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The mechanisms, dynamics and effects of hypoxia acclimation were studied in the water flea Daphnia magna over three successive generations (parental, first and second filial generation: P, F1 and F2). The P generation was raised under normoxic conditions at 20 degrees C and became exposed to environmental hypoxia (10-19% air saturation) at maturity. Their progenies (F1 and F2) experienced hypoxia from birth onwards. Controls were kept under normoxic conditions. Individuals were successively sampled in a 3-day interval from each acclimation group to determine morpho-physiological parameters relevant in oxygen transport and regulation. Hypoxia acclimation induced adjustments at the haemoglobin (Hb) and metabolic level (within 3 days) but none at the systemic level. The convective performance and oxygen-sensitive control of the ventilatory and circulatory systems were the same in both acclimation groups. The Hb concentration and oxygen affinity increased by 266% and 32%, respectively. The 22% decrease in mass-specific oxygen consumption rate reduced the energy allocation to somatic growth without greatly affecting reproduction. The onset and duration of hypoxic exposure during ontogenesis have had a significant influence on Hb oxygen affinity and body size. Transgenerational effects of hypoxia acclimation could not be observed. The adjustments at the Hb and metabolic levels in combination with the smaller body size, which is advantageous to diffusive oxygen transport, reduced the critical ambient oxygen tension by approximately 50%.

The quantity and quality of the haemoglobin (Hb) of Daphnia magna is related to oxygen partial pressure in the water. Both the dynamics of hypoxia-induced Hb gene transcription, as well as Hb properties in animals incubated long-term at hyperoxia, normoxia and hypoxia, were investigated. Examination of Hb gene (dhb1-dhb3) transcription showed the expression of dhb2 and especially dhb3 to increase markedly approximately one hour after the onset of hypoxia, whereas dhb1 was expressed more or less constitutively. At an incubation close to anoxia, an onset of dhb3 transcription was found already after two minutes. In long-term incubated animals, concentration and oxygen affinity of Hb were lower at higher oxygen partial pressures. With decreasing oxygen availability, the subunit composition of Hb macromolecules changed. The share of the dhb2-encoded subunit, DHbF, increased already during moderate hypoxia. The increase of dhb3 mRNA (encoding DHbC) may be related to a transient increase of DHbC in the first days of hypoxia and/or to an additional coding of dhb3 for DHbD. The rise of DHbD, and particularly DHbA, only at severe hypoxia coincided with the increase of Hb oxygen affinity. The dhb1-encoded subunits DHbB and DHbE showed either a relatively moderate increase or even a decrease in concentration at hypoxia. In small animals with restricted homeostasis capabilities such as Daphnia, adaptation of the protein equipment seems to be a more effective strategy than allosteric modulator control.

To evaluate the role of temperature in oxygen transport in the water flea Daphnia magna, temperaturerelated behaviour as well as oxygen-transport physiology and biochemistry were investigated in animals long-term acclimated to moderate temperatures: 10, 20, or 30°C (normoxia) or 20°C (hypoxia). Study of the behaviour of animal groups within a normoxic thermal gradient showed their preferred temperatures and distribution patterns to be connected with acclimation conditions; for example, cold-acclimated individuals, with their relatively high metabolic rates and low oxygen-transport capacities, crowded at low temperatures where the oxygen concentration was high. One key to explaining these data is to assume a strategy of avoiding oxygen shortage. Both temperature and hypoxia acclimation also modified oxygen-transport variables such as oxygen consumption, ventilation and circulation rates, or the concentration and oxygen affinity of haemoglobin. Characteristic features of cold-acclimated D. magna were relatively high oxygen-consumption and heart rates and a low concentration and oxygen affinity of haemoglobin. Hypoxia-acclimated D. magna showed relatively low convective rates (frequency of thoracic-limb movements, heart rate) but a high concentration and oxygen affinity of haemoglobin. Additional experiments showed that independently of each other, temperature and ambient oxygen concentration modify haemoglobin quantity and quality, the former possibly acting via a temperature-induced hypoxia. The varying oxygen affinity is probably due to alterations of haemoglobin subunit composition, with the particular participation of subunits A, E, and G as revealed by two-dimensional gel electrophoresis.Résumé : L'étude du comportement relié à la température ainsi que de la physiologie et de la biochimie du transport de l'oxygène chez des cladocères Daphnia magna acclimatés à long terme à des températures modérées -10, 20, 30°C (en normoxie) ou 20 EC (en anoxie) -nous a permis d'évaluer le rôle de la température en rapport avec le transport de l'oxygène. Le comportement de groupes d'animaux dans un gradient thermique normoxique montre que les choix de température et les patterns de répartition sont reliés aux conditions d'acclimatation; par exemple, les individus acclimatés au froid, qui ont des taux métaboliques relativement élevés et des capacités réduites de transport d'oxygène, se regroupent dans les zones de température préférentielle basse où les concentrations d'oxygène sont éle-vées. Une façon d'expliquer ces données est d'assumer qu'il y a une stratégie d'évitement des pénuries d'oxygène. L'acclimatation tant à la température qu'à l'hypoxie modifie aussi les variables reliées au transport de l'oxygène, telles que la consommation d'oxygène, les taux de ventilation et de circulation ou alors la concentration de l'hémoglobine et son affinité pour l'oxygène. Les caractéristiques des cladocères acclimatés au froid comprennent une consommation d'oxygène et un rythme cardiaque relativement élevés et une concentration basse ...