Effects of fasting and refeeding on body mass,thermogenesis and serum leptin in Brandt's voles (Lasiopodomys brandtii)

(1)

To investigate the effect of fasting and refeeding on the body mass, thermogenesis and serum leptin in Brandt's voles, the changes in body and body fat mass, resting metabolic rate (RMR), mitochondrial cytochrome c oxidase (COX) activity in liver and brown adipose tissue (BAT), uncoupling protein 1 (UCP1) content of BAT, serum leptin level and post-fasting food intake were monitored and measured.     

Geographic variation in basal thermogenesis in little buntings: Relationship to cellular thermogenesis and thyroid hormone concentrations

Acclimatization to different ambient conditions is an essential prerequisite for survival of small passerine birds. Long-distance migration and winter acclimatization induce similar physiological and biochemical adjustments in passerines. To understand metabolic adaptations, the resting metabolic rate (RMR), the thermogenic properties of mitochondria in liver and muscle, and the activity of thyroid hormones were examined in field-captured little buntings (Emberiza pusilla) between Southeastern (Wenzhou) and Northeastern (Qiqihar) China from March to May in 2008 during their migration. Twelve birds were trapped from March to April in Wenzhou region, Zhejiang Province (27°29′N, 120°51′E) and eleven birds originated from April to May in Qiqihar region, Heilongjiang Province (47°29′N, 124°02′E). We found that RMRs of little buntings were significantly higher in Qiqihar than in Wenzhou. Consistently, mitochondrial state-4 respiration capacities and cytochrome c oxidase activities (COX) in liver and muscle, and circulating levels of plasma triiodothyronine (T₃) of little buntings were also significantly higher in Qiqihar than in Wenzhou. Variation in metabolic biochemical markers of liver and muscle, such as state-4 respiration and COX, and variation in thyroid hormone levels were correlated with variation in RMR. There was also a positive relationship between T₃ and metabolic biochemical markers. Little buntings mainly coped with a cold environment by enhancing thermogenic capacities through enhanced respiratory enzyme activities and plasma T₃. These results support the view that the primary means by which small birds meet energetic challenges of cold conditions is through metabolic adjustments.     

Suppression of thermogenic capacity during reproduction in primiparous brandt's voles (Microtus brandtii)

• (1)To investigate the changes of brown adipose tissue (BAT) thermogenic capacity in primiparous Brandt's voles during different phases of reproduction, BAT weight, mitochondrial protein concentration, cytochrome c oxidase (COX) activity, and uncoupling protein (UCP1) contents were measured.
• (2)Both cytochrome c oxidase activity and UCP1 contents decreased significantly during lactation, suggesting that thermogenic capacity was suppressed.
• (3)The decrease of thermogenic capacity during reproduction, especially during lactation, is compensation to the large demand of energy for reproduction. This is advantageous for energy conservation and lactation in Brandt's voles. UCP1 is the base of molecular thermogenesis of BAT in Brandt's voles.

     

Great adaptability of brown adipose tissue mitochondria to extreme ambient temperatures in control and cold-acclimated gerbils as compared with mice

1. The gerbil (Gerbillus campestris) is a desert rodent able to tolerate high (38 degrees C) and low (-20 degrees C) ambient temperatures, probably due to both its low resting metabolic rate in hot environment and its high peak metabolic rate in cold. 2. Measurement of mitochondrial state IV respiration and cytochrome-oxidase activity (COX) were made in interscapular brown adipose tissue (IBAT), liver and hind limb muscles of gerbils and mice of nearly equal body mass, acclimated for 4 weeks at cold ambient temperature (CA) or reared at thermoneutrality (TN). 3. The most striking difference between these two animal species appears to be in IBAT mitochondria: in TN animals, the level of state IV respiration and COX activity was lower in gerbils than in mice, but the cold acclimation-induced increase in these parameters was greater in gerbils than in mice. 4. Alternatively, in gerbils as in mice, cold acclimation induced a reduction in muscle mitochondrial COX activity. No important change due to cold acclimation was observed in liver mitochondria, either in gerbils or in mice. 5. As compared with mice, the lower state IV respiration in IBAT mitochondria from TN gerbils may explain their low RMR, whereas the higher COX activity of IBAT mitochondria from CA gerbils may explain their higher PMR. 6. As a result of this great adaptability of BAT mitochondria, the gerbil seemed to be able to live in a wide range of ambient temperatures in its natural habitat.     

