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Zebrafish have become a widely used vertebrate model in physiology and reliable measures of their metabolic rate are needed. We have developed a 3D-printed respirometer and swim tunnel system and used it for obtaining accurate measurement of standard metabolic rate (SMR) and maximal, aerobic metabolic rate (MMR) in zebrafish under rest and maximal exercise, respectively. We compared a slow (stepwise) protocol to a fast (continuous) protocol for determining MMR. The fast protocol yielded slightly (but not significantly) higher oxygen consumption rates than the slow protocol and the data, in contrast to the slow protocol, followed a normal distribution. These findings point to the fast protocol as a fast and reliable method for obtaining accurate values of MMR in zebrafish. We make the 3D drawings for printing the system available to researchers, to help streamline the field of metabolic research in zebrafish and other smaller fish species.
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Consumo de Oxígeno , Impresión Tridimensional , Natación , Pez Cebra , Animales , Metabolismo Energético , Metabolismo BasalRESUMEN
Metabolic compensation has been proposed as a mean for ectotherms to cope with colder climates. For example, under the metabolic cold adaptation and the metabolic homeostasis hypotheses (MCA and MHH), it has been formulated that cold-adapted ectotherms should display both higher (MCA) and more thermally sensitive (MHH) metabolic rates (MRs) at lower temperatures. However, whether such compensation can truly be associated with distribution, and whether it interplays with cold tolerance to predict species' climatic niches, remains largely unclear despite broad ecological implications thereof. Here, we teased apart the relationship between MRs, cold tolerance and distribution, to test the MCA/MHH among 13 European ant species. We report clear metabolic compensation effects, consistent with the MCA and MHH, where MR parameters strongly correlated with latitude and climatic factors across species' distributions. The combination of both cold tolerance and MRs further upheld the best predictions of species' environmental temperatures and limits of northernmost distribution. To our knowledge, this is the first study showing that the association of metabolic data with cold tolerance supports better predictive models of species' climate and distribution in social insects than models including cold tolerance alone. These results also highlight that adaptation to higher latitudes in ants involved adjustments of both cold tolerance and MRs, to allow this extremely successful group of insects to thrive under colder climates.
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Hormigas , Hormigas/clasificación , Hormigas/fisiología , Frío , Filogenia , Metabolismo Energético , Geografía , Adaptación FisiológicaRESUMEN
In hypoxia, air-breathing fish obtain O2 from the air but continue to excrete CO2 into the water. Consequently, it is believed that some O2 obtained by air-breathing is lost at the gills in hypoxic water. Pangasionodon hypophthalmus is an air-breathing catfish with very large gills from the Mekong River basin where it is cultured in hypoxic ponds. To understand how P. hypophthalmus can maintain high growth in hypoxia with the presumed O2 loss, we quantified respiratory gas exchange in air and water. In severe hypoxia (PO2: ≈ 1.5 mmHg), it lost a mere 4.9% of its aerial O2 uptake, while maintaining aquatic CO2 excretion at 91% of the total. Further, even small elevations in water PO2 rapidly reduced this minor loss. Charting the cardiovascular bauplan across the branchial basket showed four ventral aortas leaving the bulbus arteriosus, with the first and second gill arches draining into the dorsal aorta while the third and fourth gill arches drain into the coeliacomesenteric artery supplying the gut and the highly trabeculated respiratory swim-bladder. Substantial flow changes across these two arterial systems from normoxic to hypoxic water were not found. We conclude that the proposed branchial oxygen loss in air-breathing fish is likely only a minor inefficiency.
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Bagres , Oxígeno , Animales , Dióxido de Carbono , Hipoxia/veterinaria , Transporte BiológicoRESUMEN
The giant rorqual whales are believed to have a massive food turnover driven by a high-intake lunge feeding style aptly described as the world's largest biomechanical action. This high-drag feeding behavior is thought to limit dive times and constrain rorquals to target only the densest prey patches, making them vulnerable to disturbance and habitat change. Using biologging tags to estimate energy expenditure as a function of feeding rates on 23 humpback whales, we show that lunge feeding is energetically cheap. Such inexpensive foraging means that rorquals are flexible in the quality of prey patches they exploit and therefore more resilient to environmental fluctuations and disturbance. As a consequence, the food turnover and hence the ecological role of these marine giants have likely been overestimated.
