Physiological effects of hypoxia in developing and adult honeybees Apis mellifera L

The effect of hypoxia on activity of metabolism was studied in developing and adult honeybees. The feedback was established between the O2 deficit in the gas medium and its consumption by honeybees. An elevation of temperature within the limits of vital diapason for development of honeybees activated O2 consumption regardless of the hypoxia level. A prolonged action of hypoxia on the O2 physiological state was revealed. There was shown the existence of convergent similarity between the effect of O2 deficit on honeybees and on homoiothermal animals.     

Thermoregulatory and metabolic responses of Japanese quail to hypoxia

Common responses to hypoxia include decreased body temperature (T_b) and decreased energy metabolism. In this study, the effects of hypoxia and hypercapnia on T_b and metabolic oxygen consumption (V.O₂) were investigated in Japanese quail (Coturnix japonica). When exposed to hypoxia (15, 13, 11 and 9% O₂), T_b decreased only at 11% and 9% O₂ compared to normoxia; quail were better able to maintain T_b during acute hypoxia after a one-week acclimation to 10% O₂. V.O₂ also decreased during hypoxia, but at 9% O₂ this was partially offset by increased anaerobic metabolism. T_b and V.O₂ responses to 9% O₂ were exaggerated at lower ambient temperature (T_a), reflecting a decreased lower critical temperature during hypoxia. Conversely, hypoxia had little effect on T_b or V.O₂ at higher T_a (36 °C). We conclude that Japanese quail respond to hypoxia in much the same way as mammals, by reducing both T_b and V.O₂. No relationship was found between the magnitudes of decreases in T_b and V.O₂ during 9% O₂, however. Since metabolism is the source of heat generation, this suggests that Japanese quail increase thermolysis to reduce T_b. During hypercapnia (3, 6 and 9% CO₂), T_b was reduced only at 9% CO₂ while V.O₂ was unchanged.     

Metabolic and ventilatory responses to hypoxia in two altitudinal populations of the toad, Bufo bankorensis

Effects of hypoxia on resting oxygen consumption ( ), lung ventilation, and heart rate at different ambient P_O2 were compared between lowland and high altitude populations of the toad, Bufo bankorensis. Resting decreased significantly in mild hypoxia (P_O2=120 mm Hg) at 10°C and in moderate hypoxia (P_O2=80 mm Hg) at 25°C in both altitudinal populations; however, resting did not differ significantly between the two populations. Numbers of lung ventilation periods (VP) and total inspired volume (V_L) did not change with P_O2 at 10°C, but did increase at moderate and severe hypoxia (40 mm Hg), respectively, at 25°C. Resting heart rates did not change during hypoxia and did not differ between altitude populations. The results suggest (1) the effect of P_O2 change on should be considered in future studies involving transfer of anurans to a different altitude; and (2) the metabolic and ventilatory physiology in B. bankorensis does not compensate for the low temperature and P_O2 at high altitude.     

Influence of long-term hypoxia on the energy metabolism of the haemoglobin-containing bivalve Scapharca inaequivalvis: critical O2 levels for metabolic depression

Summary The oxygen consumption rate of Scapharca inaequivalvis measured under normoxic conditions over 48 h showed a significant daily cycle with lowest values occurring shortly after the dark period; all hypoxia exposure experiments were carried out during the declining part of the cycle. Animals were exposed to a constant level of hypoxia for a 12-h period in a series of 14 experiments, each at a different oxygen tension. The oxygen consumption was measured continuously, and the extent of accumulation of end-products (succinate and propionate), and the inhibitory effect of adenosine triphosphate on phosphofructokinase were determined at the end of exposures. All three parameters (oxygen consumption, end-product accumulation, phosphofructokinase inhibition) showed a remarkable correlation with major changes occurring between 2.5 and 1.5 ppm (7 and 4 kPa) O₂. The oxygen consumption rates showed a drop to 6% of the normoxic rate, but a consistent low consumption remained below 2 ppm (5.5 kPa) which partly recovered over the 12-h exposure period by about three-fold. Succinate and propionate accumulated progressively between 2.5 and 1.5 ppm (7 and 4 kPa); at [O₂]<1.5 ppm (4kPa) the concentration did not increase further, indicating that anaerobic metabolism had reached a maximum. Over the same range, phosphofructokinase showed an increased sensitivity for adenosine triphosphate, the lower inhibitor concentration at 50% V _max value pointing to depression of glycolytic rate. Despite the activation of anaerobic metabolism and the evident depression of aerobic metabolism, simple calculation demonstrates that Scapharca inaequivalvis relies mainly on aerobic metabolism even during severe hypoxia. It is assumed that the occurrence of haemoglobin in this species is essential for its capacity to survive long periods of hypoxia.Abbreviations ATP adenosine triphosphate - I₅₀ inhibitor concentration at 50% V _max - PFK phosphofructokinase - P _c critical PO₂ - SEM standard error of mean - VO₂ oxygen consumption rate - ww wet weight     