Cold adaptive thermogenesis in small mammals from different geographical zones of China

The mechanisms of thermogenesis and thermoregulation were studied in the tree shrew (Tupaia belangeri) and greater vole (Eothenomys miletus) of the subtropical region, and Brandt's vole (Microtus brandti), Mongolian gerbil (Meriones unguiculatus), Daurian ground squirrel (Spermophilus dauricus) and plateau pika (Ochotona curzoniae) of the northern temperate zone. Resting metabolic rate (RMR) and non-shivering thermogenesis (NST) increased significantly in T. belangeri, E. miletus, M. brandti and M. unguiculatus after cold acclimation (4 degrees C) for 4 weeks. In T. belangeri, the increase in RMR and thermogenesis at liver cellular level were responsible for enhancing the capacity of enduring cold stress, and homeothermia was simultaneously extended. Stable body temperature in M. brandti, E. miletus, M. unguiculatus and O. curzoniae was maintained mainly through increase in NST, brown adipose tissue (BAT) mass and its mitochondrial protein content, and the upregulation of uncoupling protein (UCP1) mRNA, as well as enhancement of the activity of cytochrome C oxidase, alpha-glycerophosphate oxidase and T(4) 5'-deiodinase in BAT mitochondria. The RMR in O. curzoniae and euthermic S. dauricus was not changed, while NST significantly increased during cold exposure; the former maintained their stable body temperature and mass, while body temperature in the latter declined by 4.8 degrees C. The serum T(3) concentration or ratio of T(3)/T(4) in all the species was enhanced after cold acclimation. Results indicated that: (1) the adaptive mechanisms of T. belangeri residing in the subtropical region to cold are primarily by increasing RMR and secondly by increasing NST, and the mechanisms of thermogenesis are similar to those in tropical mammals; (2) in small mammals residing in northern regions, the adaptation to cold is chiefly to increase NST; (3) the mechanism of cold-induced thermogenesis in E. miletus residing in subtropical and high mountain regions is similar to that in the north; (4) a low RMR in warm environments and peak RMR and NST in cold environments enabled M. unguiculatus to tolerate a semi-desert climate; (5) O. curzoniae has unusually high RMR and high NST, acting mainly via increasing NST to adapt to extreme cold of the Qinghai-Tibet Plateau; (6) the adaptation of euthermic S. dauricus to cold is due to an increase in NST and a relaxed homeothermia; and lastly (7) the thyroid hormone is involved in the regulation of cold adaptive thermogenesis in all the species studied.     

Mitochondrial respiration in muscle and liver from cold-acclimated hypothyroid rats.

The effects of long-term cold exposure on muscle and liver mitochondrial oxygen consumption in hypothyroid and normal rats were examined. Thyroid ablation was performed after 8-wk acclimation to 4 degrees C. Hypothyroid and normal controls remained in the cold for an additional 8 wk. At the end of 16-wk cold exposure, all hypothyroid rats were alive and normothermic and had normal body weight. At ambient temperature (24 degrees C), thyroid ablation induced a 65% fall in muscle mitochondrial oxygen consumption, which was reversed by thyroxine but not by norepinephrine administration. After cold acclimation was reached, suppression of thyroid function reduced muscle mitochondrial respiration by 30%, but the hypothyroid values remained about threefold higher than those in hypothyroid muscle in the warm. Blockade of beta- and alpha1-adrenergic receptors in both hypothyroid and normal rats produced hypothermia in vivo and a fall in muscle, liver, and brown adipose tissue mitochondria respiration in vitro. In normal rats, cold acclimation enhanced muscle respiration by 35%, in liver 18%, and in brown adipose tissue 450% over values in the warm. The results demonstrate that thyroid hormones, in the presence of norepinephrine, are major determinants of thermogenic activity in muscle and liver of cold-acclimated rats. After thyroid ablation, cold-induced nonshivering thermogenesis replaced 3,5,3'-triiodothyronine-induced thermogenesis, and normal body temperature was maintained.     