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Conducta Alimentaria , Yubarta , Animales , Fenómenos Biomecánicos , Metabolismo Energético , AlimentosRESUMEN
SIGNIFICANCE STATEMENT: During acute base excess, the renal collecting duct ß -intercalated cells ( ß -ICs) become activated to increase urine base excretion. This process is dependent on pendrin and cystic fibrosis transmembrane regulator (CFTR) expressed in the apical membrane of ß -ICs. The signal that leads to activation of this process was unknown. Plasma secretin levels increase during acute alkalosis, and the secretin receptor (SCTR) is functionally expressed in ß -ICs. We find that mice with global knockout for the SCTR lose their ability to acutely increase renal base excretion. This forces the mice to lower their ventilation to cope with this challenge. Our findings suggest that secretin is a systemic bicarbonate-regulating hormone, likely being released from the small intestine during alkalosis. BACKGROUND: The secretin receptor (SCTR) is functionally expressed in the basolateral membrane of the ß -intercalated cells of the kidney cortical collecting duct and stimulates urine alkalization by activating the ß -intercalated cells. Interestingly, the plasma secretin level increases during acute metabolic alkalosis, but its role in systemic acid-base homeostasis was unclear. We hypothesized that the SCTR system is essential for renal base excretion during acute metabolic alkalosis. METHODS: We conducted bladder catheterization experiments, metabolic cage studies, blood gas analysis, barometric respirometry, perfusion of isolated cortical collecting ducts, immunoblotting, and immunohistochemistry in SCTR wild-type and knockout (KO) mice. We also perfused isolated rat small intestines to study secretin release. RESULTS: In wild-type mice, secretin acutely increased urine pH and pendrin function in isolated perfused cortical collecting ducts. These effects were absent in KO mice, which also did not sufficiently increase renal base excretion during acute base loading. In line with these findings, KO mice developed prolonged metabolic alkalosis when exposed to acute oral or intraperitoneal base loading. Furthermore, KO mice exhibited transient but marked hypoventilation after acute base loading. In rats, increased blood alkalinity of the perfused upper small intestine increased venous secretin release. CONCLUSIONS: Our results suggest that loss of SCTR impairs the appropriate increase of renal base excretion during acute base loading and that SCTR is necessary for acute correction of metabolic alkalosis. In addition, our findings suggest that blood alkalinity increases secretin release from the small intestine and that secretin action is critical for bicarbonate homeostasis.
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Alcalosis , Bicarbonatos , Receptores Acoplados a Proteínas G , Animales , Ratones , Ratas , Alcalosis/metabolismo , Bicarbonatos/metabolismo , Ratones Noqueados , Receptores Acoplados a Proteínas G/metabolismo , Secretina , Transportadores de SulfatoRESUMEN
Temperature affects the rate of all biochemical processes in ectotherms1,2 and is therefore critical for determining their current and future distribution under global climate change3-5. Here we show that the rate of biological processes maintaining growth, homeostasis and ageing in the permissive temperature range increases by 7% per degree Celsius (median activation energy Ea = 0.48 eV from 1,351 rates across 314 species). By contrast, the processes underlying heat failure rate within the stressful temperature range are extremely temperature sensitive, such that heat failure increases by more than 100% per degree Celsius across a broad range of taxa (median Ea = 6.13 eV from 123 rates across 112 species). The extreme thermal sensitivity of heat failure rates implies that the projected increase in the frequency and intensity of heatwaves can exacerbate heat mortality for many ectothermic species with severe and disproportionate consequences. Combining the extreme thermal sensitivities with projected increases in maximum temperatures globally6, we predict that moderate warming scenarios can increase heat failure rates by 774% (terrestrial) and 180% (aquatic) by 2100. This finding suggests that we are likely to underestimate the potential impact of even a modest global warming scenario.