The influences of individual variations in metabolic rate and tidal conditions on the response to hypoxia in Carcinus maenas (L.)

• 1.1. The oxygen consumption of crabs in normoxic and hypoxic (50% O₂) seawater was measured directly after collection.
• 2.2. The influences of size and lunar cycles were removed by scaling the data.
• 3.3. Strong negative correlations between low individual levels of O₂ consumption and the ability to compensate for hypoxia were apparent in Wicklow (subtidal) crabs.
• 4.4. Compensation for hypoxia was much greater on the flood tide than on the ebb.
• 5.5. Crabs from Roscoff (intertidal) had lower levels of compensation than those from Wicklow.
• 6.6. Size, sex and condition had no apparent effect upon these relationships.
• 7.7. Crabs acclimated to laboratory conditions have not shown this tidal variation in compensation for hypoxia.

     

Short‐term effect of hypoxia on ammonia excretion and respiration rates in the crab carcinus maenas

The metabolic response of the crab Carcinus maenas to short‐term hypoxia (60% and 35% saturated seawater) was studied at 17.5°C in fed, 3 day‐unfed and 6 day‐unfed crabs.

Ammonia excretion rate decreased under hypoxia: a 40% and 45% decrease in the normoxic rate was observed in fed crabs at 35% saturation and in 3 day‐unfed crabs at both hypoxic levels respectively. In the 6 day‐unfed crabs, the effect of hypoxia was concealed by the effect of starvation.

Oxygen consumption rate was directly related to the external O₂ tension irrespective of the crab's nutritional state. Stressed crabs behaved as a whole, as oxygen‐conformers.

A strong relationship was observed between ammonia excretion and oxygen consumption rates in fed crabs under hypoxia but not in starved crabs.     

Effects of oxygen fraction in inspired air on force production and electromyogram activity during ergometer rowing

Six male rowers rowed maximally for 2500 m in ergometer tests during normoxia (fractional concentration of oxygen in inspired air, F _IO₂ 0.209), in hyperoxia (F _IO₂ 0.622) and in hypoxia (F _IO₂ 0.158) in a randomized single-blind fashion. Oxygen consumption (V˙O₂), force production of strokes as well as integrated electromyographs (iEMG) and mean power frequency (MPF) from seven muscles were measured in 500-m intervals. The iEMG signals from individual muscles were summed to represent overall electrical activity of these muscles (sum-iEMG). Maximal force of a stroke (F _max) decreased from the 100% pre-exercise maximal value to 67 (SD 12)%, 63 (SD 15)% and 76 (SD 13)% (P<0.05 to normoxia, ANOVA) and impulse to 78 (SD 4)%, 75 (SD 14)% and 84 (SD 7)% (P<0.05) in normoxia, hypoxia and hyperoxia, respectively. A strong correlation between F _max and V˙O₂ was found in normoxia but not in hypoxia and hyperoxia. The mean sum-iEMG tended to be lower (P<0.05) in hypoxia than in normoxia but hyperoxia had no significant effect on it. In general, F _IO₂ did not affect MPF of individual muscles. In conclusion, it was found that force output during ergometer rowing was impaired during hypoxia and improved during hyperoxia when compared with normoxia. Moreover, the changes in force output were only partly accompanied by changes in muscle electrical activity as sum-iEMG was affected by hypoxic but not by hyperoxic gas. The lack of a significant correlation between F _max and V˙O₂ during hypoxia and hyperoxia may suggest a partial uncoupling of these processes and the existence of other limiting factors in addition to V˙O₂. Accepted: 2 June 1997     

Xylem sap flow as a major pathway for oxygen supply to the sapwood of birch (Betula pubescens Ehr.)