Cold exposure does not decrease serum leptin concentration,but increases energy intake and thermogenic capacity in pregnant Brandt's voles (Lasiopodomys brandtii)

In most mammals, maternal body mass and fat mass increase during pregnancy due to hyperphagia. These physiological changes provide the fetus with energy and nutrients and prepare the mother for the high energetic demands of lactation. In the present study, metabolic changes in response to cold and pregnancy were examined in female Brandt's voles (Lasiopodomys brandtii). At 23±1 °C, the voles increased body mass and deposited body fat during pregnancy. However, at 5±1 °C pregnant voles did not deposit body fat even though energy intake increased above the level in the warm. Serum leptin concentration increased during pregnancy and was not influenced by cold exposure. Thermogenic capacity, as indicated by uncoupling protein 1 (UCP1) content in brown adipose tissue (BAT), increased in cold-exposed pregnant voles. The number and mass of fetuses were not affected by cold exposure. Our data may indicate the importance of an increased serum leptin concentration for a successful outcome of the pregnancy and also the independence of leptin secretion from body fat in pregnant voles. It also implies the need to develop central leptin resistance with respect to control of energy balance for pregnant voles.     

Physiological and biochemical thermoregulatory responses of Chinese bulbuls Pycnonotus sinensis to warm temperature: Phenotypic flexibility in a small passerine

Temperature is an important environmental cue for the regulation of thermogenesis in small birds. We tested the hypothesis that a small passerine, the Chinese bulbul Pycnonotus sinensis can decrease thermogenesis in response to warmer temperatures by decreasing body mass, the organ mass and changing physiological and biochemical processes in the liver and muscle. Captive Chinese bulbuls decreased in body mass and basal metabolic rate (BMR) during 4 weeks of acclimation to 30±1 °C. They also showed a decrease in liver, kidney and small intestine mass and reduced mitochondrial state-4 respiration and cytochrome C oxidase (COX) activity in the liver and muscle compared to 10±1 °C. Furthermore, BMR was positively correlated with body mass, organ mass (liver, kidney, small intestine and entire digestive tract), and mitochondrial state-4 respiration and cytochrome C oxidase (COX) activity in the liver and muscle. These results indicate that changes in body mass, organ mass and respiratory enzyme activity are mechanisms by which Chinese bulbuls adapt to warmer temperatures. The data support the hypothesis that such physiological flexibility is ubiquitous in birds.     

The molecular and biochemical basis of nonshivering thermogenesis in an African endemic mammal, Elephantulus myurus

Uncoupling protein 1 (UCP1) mediated nonshivering thermogenesis (NST) in brown adipose tissue (BAT) is an important avenue of thermoregulatory heat production in many mammalian species. Until recently, UCP1 was thought to occur exclusively in eutherians. In the light of the recent finding that UCP1 is already present in fish, it is of interest to investigate when UCP1 gained a thermogenic function in the vertebrate lineage. We elucidated the basis of NST in the rock elephant shrew, Elephantulus myurus (Afrotheria: Macroscelidea). We sequenced Ucp1 and detected Ucp1 mRNA and protein restricted to brown fat deposits. We found that cytochrome c oxidase activity was highest in these deposits when compared with liver and skeletal muscle. Consistent with a thermogenic function of UCP1 isolated BAT mitochondria showed increased state 4 respiration in the cold, as well as palmitate-induced, GDP-sensitive proton conductance, which was absent in liver mitochondria. On the whole animal level, evidence of thermogenic function was further corroborated by an increased metabolic response to norepinephrine (NE) injection. Cold acclimation (18 degrees C) led to an increased basal metabolic rate relative to warm acclimation (28 degrees C) in E. myurus, but there was no evidence of additional recruitment of NE-induced NST capacity in response to cold acclimation. In summary, we showed that BAT and functional UCP1 are already present in a member of the Afrotheria, but the seasonal regulation and adaptive value of NST in Afrotherians remain to be elucidated.     