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Regulación de la Temperatura Corporal , Calor Extremo , Calentamiento Global , Calor , Calentamiento Global/mortalidad , Calor/efectos adversos , Envejecimiento , Crecimiento , Homeostasis , AnimalesRESUMEN
Obesity is common in captive reptiles, and reptiles are increasingly popular as companion animals and in physiological research. Obesity may present a challenge during surgical procedures using inhalation anaesthesia, as the long induction time due to the low reptilian metabolism may increase anaesthetic accumulation in the adipose tissues. This study investigated the impact of obesity on induction and recovery times from inhaled anaesthesia. The temporal change in the partial pressure of isoflurane in different tissues was predicted using a multi-compartment model. Furthermore, as right-to-left shunting can delay anaesthetic uptake and washout, we included an assessment of the combination of cardiac shunting and obesity. The model predictions indicate a clear increase in time to reach 90% equilibration of administered anaesthetic in the brain (T90) of obese non-shunting (lean 47 min, obese >100 min) and shunting (lean 81 min, obese >100 min) reptiles. The combination of obesity and shunting doubled the time to acquisition of mean anaesthetic concentration (a measure used to plan anaesthesia) from 8 min to 19 min. Adipose blood flow highly affected whether the body type had an impact on induction time, with low adipose blood flow abolishing the effect of body type. As T90 was never reached within 100 min with both the obese reptiles, it was not possible to conclude on the effect of obesity on recovery times within this study. Care should therefore be taken when anaesthetising obese reptiles for surgical purposes, to ensure adequate anaesthetic depth is attained, and recovery monitored closely.
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Anestésicos , Isoflurano , Anestesia por Inhalación , Animales , Obesidad , ReptilesRESUMEN
The lungs and kidneys are pivotal organs in the regulation of body acid-base homeostasis. In cystic fibrosis (CF), the impaired renal ability to excrete an excess amount of HCO3- into the urine leads to metabolic alkalosis [P. Berg et al., J. Am. Soc. Nephrol. 31, 1711-1727 (2020); F. Al-Ghimlas, M. E. Faughnan, E. Tullis, Open Respir. Med. J. 6, 59-62 (2012)]. This is caused by defective HCO3- secretion in the ß-intercalated cells of the collecting duct that requires both the cystic fibrosis transmembrane conductance regulator (CFTR) and pendrin for normal function [P. Berg et al., J. Am. Soc. Nephrol. 31, 1711-1727 (2020)]. We studied the ventilatory consequences of acute oral base loading in normal, pendrin knockout (KO), and CFTR KO mice. In wild-type mice, oral base loading induced a dose-dependent metabolic alkalosis, fast urinary removal of base, and a moderate base load did not perturb ventilation. In contrast, CFTR and pendrin KO mice, which are unable to rapidly excrete excess base into the urine, developed a marked and transient depression of ventilation when subjected to the same base load. Therefore, swift renal base elimination in response to an acute oral base load is a necessary physiological function to avoid ventilatory depression. The transient urinary alkalization in the postprandial state is suggested to have evolved for proactive avoidance of hypoventilation. In CF, metabolic alkalosis may contribute to the commonly reduced lung function via a suppression of ventilatory drive.