The role of xylem sap flow as an aqueous pathway for oxygen supply to the wood parenchyma of Betula pubescens saplings was investigated. Using micro‐optode sensors the oxygen status of the sapwood was quantified in relation to mass flow of xylem sap. Sap flow was gradually reduced by an increasing oxygen depletion in the root space. The effect of sap flow on radial O₂ transport between stem and atmosphere was assessed by a stoichiometrical approach between respiratory CO₂ production and O₂ consumption. Restriction of sap flow set in 36.5 h after the onset of O₂ depletion, and was complete after 71 h. Interruption of sap flow drastically increased the O₂ deficit in the sapwood to 70%. Sap flow contributed about 60% to the total oxygen supply to the sapwood. Diurnal O₂ flow rates varied between 3 and 6.3 nmol O₂ m^?2 leaf area (LA) s^?1 during night‐ and daytime, respectively. Maximum O₂ flow rates of 20 nmol O₂ m^?2 LA s^?1 were reached at highest sap flow rates of 5.7 mmol H₂O m^?2 LA s^?1. Sap flow not only affected the oxygen status of the sapwood but also had an effect on radial O₂ transport between stem and atmosphere.     

Combined pre‐conditioning with salidroside and hypoxia improves proliferation,migration and stress tolerance of adipose‐derived stem cells

Oxidative stress after ischaemia impairs the function of transplanted stem cells. Increasing evidence has suggested that either salidroside (SAL) or hypoxia regulates growth of stem cells. However, the role of SAL in regulating function of hypoxia‐pre–conditioned stem cells remains elusive. Thus, this study aimed to determine the effect of SAL and hypoxia pre‐conditionings on the proliferation, migration and tolerance against oxidative stress in rat adipose‐derived stem cells (rASCs). rASCs treated with SAL under normoxia (20% O₂) or hypoxia (5% O₂) were analysed for the cell viability, proliferation, migration and resistance against H₂O₂‐induced oxidative stress. In addition, the activation of Akt, Erk1/2, LC3, NF‐κB and apoptosis‐associated pathways was assayed by Western blot. The results showed that SAL and hypoxia treatments synergistically enhanced the viability (fold) and proliferation of rASCs under non‐stressed conditions in association with increased autophagic flux and activation of Akt, Erk1/2 and LC3. H₂O₂‐induced oxidative stress, cytotoxicity, apoptosis, autophagic cell death and NF‐κB activation were inhibited by SAL or hypoxia, and further attenuated by the combined SAL and hypoxia pre‐treatment. The SAL and hypoxia pre‐treatment also enhanced the proliferation and migration of rASCs under oxidative stress in association with Akt and Erk1/2 activation; however, the combined pre‐treatment exhibited a more profound enhancement in the migration than proliferation. Our data suggest that SAL combined with hypoxia pre‐conditioning may enhance the therapeutic capacity of ASCs in post‐ischaemic repair.     

Generation of a hypoxia‐sensing mouse model

Oxygen (O₂) homeostasis is essential to the metazoan life. O₂‐sensing or hypoxia‐regulated molecular pathways are intimately involved in a wide range of critical cellular functions and cell survival from embryogenesis to adulthood. In this report, we have designed an innovative hypoxia sensor (O₂CreER) based on the O₂‐dependent degradation domain of the hypoxia‐inducible factor‐1α and Cre recombinase. We have further generated a hypoxia‐sensing mouse model, R26‐O₂CreER, by targeted insertion of the O₂CreER‐coding cassette in the ROSA26 locus. Using the ROSA^mTmG mouse strain as a reporter, we have found that this novel hypoxia‐sensing mouse model can specifically identify hypoxic cells under the pathological condition of hind‐limb ischemia in adult mice. This model can also label embryonic cells including vibrissal follicle cells in E13.5–E15.5 embryos. This novel mouse model offers a valuable genetic tool for the study of hypoxia and O₂ sensing in mammalian systems under both physiological and pathological conditions.     

