducted with D. magna exposed to toxaphene in the presence and absence of piperonyl butoxide (PBO), an inhibitor of cytochrome P-450 enzymes. Toxaphene exposure increased male production in acute but not chronic assays, indicating that D. magna may acclimate to chronic toxaphene exposure. Upon co-administration of toxaphene and PBO in chronic tests, D. magna exhibited a decline in growth rate, fecundity and survival. The observed toxaphene acclimation in chronic tests, along with its increased toxicity in the presence of a P-450 suppressor, suggests that P-450 enzymes may contribute to detoxification and subsequent acclimation of D. magna to chronic toxaphene exposure. Additional chronic toxicity tests indicated that toxaphene acclimation occurs between 7 and 12 days following initial exposure, at which time sex determination is no longer affected. Thus, sublethal toxaphene toxicity effects such as reproductive impairments may be detectable with acute but not chronic tests, potentially due to the upregulation of P-450 isozymes.

Acclimatization↗

The role of raffinose in the cold acclimation response of Arabidopsis thaliana.

In many plants raffinose family oligosaccharides are accumulated during cold acclimation. The contribution of raffinose accumulation to freezing tolerance is not clear. Here, we investigated whether synthesis of raffinose is an essential component for acquiring frost tolerance. We created transgenic lines of Arabidopsis thaliana accessions Columbia-0 and Cape Verde Islands constitutively overexpressing a galactinol synthase (GS) gene from cucumber. GS overexpressing lines contained up to 20 times as much raffinose as the respective wild-type under non-acclimated conditions and up to 2.3 times more after 14 days of cold acclimation at 4 degrees C. Furthermore, we used a mutant carrying a knockout of the endogenous raffinose synthase (RS) gene. Raffinose was completely absent in this mutant. However, neither the freezing tolerance of non-acclimated leaves, nor their ability to cold acclimate were influenced in the RS mutant or in the GS overexpressing lines. We conclude that raffinose is not essential for basic freezing tolerance or for cold acclimation of A. thaliana.

Acclimatization↗

Adjustments of the enzymatic complement for polyol biosynthesis and accumulation in diapausing cold-acclimated adults of Pyrrhocoris apterus.

The capacity to accumulate winter polyols (mainly ribitol and sorbitol) during cold-acclimation in Pyrrhocoris apterus is restricted only to the adults that have previously entered diapause. The enzymatic complement involved in polyol biosynthesis was found to differ in a complex manner between diapause and non-diapause adults. Nearly 100% of glycogen phosphorylase (GPase) was present in its active form in non-diapause adults irrespective of their acclimation status. In contrast, less than 40% of GPase was present in its active form in diapause adults prior to cold-acclimation and the inactive form was rapidly activated upon transition from 5 to 0 degrees C, concomitantly with the start of rapid polyol accumulation. The flow of carbon released by activation of glycogen degradation might be routed to the pentose cycle because the activity of glucose-6-P dehydrogenase (G(6)P-DH) was significantly higher and it increased with cold-acclimation in diapause adults while it was relatively low and it decreased with cold-acclimation in non-diapause adults. Reducing equivalents in the form of NADPH, which were generated in the pentose cycle, might require re-oxidation. Such re-oxidation might be achieved during reduction of sugars to polyols. The activity of NADP(H)-dependent aldose reductase (AR) was about 20-fold higher in diapause than in non-diapause adults. Similarly, the activity of NAD(H)-dependent polyol dehydrogenase (PDH) was higher in diapause adults. In addition, we found a very high activity of an unusual enzyme, NADP(H)-dependent ketose reductase (KR), exclusively in diapause adults. KR might be involved in reduction of fructose to sorbitol. Although its affinity for fructose as a substrate was low (K(M)=0.64M), its activity was about 10-fold higher than that of PDH with fructose. Moreover, the activity of KR significantly increased with cold-acclimation while that of PDH remained unchanged. Different electrophoretic mobilities in PAGE gel suggested that KR and PDH are two different enzymes with specific requirement for NADP(H) or NAD(H), respectively, as co-factors.

Acclimatization↗

The effects of acclimation on thermal tolerance, desiccation resistance and metabolic rate in Chirodica chalcoptera (Coleoptera: Chrysomelidae).