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Alcalosis/fisiopatología , Fibrosis Quística/fisiopatología , Hipoventilación/fisiopatología , Equilibrio Ácido-Base/fisiología , Alcalosis/metabolismo , Animales , Bicarbonatos/metabolismo , Antiportadores de Cloruro-Bicarbonato , Fibrosis Quística/complicaciones , Fibrosis Quística/metabolismo , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Regulador de Conductancia de Transmembrana de Fibrosis Quística/fisiología , Modelos Animales de Enfermedad , Femenino , Hipoventilación/etiología , Hipoventilación/metabolismo , Transporte Iónico , Riñón/metabolismo , Riñón/patología , Pulmón/metabolismo , Pulmón/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Eliminación Renal , Reabsorción Renal/fisiologíaRESUMEN
One of the most air-reliant obligate air-breathing fish is the South American Arapaima gigas, with substantially reduced gills impeding gas diffusion, thought to be a result of recurring aquatic hypoxia in its habitat. In normoxic water, A. gigas is reported to satisfy 70-80% of its O2 requirement from the air while excreting 60-90% of its CO2 to the water. If this pattern of gas exchange were to continue in severely hypoxic water, O2 loss at the gills would be expected. We hypothesized therefore that partitioning of CO2 would shift to the air phase in severe aquatic hypoxia, eliminating the risk of branchial O2 loss. By adapting a respirometer designed to measure aquatic MO2/MCO2, we were able to run intermittent closed respirometry on both water and air phase for both of these gasses as well as sample water for N-waste measurements (ammonia-N, urea-N) so as to calculate metabolic fuel utilization. In contrast to our prediction, we found that partitioning of CO2 excretion changed little between normoxia and severe hypoxia (83% versus 77% aquatic excretion, respectively) and at the same time there was no evidence of branchial O2 loss in hypoxia. This indicates that A. gigas can utilize distinct transfer pathways for O2 and CO2. Routine and standard MO2, N-waste excretion and metabolic fuel utilization did not change with water oxygenation. Metabolism was fuelled mostly by protein oxidation (53%), while carbohydrates and lipids accounted for 27% and 20%, respectively.
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Dióxido de Carbono , Oxígeno , Animales , Peces , Gases , Hipoxia , AguaRESUMEN
Crocodilians are unique among vertebrates in that their hemoglobin (Hb) O2 binding is allosterically regulated by bicarbonate, which forms in red blood cells upon hydration of CO2. Although known for decades, this remarkable mode of allosteric control has not yet been experimentally verified with direct evidence of bicarbonate binding to crocodilian Hb, probably because of confounding CO2-mediated effects. Here, we provide the first quantitative analysis of the separate allosteric effects of CO2 and bicarbonate on purified Hb of the spectacled caiman (Caiman crocodilus). Using thin-layer gas diffusion chamber and Tucker chamber techniques, we demonstrate that both CO2 and bicarbonate bind to Hb with high affinity and strongly decrease O2 saturation of Hb. We propose that both effectors bind to an unidentified positively charged site containing a reactive amino group in the low-O2 affinity T conformation of Hb. These results provide the first experimental evidence that bicarbonate binds directly to crocodilian Hb and promotes O2 delivery independently of CO2. Using the gas diffusion chamber, we observed similar effects in Hbs of a phylogenetically diverse set of other caiman, alligator and crocodile species, suggesting that the unique mode of allosteric regulation by CO2 and bicarbonate evolved >80-100 million years ago in the common ancestor of crocodilians. Our results show a tight and unusual linkage between O2 and CO2 transport in the blood of crocodilians, where the build-up of erytrocytic CO2 and bicarbonate ions during breath-hold diving or digestion facilitates O2 delivery, while Hb desaturation facilitates CO2 transport as protein-bound CO2 and bicarbonate.
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Caimanes y Cocodrilos , Regulación Alostérica , Animales , Bicarbonatos , Dióxido de Carbono , Hemoglobinas , OxígenoRESUMEN
Temperature tolerance is critical for defining the fundamental niche of ectotherms and researchers classically use either static (exposure to a constant temperature) or dynamic (ramping temperature) assays to assess tolerance. The use of different methods complicates comparison between studies and here we present a mathematical model (and R-scripts) to reconcile thermal tolerance measures obtained from static and dynamic assays. Our model uses input data from several static or dynamic experiments and is based on the well-supported assumption that thermal injury accumulation rate increases exponentially with temperature (known as a thermal death time curve). The model also assumes thermal stress at different temperatures to be additive and using experiments with Drosophila melanogaster, we validate these central assumptions by demonstrating that heat injury attained at different heat stress intensities and durations is additive. In a separate experiment we demonstrate that our model can accurately describe injury accumulation during fluctuating temperature stress and further we validate the model by successfully converting literature data of ectotherm heat tolerance (both static and dynamic assays) to a single, comparable metric (the temperature tolerated for 1 h). The model presented here has many promising applications for the analysis of ectotherm thermal tolerance and we also discuss potential pitfalls that should be considered and avoided using this model.