Can non-shivering thermogenesis in brown adipose tissue following NA injection be quantified by changes in overlying surface temperatures using infrared thermography?

We aimed to investigate whether infra red thermography (IRT) can be used to measure and quantify non-shivering thermogenesis (NST) in the short-tailed field vole Microtus agrestis, by directly comparing it with a standard method, i.e. metabolic response following Noradrenaline injection (NA). Mean skin surface temperature overlying Brown adipose tissue (BAT) depot was 0.82 degrees C higher than mean surface temperature that did not overly BAT. The difference in temperature increased by 1.26 degrees C after NA was administered. Mean skin surface temperature overlying BAT increased by 0.32 degrees C after NA was administered; however, surface temperature decreased by 1.32 degrees C after saline was administered. Mean skin surface temperature overlying BAT did not change significantly between warm and cold acclimated voles; in contrast metabolic peak following NA injection significantly increased in cold acclimated voles. There was no significant correlation between change in surface temperature after NA injection and metabolic peak following NA injection. The results of this study suggest that IRT is not a sensitive enough method to measure changes in NST capacity in BAT following NA injection, or to detect changes in NST capacity induced by cold acclimation. However, IRT can distinguish between skin surfaces overlying BAT and skin surfaces that do not.     

Daily variation of thermoregulatory costs in laboratory mice selected for high and low basal metabolic rate

Most mammals are known to have clear circadian rhythms of body temperature (T_b) and metabolic rate. Large parts of the rhythms correspond to the oscillation of nonshivering thermogenesis (NST), dependent on visceral organ mass, and, affected by mass of brown adipose tissue (BAT). I tested whether: (1) a different levels of BMR result in respective changes of T_b values and the magnitude of daily RMR oscillations both within and below thermoneutrality; (2) the amplitude of daily variation of RMR depends on ambient temperature (T_a). I studied: (1) daily variation of body temperature at T_a of 23 °C, and (2) the rate of resting metabolism (RMR) within and below thermoneutrality at the time of minimum and increasing T_b (minimum and maximum NST capacity), in two lines of laboratory mice subjected to divergent, artificial selection toward high (HBMR) and low (LBMR) basal rate of metabolism (BMR). All mice had a clear circadian rhythm of T_b with minimum of 36.4±0.2 °C at 7:00 and maximum of 37.8±0.2 °C at 21:00. Their RMR measured below thermoneutrality exhibited significant daily variation, with the maximum between 16:00 and 19:00, when T_b was rising. Within thermoneutral zone (TNZ) I found between-line, but not between-time, differences in RMR. All between-line differences in RMR could be explained by the magnitude of BMR. I did not find any between-line differences of RMR value in temperatures below thermoneutrality. The amplitude of daily variation of RMR measured below TNZ depended neither on the T_a value nor on level of BMR (or visceral organs).     

达乌尔黄鼠产热的季节性变化

达乌尔黄鼠（Ｃｉｔｅｌｌｕｓｄａｕｒｉｃｕｓ）的产热表现出明显的季节性变化。在非冬眠期,静止代谢率（ＲＭＲ）和非颤抖性产热（ＮＳＴ）于春季最高,秋季次之,夏季最低。冬眠期,ＲＭＲ降到极低水平,只为春季的３．０％。肝脏的线粒体蛋白含量、线粒体呼吸和细胞色素Ｃ氧化酶活力在秋季显著高于其它各季。褐色脂肪组织（ＢＡＴ）的重量、线粒体蛋白含量、细胞色素Ｃ氧化酶活力和α－磷酸甘油氧化酶活力,在夏季处于一年中的最低水平,到了冬季这些指标达到一年中的最高水平。在非冬眠季节ＢＡＴ产热能力升高时,ＮＳＴ能力也相应升高,这表明ＢＡＴ产热能力的增强是ＮＳＴ能力提高的部分机制。达乌尔黄鼠血清Ｔ＿４含量在年周期中没有明显改变,冬眠时血清Ｔ＿３含量显著高于其它各季。     