Despite much focus on species responses to environmental variation through space and time, many higher taxa and geographic areas remain poorly studied. We report the effects of temperature acclimation on thermal tolerance, desiccation rate and metabolic rate for adult Chirodica chalcoptera (Coleoptera: Chrysomelidae) collected from Protea nerifolia inflorescences in the Fynbos Biome in South Africa. After 7 days of acclimation at 12, 19 and 25 degrees C, critical thermal maxima (mean+/-s.e.: 41.8+/-0.2 degrees C in field-fresh beetles) showed less response (<1 degrees C change) to temperature acclimation than did the onset of the critical thermal minima (0.1+/-0.2, 1.0+/-0.2 and 2.3+/-0.2 degrees C, respectively). Freezing was lethal in C. chalcoptera (field-fresh SCP -14.6 degrees C) and these beetles also showed pre-freeze mortality. Survival of 2 h at -10.1 degrees C increased from 20% to 76% after a 2 h pre-exposure to -2 degrees C, indicating rapid cold hardening. Metabolic rate, measured at 25 degrees C and adjusted by ANCOVA for mass variation, did not differ between males and females (2.772+/-0.471 and 2.517+/-0.560 ml CO2 h(-1), respectively), but was higher in 25 degrees C-acclimated beetles relative to the field-fresh and 12 degrees C-acclimated beetles. Body water content and desiccation rate did not differ between males and females and did not respond significantly to acclimation. We place these data in the context of measured inflorescence and ambient temperatures, and predict that climate change for the region could have effects on this species, in turn possibly affecting local ecosystem functioning.

Acclimatization↗

Contributions of PIP(2)-specific-phospholipase C and free salicylic acid to heat acclimation-induced thermotolerance in pea leaves.

The relationship between the accumulation in endogenous free salicylic acid (SA) induced by heat acclimation (37 degrees C) and the activity of PIP(2)-phospholipase C (PIP(2)-PLC; EC 3.1.4.3) in the plasma membrane fraction was investigated in pea (Pisum sativum L.) leaves. We focused our attention on the hypothesis that positive SA signals induced by heat acclimation may be relayed by PIP(2)-PLC. Heat acclimation induced an abrupt elevation of free SA preceding the activation of PLC toward PIP(2). Immunoblotting indicated a molecular mass with 66.5kDa PLC plays key role in the development of thermotolerance in pea leaves. In addition, some characterizations of PLC toward PIP(2) isolated from pea leaves with two-phase purification containing calcium concentration, pH and a protein concentration were also studied. Neomycin sulfate, a well-known PIP(2)-PLC inhibitor, was employed to access the involvement of PIP(2)-PLC in the acquisition of heat acclimation induced-thermotolerance. We were able to identify a PIP(2)-PLC, which was similar to a conventional PIP(2)-PLC in higher plants, from pea leaves suggesting that PIP(2)-PLC was involved in the signal pathway that leads to the acquisition of heat acclimation induced-thermotolerance. On the basis of these results, we conclude that the involvement of free SA may function as the upstream event in the stimulation of PIP(2)-PLC in response to heat acclimation treatment.

Acclimatization↗

Tolerance and acclimation to zinc of Ceriodaphnia dubia.

Zinc is an essential metal for all living organisms. However, so far, little or no attention has been paid to the consequences of zinc deficiency or acclimation to this metal during culturing and testing on toxicity test results. In this study, the cladoceran Ceriodaphnia dubia was acclimated for 10 generations to four zinc concentrations ranging from 0 to 100 microg Zn/l and changes in zinc tolerance were monitored using acute (48 h) and chronic (9 days) assays. C. dubia deprived of zinc and acclimated to 13 microg Zn/l had a lower fitness in comparison with organisms acclimated to 50 and 100 microg Zn/l. In the two lowest versus the two highest acclimation concentrations the 9dEC50 values (on immobility) were 358-387 microg Zn/l versus 486-489 microg Zn/l; the mean number of young per female was 11-18 versus 25-32; and the time to first brood was 4.7-5.0 days versus 4.0-4.3 days. Moreover, the coefficient of variation of all parameters tested was highest in the two lowest acclimation concentrations. The results indicate that culturing test animals in media lacking trace metals such as zinc could give rise to animals that are unnaturally sensitive to those same metals daring toxicity tests.