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Numerous studies have demonstrated that tardigrades in a resting state (tun state) are very resistant to exceptional stress levels in comparison with the resistance observed in multicellular organisms in general. The types of stress include desiccation and radiation, which are also relevant in astrobiological research, and therefore, tardigrades are used as multicellular model organisms. For example, tardigrades have been investigated in the TARSE, TARDIS, RoTaRad, and TARDIKISS projects; their survival has been evaluated according to stressful conditions that prevail in low earth orbit, including the effects of cosmic radiation and microgravity. Despite this interest, the study of tardigrade biology has been severely hampered by the sparsity of appropriate quantitative techniques that inform at the single-organism level. In this study, we present results on mass-specific respiration rates as a function of termination of anhydrobiosis and variations in temperature and salinity, including Mars-analog perchlorate solutions, by using microsensor technology to measure respiration. Based on our results for Richtersius cf coronifer, we estimated the activation energy (50.8 kJ/mole O2) for its metabolism as well as Q10 for selected temperature intervals. Q10 was constant-â¼1.5-between 2°C and 33°C, except for the interval 11-16°C, where Q10 was 5.5. The steady-state mass-specific respiration rate of individuals of Richtersius cf coronifer increased with increasing salinity below the lethal limit, likely representing the energy requirements of its osmoregulatory response. We report the first quantitative data of a tardigrade's metabolic dynamics during the termination of anhydrobiosis, revealing significant variation between individuals. However, we observed a general trend, that is, a high initial metabolic rate after exposure to water. Our approach would allow us to carry out quantitative physiological studies of tardigrades on board of the International Space Station, and thus significantly extend the possibility of studying the response of multicellular organisms in space. Summary statement This article reports on first measurements of mass-specific respiration rates of individual tardigrades of the species Richtersius cf coronifer during termination of anhydrobiosis as well as measurements of the impact of temperature and salinity on oxygen uptake rates.
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Salinidad , Tardigrada , Animales , Humanos , Respiración , Temperatura , AguaRESUMEN
For the past century, the importance of the Bohr effect for blood oxygen delivery has been deemed secondary to the influence of the uptake of carbon dioxide when the blood is deoxygenated (the Haldane effect). This is, however, not the case. The simultaneous oxygen and proton binding to hemoglobin can be modelled by a two-ligand, two-state formulation, while the resulting changes in acid-base status of the surrounding solution can be assessed according to Stewart's model for strong ion difference. This approach shows that an abolishment of the Bohr effect (by either equalizing pKa values of the Bohr groups of T and R states, or by removing the Bohr groups in the calculations) dramatically increases oxygen affinity, and that the Bohr effect plays a crucial role in determining the overall position and shape of the oxygen equilibrium curve. Thus, the magnitude of the Bohr effect (the Bohr factor) and oxygen affinity are directly related, and any change in hemoglobin structure that affects the Bohr factor will inevitably influence hemoglobin oxygen affinity. The modelling approach also emphasizes that pH, PCO2 and PO2 in capillaries are dependent variables, determined by arterial blood gases, the Bohr effect, the respiratory quotient (RQ) of tissue metabolism and the buffer capacity of blood. Thus, the full extent of the Bohr effect cannot be appreciated by comparing oxygen equilibrium curves made at constant PCO2 or pH, but only by comparing curves at constant proton saturation of the Bohr groups. This is because, it is the protons bound to the Bohr groups that directly influence hemoglobinoxygen binding.