Adaptive character of metabolism in Eothenomys miletus in Hengduan Mountains region during cold acclimation

Environmental factors play an important role in the seasonal adaptation of body mass and thermogenesis in small, wild mammals. The purpose of the present study was to test the hypothesis that ambient temperature was a cue to trigger the seasonal adjustments in body mass, energy intake, uncoupling protein 1 (UCP1) in brown adipose tissue (BAT), and other biochemical characteristics of Eothenomys miletus during 49 days of cold exposure. Our data demonstrated that cold acclimation induced a remarkable decrease in body mass, a significant increase in energy intake and metabolic rate, and high expression of UCP1 in BAT of E. miletus. Biochemical characteristics of BAT and liver respiration were also increased following cold acclimation. These data suggest that E. miletus reduced the body mass and increased energy intake and expenditure under cold acclimation. Increased expression of UCP1 was potentially involved in the regulation of energy metabolism and thermogenic capacity following cold acclimation.     

Thermal acclimation,mitochondrial capacities and organ metabolic profiles in a reptile (<Emphasis Type="Italic">Alligator mississippiensis</Emphasis>)

Reptiles thermoregulate behaviourally, but change their preferred temperature and the optimal temperature for performance seasonally. We evaluated whether the digestive and locomotor systems of the alligator show parallel metabolic adjustments during thermal acclimation. To this end, we allowed juvenile alligators to grow under thermal conditions typical of winter and summer, providing them with seasonally appropriate basking opportunities. Although mean body temperatures of alligators in these groups differed by approximately 10°C, their growth and final anatomic status was equivalent. While hepatic mitochondria isolated from cold-acclimated alligators had higher oxidative capacities at 30°C than those from warm-acclimated alligators, the capacities did not differ at 20°C. Cold acclimation decreased maximal oxidative capacities of muscle mitochondria. For mitochondria from both organs and acclimation groups, palmitate increased oligomycin-inhibited respiration. GDP addition reduced palmitate-uncoupled rates more in liver mitochondria from warm- than cold-acclimated alligators. In muscle mitochondria, carboxyatractyloside significantly reduced palmitate-uncoupled rates. This effect was not changed by thermal acclimation. The aerobic capacity of liver, skeletal muscle and duodenum, as estimated by activities of cytochrome c oxidase (COX), increased with cold acclimation. At acclimation temperatures, the activities of COX and citrate synthase (CS) in these organs were equivalent. By measuring COX and CS in isolated mitochondria and tissue extracts, we estimated that cold acclimation did not change the mitochondrial content in liver, but increased that of muscle. The thermal compensation of growth rates and of the aerobic capacity of the locomotor and digestive systems suggests that alligators optimised metabolic processes for the seasonally altered, preferred body temperature. The precision of this compensatory response exceeds that typically shown by aquatic ectotherms whose body temperatures are at the mercy of their habitat.     

Energy budget,oxidative stress and antioxidant in striped hamster acclimated to moderate cold and warm temperatures