Adaptation, Physiological↗

Photosynthetic acclimation of higher plants to growth in fluctuating light environments.

This paper describes a study into the potential of plants to acclimate to light environments that fluctuate over time periods between 15 min and 3 h. Plants of Arabidopsis thaliana (L.) Heynh., Digitalis purpurea L. and Silene dioica (L.) Clairv. were grown at an irradiance 100 mumol m(-2) s(-1). After 4-6 weeks, they were transferred to light regimes that fluctuated between 100 and either 475 or 810 mumol m(-2) s(-1), in a regular cycle, for 7 days. Plants were shown, in most cases, to be able to undergo photosynthetic acclimation under such conditions, increasing maximum photosynthetic rate. The extent of acclimation varied between species. A more detailed study with S. dioica showed that this acclimation involved changes in both Rubisco protein and cytochrome f content, with only marginal changes in pigment content and composition. Acclimation to fluctuating light, at the protein level, did not fully reflect the acclimation to continuous high light - Rubisco protein increased more than would be expected from the mean irradiance, but less than expected from the high irradiance; cytochrome f increased when neither the mean nor the high irradiance would be expected to induce an increase.

Journal Article↗

Acclimation of the diatom Stephanodiscus neoastraea and the cyanobacterium Planktothrix agardhii to simulated natural light fluctuations.

Functional and structural characteristics of the photosynthetic apparatus were studied in the diatom Stephanodiscus neoastraea and the cyanobacterium Planktothrix agardhii which were grown semi-continuously under constant irradiance or under simulated natural light fluctuations. The light fluctuations consisted of 24 oscillations of exponentially increasing and decreasing irradiance over a 12-h light period. Maximum irradiance was 1100 mumol photons m(-2) s(-1) with the ratio of maximum to minimum intensities being 100, simulating Langmuir circulations with a ratio of euphotic to mixing depth of 1. S. neoastraea acclimated to the light fluctuations by doubling the number and halving the size of photosynthetic units (PS II) while the amount of chlorophylls and carotenoids remained unchanged. The chlorophyll-specific maximum photosynthetic rate was enhanced while the slope of the photosynthesis versus irradiance curves was not influenced by the light fluctuations. Acclimation of P. agardhii was mainly characterized by an increase in chlorophyll content. Both photosystems showed only little changes in number and size. Maximum photosynthetic rate, saturating irradiance and initial slope of the photosynthesis versus irradiance curves did not vary. Although both high and low light were contained in the fluctuating light, an analogy to low or high light acclimation was not found for the diatom nor for the cyanobacterium acclimated to light fluctuations. We suggest that the acclimation to fluctuating light is a response type outside the known scheme of low and high light acclimation.

Journal Article↗

Thyroid status and noradrenaline-induced regulatory thermogenesis in heat acclimated rats.

Adjustments of the obligatory and regulatory thermogenesis upon heat acclimation (34 degrees C) and perchlorate treatment were observed in male rats. Resting oxygen consumption measured at neutral temperature was reduced, with respect to control, by 23, 27, and 34% in heat acclimated, perchlorate treated, and heat acclimated-perchlorate treated rats, respectively. Administration of 0.4 mg NA/kg increased oxygen consumption by 40, 30, 5 and 1% above resting level in control, heat acclimated, perchlorate treated, and heat acclimated-perchlorate treated rats, respectively. Whereas significant interaction was found between the effects of thyroid hormones and acclimation temperature on obligatory thermogenesis, the interaction of these factors on regulatory thermogenesis was not statistically significant. Still, hypothyroidism may be considered a thermoregulatory advantage during chronic heat exposure.

Acclimatization↗

Benefit of heat acclimation is limited by the evaporative potential when wearing chemical protective clothing.