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Análisis de los Gases de la Sangre/historia , Hemoglobinas/análisis , Oxígeno/sangre , Fisiología/historia , Animales , Transporte Biológico , Dióxido de Carbono/química , Hemoglobinas/química , Historia del Siglo XIX , Historia del Siglo XX , Humanos , Concentración de Iones de Hidrógeno , Ligandos , Oxígeno/química , Unión Proteica , Vertebrados/metabolismoRESUMEN
Ectotherms may experience large body temperature (Tb) variations. Higher Tb have been reported to increase baroreflex sensitivity in ectotherm tetrapods. At lower Tb, pulse interval (PI) increases and diastolic pressure decays for longer, possibly resulting in lower end-diastolic pressures and mean arterial pressures (Pm). Additionally, compensatory baroreflex-related heart rate modulation (i.e. the cardiac branch of the baroreflex response) is delayed due to increased PI. Thus, low Tb is potentially detrimental, leading to cardiovascular malfunctioning. This raises the question on how Pm is regulated in such an adverse condition. We investigated the baroreflex compensations that enables tegu lizards, Salvator merianae, to maintain blood pressure homeostasis in a wide Tb range. Lizards had their femoral artery cannulated and pressure signals recorded at 15°C, 25°C and 35°C. We used the sequence method to analyse the heart rate baroreflex-related corrections to spontaneous pressure fluctuations at each temperature. Vascular adjustments (i.e. the peripheral branch) were assessed by calculating the time constant for arterial pressure decay (τ)-resultant from the action of both vascular resistance and compliance-by fitting the diastolic pressure descent to the two-element Windkessel equation. We observed that at lower Tb, lizards increased baroreflex gain at the operating point (Gop) and τ, indicating that the diastolic pressure decays at a slower rate. Gop normalized to Pm and PI, as well as the ratio τ/PI, did not change, indicating that both baroreflex gain and rate of pressure decay are adjusted according to PI lengthening. Consequently, pressure parameters and the oscillatory power fraction (an index of wasted cardiac energy) were unaltered by Tb, indicating that both Gop and τ modulation are crucial for cardiovascular homeostasis.
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Barorreflejo/fisiología , Presión Sanguínea/fisiología , Lagartos/metabolismo , Animales , Temperatura Corporal/fisiología , Regulación de la Temperatura Corporal/fisiología , Frecuencia Cardíaca/fisiología , Lagartos/fisiología , Presorreceptores/fisiologíaRESUMEN
Studies of tardigrade biology have been severely limited by the sparsity of appropriate quantitative techniques, informative on a single-organism level. Therefore, many studies rely on motility-based survival scoring and quantifying reproductive success. Measurements of O2 respiration rates, as an integrating expression of the metabolic activity of single tardigrades, would provide a more comprehensive insight into how an individual tardigrade is responding to specific environmental factors or changes in life stages. Here, we present and validate a new method for determining the O2 respiration rate (nmol O2 mg-1 h-1) of single tardigrades under steady state, using O2 microsensors. As an example, we show that the O2 respiration rate determined in MilliQ water for individuals of Richtersius coronifer and of Macrobiotus macrocalix at 22°C was 10.8±1.84 and 13.1±2.19â nmolâ O2 mg-1 h-1, respectively.
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Tardigrada , Humanos , Oxígeno , Reproducción , AguaRESUMEN
The use of inhalational anaesthesia is ubiquitous in terrestrial vertebrates. Given the dependence of these agents on delivery by the cardiorespiratory system, we developed a new computational model predicting equilibration of inhaled anaesthetics in mammalian and ectotherm conditions including the ability of reptiles to maintain vascular shunts. A multi-compartment model was constructed from simultaneously-solved equations, verified by comparison to the literature for endo and ectotherm physiology. The time to 90% equilibration of anaesthetic in arterial blood (t90) is predicted and used to compare anaesthetics and physiologies. The five to tenfold lower cardiac output and minute ventilation of ectothermic vertebrates is predicted to slow equilibration times by five to ten times leading to 90% equilibration in ectotherm arterial blood of over 200 min, compounded by reduction in body temperature, and the extent of right-to-left vascular shunts. The impact of these findings is also influenced by the solubility coefficient of the anaesthetic, such that at net right-to-left shunt fractions of over 0.8, sevoflurane loses the advantage of faster equilibration, in comparison with isoflurane. We explore clinical strategies to regulate anaesthetic uptake in ectotherms by managing convectional flow especially by supportive ventilation and reduction of the right-to-left shunt.