The mechanism of the rate of living-free radical theory suggests that higher rate of oxidative metabolism results from greater rate of mitochondria oxidative phosphorylation, leading to a consequent increase in production of free radicals. However, the relation between metabolic rate and oxidative stress is tissue dependent in animals acclimated to cold temperatures. Here we examined oxidative stress, reflected by changes of antioxidant activity and other related markers, in striped hamsters acclimated to moderate cold (15 °C), room (23 °C) or warm temperature (30 °C) for 6 weeks, by which either higher or lower metabolic rate was induced experimentally. Energy intake and the rate of metabolism and nonshivering thermogenesis were increased at 15 °C, but decreased at 30 °C compared with that at 23 °C. Effects of temperatures on the markers of both oxidative stress and antioxidant activities were rarely significant. The percentages of positive correlation between the 11 tissues (brain, BAT, liver, heart, lung, kidneys, stomach, small and large intestine, caecum and skeletal muscle) were 14.5% (8/55) for catalase (CAT), 7.3% (4/55) for the capacity of inhibition of hydroxyl free radical (CIH), 5.5% (3/55) for activities of superoxide dismutase (SOD), 1.8% (1/55) for total antioxidant capacity (T-AOC), 4.3% (2/46) for H₂O₂ and 11.1% (4/36) for the capacity of inhibition of hydroxyl free radical (CIH). This indicated that the tissue-dependent changes of both oxidative stress and antioxidant activity were less consistent among the different tissues. Finally the data from this study were less consistent with the prediction of the mechanism of the rate of living-free radical theory.     

Homeostatic non-shivering thermogenesis in humans facts and hypotheses

This review considers current research of different forms of non-shivering thermogenesis related to thermoregulatory and substrate homeostasis. The term “homeostatic non-shivering thermogenesis (HNST)” is proposed for explanation of facultative heat production stimulated by exposure to cold, food intake and accumulation of lactate during intensive muscle loading. Similarities and differences in physiological activity are displayed in three HNST types. Existence of a number of common points makes it possible to propose common physiological mechanisms of HNST realization. Among other candidates for HNST location, the brown adipose tissue (BAT) fits best as its function is specifi between thermogenic function in cold environment and diet-induced thermogenesis that makes it possible to link these two HNST types with BAT activity. Here we present the data indirectly confirming BAT functioning in processes of homeostatic normalization not related to cold acclimation or food intake. We also consider new data about BAT functional activity, its topographic body location, mechanisms of uncoupled respiration in different tissues in adult humans and about methods of BAT diagnostics which include the use of molecular markers. We list a number of facts confirming our suggestion about BAT activity being related to homeostatic normalization after physical loading. In conclusion, we propose an experimental research program for the testing of our hypothesis regarding BAT universal homeostatic function in humans.     

Seasonal regulations of resting metabolic rate and thermogenesis in striped hamster (Cricetulus barabensis)

(1)

Resting metabolic rate (RMR), nonshivering thermogenesis (NST) and mitochondria cytochrome c oxydase (COX) activity of brown adipose tissue (BAT), as well as weight of skin and fur were measured in striped hamsters (Cricetulus barabensis) that were live-trapped in the summer, autumn, winter and spring.     

Shivering and non-shivering thermogenesis in a marsupial,the eastern barred bandicoot (Perameles gunnii)

We investigated the metabolic rate of the Tasmanian marsupial, the eastern barred bandicoot, Perameles gunnii, before and after acclimation to cold temperature (5 °C) for a 2-week period. Although body temperature did not change significantly, we observed a significant increase in the metabolic rate (MR) when measured at 5 °C before and after cold acclimation. Nor-epinephrine had a significant effect on the metabolic rate when measured in the thermoneutral zone and when measured at 5 °C after cold acclimation; however, there was no significant increase when measured at 5 °C before cold acclimation. Nor-epinephrine also resulted in a small but significant decrease in body temperature. Electromyography (EMG) measurements were obtained before and after cold acclimation during shivering. Shivering decreased after two weeks of cold exposure indicating that the bandicoot had acclimated to that temperature. Nor-epinephrine (NE) significantly reduced shivering before but not after cold acclimation. The metabolic rate and shivering decreased in the adult eastern barred bandicoot after acclimation at 5 °C and nor-epinephrine had similar effects to cold acclimation. Our findings of minor changes in thermal conductance suggest that insulation differences were unlikely explanations for our results. These experiments indicate that this marsupial is able to increase its heat production by non-shivering thermogenesis.     