Heat acclimation-induced sweating responses have the potential of reducing heat strain for chemical protective garment wearers. However, this potential benefit is strongly affected by the properties of the garment. If the clothing ensemble permits sufficient evaporative heat dissipation, then heat acclimation becomes helpful in reducing heat strain. On the other hand, if the garment creates an impenetrable barrier to moisture, no benefit can be gained from heat acclimation as the additional sweating cannot be evaporated. Ten subjects were studied exercising on a treadmill while wearing two different chemical protective ensembles. Skin heat flux, skin temperature, core temperature, metabolic heat production and heart rate were measured. It was found that the benefit of heat acclimation is strongly dependent on the ability of the body to dissipate an adequate amount of heat evaporatively. The evaporative potential (EP), a measure of thermal insulation modified by moisture permeability, of the clothing ensemble offers a quantitative index useful to determine, a priori, whether heat acclimation would be helpful when wearing protective clothing system. The data show that when EP is < 15%, heat acclimation affords no benefit. An evaporative potential graph is created to aid in this determination.

Acclimatization↗

Cold acclimation strategy is highly variable among the sunfishes (Centrarchidae).

We tested the hypothesis that the physiological strategy for acclimating to low body temperature is similar among closely related fish. Largemouth bass (Micropterus salmoides), green sunfish (Lepomis cyanellus), bluegill sunfish (Lepomis macrochirus), black crappie (Pomonix nigromaculatus), and white crappie (Pomonix annularis), all members of the family Centrarchidae, were acclimated to 5 degrees and 25 degrees C. Morphometric variables (total mass, total length, organ masses) and enzyme activities (hexokinase; lactate dehydrogenase; and cytochrome oxidase in heart, liver, and muscle) were measured in 5 degrees C- and 25 degrees C-acclimated fish at 5 degrees and 25 degrees C assay temperatures. Each species displayed a distinct physiological response to cold acclimation that differed among tissues. These data suggest that the response to cold acclimation is highly variable within families. Our findings are consistent with other studies suggesting that acclimation responses are labile and may evolve independently even among closely related species.

Acclimatization↗

Oxygen preference of Daphnia magna is influenced by Po2 acclimation and biotic interactions.

Oxygen preference of the planktonic crustacean Daphnia magna was observed within an oxygen gradient. Long-term acclimation to specific oxygen conditions determined the preferred oxygen tension. Animals from hypoxic acclimation stayed in oxygen-poor water at first. They chose oxygen tensions above the critical partial pressure, thus maintaining aerobic energy supply, but they avoided higher oxygen concentrations. However, they gradually took more oxygen-rich positions within 2 d. Animals from severe hypoxic and hyperoxic acclimation showed stenoxic behavior. Specimens raised in moderately hypoxic or normoxic medium spread out more widely in the oxygen gradient (euryoxic behavior). The possible role of hemoglobin and antioxidant defense capacity limiting the animals' distribution at low or high oxygen concentrations, respectively, is discussed. In tests with mixed groups of D. magna from opposing oxygen acclimation conditions, the animals clustered at intermediate oxygen conditions. However, the acclimation condition of the major group was found to determine the position taken within the gradient to a greater extent. Selected oxygen tensions were closer to the preference values of the larger group, yet the influence of the minority prevented the majority in heterogeneous groups from taking their originally determined preference position. Thus, aggregation behavior interfered with the acclimation-dependent oxygen preference behavior.

Acclimatization↗

The blue light-specific response of Vicia faba stomata acclimates to growth environment.

Stomata in epidermal strips from growth chamber-grown Vicia faba leaves opened less in response to white light than did stomata from greenhouse-grown leaves. Chlorophyll-mediated, red light-stimulated opening was similar in stomata from the two growth conditions, but stomata from the growth chamber environment had a severely reduced response to blue light. Transfer of plants between the two growth conditions resulted in an acclimation of the stomatal blue light response. Stomata lost blue light sensitivity within 1 d of transfer to growth chamber conditions and gained sensitivity to blue light over an 8 d period after transfer to a greenhouse. Short-term transfer experiments confirmed that the rapid loss of blue light sensitivity was an acclimation response, requiring between 12 and 20 h exposure to growth chamber conditions. The acclimation of the stomatal response to blue light was inversely related to a previously reported acclimation response in which stomata change between high CO2 sensitivity under growth chamber conditions and low CO2 sensitivity under greenhouse conditions. The time courses of the blue light and CO2 acclimation responses were virtually identical, suggesting the possibility of a common acclimation mechanism.

Acclimatization↗