The Effect of Mild and Severe Hypoxia on Rat Cortical Synaptosomes

Brain ischemia results in neuronal injury and neurological disability. The present study examined the effect of mild (6% O₂) and severe (2% O₂) hypoxia on mitochondria of rat cortical synaptosomes. During mild and severe hypoxia, JO₂ and ATP production significantly decreased and mitochondrial membranes depolarized. Synaptosomal calcium concentration increased slightly, albeit not significantly. After a 1 h re-oxygenation period, JO₂, ATP production and mitochondrial membrane potential returned to control levels in synaptosomes incubated in 6% O₂. In synaptosomes incubated in 2% O₂, however, the ATP production was not restored after re-oxygenation and intrasynaptosomal Ca²⁺ significantly increased. The results indicate that both mild and severe hypoxia influence the physiology of synaptosomal mitochondria; the modifications are reversible after mild hypoxia and but partly irreversible after severe hypoxia.     

Breathing, locomotion and everything in-between. Proceedings of a symposium held at the 2nd International Congress of Respiratory Science. Bad Honnef, Germany. August 9-13, 2009

In the chick embryo at day 3, gas exchange occurs by diffusion and oxygen consumption (V?_O2) does not depend on the cardiovascular convection of O₂. Whether or not this is the case in hypoxia is not known and represents the aim of the study. The heart of chicken embryos at 72 h (stage HH18) was filmed through a window of the eggshell by a camera attached to a microscope. Stroke volume was estimated from the changes in heart silhouette between systole and diastole. V?_O2was measured by a closed system methodology. In normoxia, a decrease in temperature (T) from 39 to 31 °C had parallel depressant effects on V?_O2and HR. At 39 °C, a progressive decrease in O₂ lowered V?_O2; HR was maintained until the O₂ threshold of ～ 15%. In severe hypoxia (4% O₂) V?_O2and HR were, respectively, ～ 12% and ～ 62% of normoxia. At 32 °C the hypoxic threshold for HR was significantly lower. During constant hypoxia (7% O₂) V?_O2did not respond to T, while the HR response was preserved. Stroke volume changed little with changes in T or O₂, except at 6 and 4% O₂, when it decreased by ～ 20 and 30%. In embryos growth-retarded because of incubation in chronic hypoxia, V?_O2and HR responses to T and hypoxia were similar to those of normal embryos. We conclude that in the early embryo during hypoxia cardiovascular O₂ convection is not responsible for the drop in V?_O2. The generalised hypometabolic response, in combination with the extremely small cardiac V?_O2, probably explains the minor effects of hypoxia on cardiac activity.     

Glucose uptake in mouse brain regions under hypoxic hypoxia

Glucose uptake of individual structures within the brain was studied by dry-autoradiography with 2-deoxy-D-[¹⁴C(U)]glucose under mild hypoxic hypoxia (12% O₂88% N₂ or 9% O₂91% N₂ for 1 hr). Glucose consumption in the whole brain was estimated by combined gas chromatography-mass spectrometry (GC-MS). Mild hypoxia increased the optical density of the autoradiograph in all regions. The deuterated G-6-P (Glucose-6-phosphate) synthesized from deuterated glucose decreased significantly with 9% O₂ hypoxia (P<0.05). The ratio of the deuterated G-6-P to deuterated glucose, a more appropriate indicator of glucose utilization than the concentration of deuterated G-6-P, decreased significantly with 12% O₂ hypoxia (P<0.01). The hippocampus, white matter, colliculus superior, and corpus geniculatum laterale appeared to be particulary sensitive to hypoxia.     