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Anestésicos por Inhalación/farmacología , Corazón/efectos de los fármacos , Anestesia por Inhalación/métodos , Animales , Temperatura Corporal/efectos de los fármacos , Gasto Cardíaco/efectos de los fármacos , Isoflurano/farmacología , Mamíferos/fisiología , Respiración/efectos de los fármacos , Sevoflurano/farmacología , Ventilación/métodosRESUMEN
Zebrafish (Danio rerio) are widely used animal models. Nevertheless, the mechanisms underlying hypoxia tolerance in this species have remained poorly understood. In the present study, we have determined the effects of hypoxia on blood-O2 transport properties and mitochondrial respiration rate in permeabilized muscle fibres of adult zebrafish exposed to either 1) a gradual decrease in O2 levels until fish lost equilibrium (~1â¯h, acute hypoxia), or 2) severe hypoxia (PO2â¯â¼â¯15â¯Torr) for 48â¯h (prolonged hypoxia). Acute, short-term hypoxia caused an increase in hemoglobin (Hb) O2 affinity (decrease in P50), due to a decrease in erythrocyte ATP after erythrocyte swelling. No changes in isoHb expression patterns were observed between hypoxic and normoxic treatments. Prolonged hypoxia elicited additional reponses on O2 consumption: lactate accumulated in the blood, indicating that zebrafish relied on glycolysis for ATP production, and mitochondrial respiration of skeletal muscle was overall significantly inhibited. In addition, male zebrafish had higher hypoxia tolerance (measured as time to loss of equilibrium) than females. The present study contributes to our understanding of the adaptive mechanisms that allow zebrafish, and by inference other fish species, to cope with low O2 levels.
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Hipoxia/sangre , Músculo Esquelético/metabolismo , Consumo de Oxígeno , Oxígeno/sangre , Pez Cebra/sangre , Animales , Transporte Biológico Activo , Hemoglobinas/metabolismo , Proteínas de Pez Cebra/metabolismoRESUMEN
Air-breathing in vertebrates has evolved many times among the bony fish while in water. Its appearance has had a fundamental impact on the regulation of ventilation and acid-base status. We review the physico-chemical constraints imposed by water and air, place the extant air-breathing fish into this framework, and show how that the advantages of combining control of ventilation and acid-base status are only available to the most obligate of air-breathing fish, thus highlighting promising avenues for research.
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Espiración/fisiología , Inhalación/fisiología , Vertebrados/fisiología , Aire , Animales , Humanos , Respiración , Agua/metabolismoRESUMEN
The high blood-O2 affinity of the bar-headed goose (Anser indicus) is an integral component of the biochemical and physiological adaptations that allow this hypoxia-tolerant species to undertake migratory flights over the Himalayas. The high blood-O2 affinity of this species was originally attributed to a single amino acid substitution of the major hemoglobin (Hb) isoform, HbA, which was thought to destabilize the low-affinity T state, thereby shifting the T-R allosteric equilibrium towards the high-affinity R state. Surprisingly, this mechanistic hypothesis has never been addressed using native proteins purified from blood. Here, we report a detailed analysis of O2 equilibria and kinetics of native major HbA and minor HbD isoforms from bar-headed goose and greylag goose (Anser anser), a strictly lowland species, to identify and characterize the mechanistic basis for the adaptive change in Hb function. We find that HbA and HbD of bar-headed goose have consistently higher O2 affinities than those of the greylag goose. The corresponding Hb isoforms of the two species are equally responsive to physiological allosteric cofactors and have similar Bohr effects. Thermodynamic analyses of O2 equilibrium curves according to the two-state Monod-Wyman-Changeaux model revealed higher R-state O2 affinities in the bar-headed goose Hbs, associated with lower O2 dissociation rates, compared with the greylag goose. Conversely, the T state was not destabilized and the T-R allosteric equilibrium was unaltered in bar-headed goose Hbs. The physiological implication of these results is that increased R-state affinity allows for enhanced O2 saturation in the lungs during hypoxia, but without impairing O2 delivery to tissues.