Cold tolerance of UCP1-ablated mice: A skeletal muscle mitochondria switch toward lipid oxidation with marked UCP3 up-regulation not associated with increased basal,fatty acid- or ROS-induced uncoupling or enhanced GDP effects

Mice lacking the thermogenic mitochondrial membrane protein UCP1 (uncoupling protein 1) - and thus all heat production from brown adipose tissue - can still adapt to a cold environment (4 °C) if successively transferred to the cold. The mechanism behind this adaptation has not been clarified. To examine possible adaptive processes in the skeletal muscle, we isolated mitochondria from the hind limb muscles of cold-acclimated wild-type and UCP1(–/–) mice and examined their bioenergetic chracteristics. We observed a switch in metabolism, from carbohydrate towards lipid catabolism, and an increased total mitochondrial complement, with an increased total ATP production capacity. The UCP1(–/–) muscle mitochondria did not display a changed state-4 respiration rate (no uncoupling) and were less sensitive to the uncoupling effect of fatty acids than the wild-type mitochondria. The content of UCP3 was increased 3-4 fold, but despite this, endogenous superoxide could not invoke a higher proton leak, and the small inhibitory effect of GDP was unaltered, indicating that it was not mediated by UCP3. Double mutant mice (UCP1(–/–) plus superoxide dismutase 2-overexpression) were not more cold sensitive than UCP1(–/–), bringing into question an involvement of reactive oxygen species (ROS) in activation of any alternative thermogenic mechanism. We conclude that there is no evidence for an involvement of UCP3 in basal, fatty-acid- or superoxide-stimulated oxygen consumption or in GDP sensitivity. The adaptations observed did not imply any direct alternative process for nonshivering thermogenesis but the adaptations observed would be congruent with adaptation to chronically enhanced muscle activity caused by incessant shivering in these mice.     

Metabolic shifts in the Antarctic fish <Emphasis Type="Italic">Notothenia rossii</Emphasis> in response to rising temperature and <Emphasis Type="Italic">P</Emphasis> CO<Subscript>2</Subscript>

Introduction

Ongoing ocean warming and acidification increasingly affect marine ecosystems, in particular around the Antarctic Peninsula. Yet little is known about the capability of Antarctic notothenioid fish to cope with rising temperature in acidifying seawater. While the whole animal level is expected to be more sensitive towards hypercapnia and temperature, the basis of thermal tolerance is set at the cellular level, with a putative key role for mitochondria. This study therefore investigates the physiological responses of the Antarctic Notothenia rossii after long-term acclimation to increased temperatures (7°C) and elevated P CO₂ (0.2 kPa CO₂) at different levels of physiological organisation.

Results

For an integrated picture, we analysed the acclimation capacities of N. rossii by measuring routine metabolic rate (RMR), mitochondrial capacities (state III respiration) as well as intra- and extracellular acid–base status during acute thermal challenges and after long-term acclimation to changing temperature and hypercapnia. RMR was partially compensated during warm- acclimation (decreased below the rate observed after acute warming), while elevated P CO₂ had no effect on cold or warm acclimated RMR. Mitochondrial state III respiration was unaffected by temperature acclimation but depressed in cold and warm hypercapnia-acclimated fish. In both cold- and warm-exposed N. rossii, hypercapnia acclimation resulted in a shift of extracellular pH (pH_e) towards more alkaline values. A similar overcompensation was visible in muscle intracellular pH (pH_i). pH_i in liver displayed a slight acidosis after warm normo- or hypercapnia acclimation, nevertheless, long-term exposure to higher P CO₂ was compensated for by intracellular bicarbonate accumulation.

Conclusion

The partial warm compensation in whole animal metabolic rate indicates beginning limitations in tissue oxygen supply after warm-acclimation of N. rossii. Compensatory mechanisms of the reduced mitochondrial capacities under chronic hypercapnia may include a new metabolic equilibrium to meet the elevated energy demand for acid–base regulation. New set points of acid–base regulation under hypercapnia, visible at the systemic and intracellular level, indicate that N. rossii can at least in part acclimate to ocean warming and acidification. It remains open whether the reduced capacities of mitochondrial energy metabolism are adaptive or would impair population fitness over longer timescales under chronically elevated temperature and P CO₂.