Root Growth and Oxygen Relations at Low Water Potentials. Impact of Oxygen Availability in Polyethylene Glycol Solutions

Polyethylene glycol (PEG), which is often used to impose low water potentials (ψ_w) in solution culture, decreases O₂ movement by increasing solution viscosity. We investigated whether this property causes O₂ deficiency that affects the elongation or metabolism of maize (Zea mays L.) primary roots. Seedlings grown in vigorously aerated PEG solutions at ambient solution O₂ partial pressure (pO₂) had decreased steady-state root elongation rates, increased root-tip alanine concentrations, and decreased root-tip proline concentrations relative to seedlings grown in PEG solutions of above-ambient pO₂ (alanine and proline accumulation are responses to hypoxia and low ψ_w, respectively). Measurements of root pO₂ were made using an O₂ microsensor to ensure that increased solution pO₂ did not increase root pO₂ above physiological levels. In oxygenated PEG solutions that gave maximal root elongation rates, root pO₂ was similar to or less than (depending on depth in the tissue) pO₂ of roots growing in vermiculite at the same ψ_w. Even without PEG, high solution pO₂ was necessary to raise root pO₂ to the levels found in vermiculite-grown roots. Vermiculite was used for comparison because it has large air spaces that allow free movement of O₂ to the root surface. The results show that supplemental oxygenation is required to avoid hypoxia in PEG solutions. Also, the data suggest that the O₂ demand of the root elongation zone may be greater at low relative to high ψ_w, compounding the effect of PEG on O₂ supply. Under O₂-sufficient conditions root elongation was substantially less sensitive to the low ψ_w imposed by PEG than that imposed by dry vermiculite.     

Thirty-five percent oxygen pre-conditioning protects PC12 cells against death induced by hypoxia

The present study is designed to investigate the effect of pre-conditioning with 35% O₂ on PC12 cell death induced by hypoxia. This study investigated whether 35% O₂ pre-conditioning for 3 h, followed by 12 h recovery, can protect PC12 cells against death induced by subsequent exposure to hypoxia for 72 h. The result showed that pre-conditioning with 35% O₂ partly blocked the decrease in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction induced by hypoxia in PC12 cells. PC12 cells pre-conditioned with 35% O₂ could generate a small quantity of reactive oxygen species (ROS), which activated the extracellular signal-regulated kinase (ERK) signalling pathway, then the over-expression of the B-cell lymphoma/leukaemia-2 (Bcl-2) was induced, which subsequently protected PC12 cell against death resulting from hypoxia exposure. In conclusion, 35% O₂ pre-conditioning could protect PC12 cells against hypoxic insult.     

Temperature and respiratory function in ectothermic vertebrates

Pulmonary ventilation is adjusted to maintain balance between O₂ demands and CO₂ elimination, which is essential for acid–base status in land ectothermic vertebrates. Rising temperatures cause increases in O₂ consumption (Q₁₀ effect) and decreases in the O₂ affinity of hemoglobin (a rightward shift in the oxygen–hemoglobin dissociation curve). These changes in air-breathing ectotherms are not proportional, i.e., the increased ventilation is relatively smaller than the change in metabolic rate. Therefore, the ratio between ventilation and metabolic rate is reduced, and consequently blood pH changes inversely with temperature. The combination of high temperatures and hypoxia exposure results in an amplified increase of ventilation, which may be explained by the balance between increased O₂-demand and decreased O₂-supply as well as increased O₂-chemoreceptors sensitivity. High temperature also increases pulmonary diffusing capacity. Global warming is expected to have significant impacts on the world’s climate, with temperature changes affecting living organisms, in relation to their physiology and distribution. These physiological mechanisms and their capacity to respond appropriately to temperature illustrate the complexity of the relationship between ambient temperature and the respiratory function in ectothermic vertebrates, which are particularly susceptible to change in their environment.     

Serotonergic sensory‐motor neurons mediate a behavioral response to hypoxia in pond snail embryos

Oxygen (O₂) is one of the most important environmental factors that affects both physiological processes and development of aerobic animals, yet little is known about the neural mechanism of O₂ sensing and adaptive responses to low O₂ (hypoxia) during development. In the pond snail, Helisoma trivolvis, the first embryonic neurons (ENC1s) to develop are a pair of serotonergic sensory‐motor cells that regulate a cilia‐driven rotational behavior. Here, we report that the ENC1‐ciliary cell circuit mediates an adaptive behavioral response to hypoxia. Exposure of egg masses to hypoxia elicited a dose‐dependent and reversible acceleration of embryonic rotation that mixed capsular fluid, thereby facilitating O₂ diffusion to the embryo. The O₂ partial pressures (Po₂) for threshold, half‐maximal, and maximal rotational response were 60, 28, and 13 mm Hg, respectively. During hypoxia, embryos relocated to the periphery of the egg masses where higher Po₂ levels occurred. Furthermore, intermittent hypoxia treatments induced a sensitization of the rotational response. In isolated ciliary cells, ciliary beating was unaffected by hypoxia, suggesting that in the embryo, O₂ sensing occurs upstream of the motile cilia. The rotational response of embryos to hypoxia was attenuated by application of the serotonin receptor antagonist, mianserin, correlated to the development of ENC1‐ciliary cell circuit, and abolished by laser‐ablation of ENC1s. Together, these data suggest that ENC1s are unique oxygen sensors that may provide a good single cell model for the examination of mechanistic, developmental, and evolutionary aspects of O₂ sensing. © 2002 Wiley Periodicals, Inc. J Neurobiol 52: 73–83, 2002     

Characterizing the escape response of juvenile summer flounder Paralichthys dentatus to diel‐cycling hypoxia

Swimming speed, angular correlation and expected displacement were measured in juvenile summer flounder Paralichthys dentatus acclimated to either oxygen saturation (c. 7·8 mg O₂ l^?1; saturation‐acclimated fish) or diel‐cycling hypoxia (cycling between 11·0 and 2·0 mg O₂ l^?1) for 10 days and subsequently exposed to more severe diel‐cycling hypoxia (cycling between 7·0 and 0·4 mg O₂ l^?1). Saturation‐acclimated P. dentatus exhibited an active response to declining dissolved oxygen (DO) by increasing swimming speed, angular correlation and expected displacement to peak levels at 1·4 mg O₂ l^?1 that were 3·5, 5·5 and 4·2 fold, respectively, greater than those at DO saturation. Diel‐cycling hypoxia‐acclimated P. dentatus also exhibited an active response to declining DO, although it was relatively less pronounced. Diel‐cycling hypoxia‐acclimated P. dentatus swimming speed, however, still doubled as DO decreased from 7·0 to 2·8 mg O₂ l^?1. Diel‐cycling hypoxia‐acclimated P. dentatus did not recover as well from low DO exposure as did saturation‐acclimated fish. This was reflected in their relatively more random swimming (low angular correlation between successive moves) and poor maintenance of rank order between individuals during the recovery phase. Even saturation‐acclimated P. dentatus did not resume swimming at speeds observed at saturation until DO was 4·2 mg O₂ l^?1. Paralichthys dentatus were very sensitive to decreasing DO, even at DO levels that were not lethal or growth limiting. This sensitivity and their poor recovery may preclude juvenile P. dentatus from using highly productive nursery habitats affected by diel‐cycling hypoxia.     

Respiratory and cardiovascular responses to acute hypoxia and hyperoxia in internally pipped chicken embryos

During the first day of hatching, the developing chicken embryo internally pips the air cell and relies on both the lungs and chorioallantoic membrane (CAM) for gas exchange. Our objective in this study was to examine respiratory and cardiovascular responses to acute changes in oxygen at the air cell or the rest of the egg during internal pipping. We measured lung (V·_O2lung) and CAM (V·_O2CAM) oxygen consumption independently before and after 60 min exposure to combinations of hypoxia, hyperoxia, and normoxia to the air cell and the remaining egg. Significant changes in V·_O2total were only observed with combined egg and air cell hypoxia (decreased V·_O2total) or egg hyperoxia and air cell hypoxia (increased V·_O2total). In response to the different O₂ treatments, a change in V·_O2lung was compensated by an inverse change in V·_O2CAM of similar magnitude. To test for the underlying mechanism, we focused on ventilation and cardiovascular responses during hypoxic and hyperoxic air cell exposure. Ventilation frequency and minute ventilation (V_E) were unaffected by changes in air cell O₂, but tidal volume (V_T) increased during hypoxia. Both V_T and V_E decreased significantly in response to decreased P_CO2. The right-to-left shunt of blood away from the lungs increased significantly during hypoxic air cell exposure and decreased significantly during hyperoxic exposure. These results demonstrate the internally pipped embryo's ability to control the site of gas exchange by means of altering blood flow between the lungs and CAM.