Non-invasive imaging of blood cell concentration and blood distribution in zebrafish Danio rerio incubated in hypoxic conditions in vivo.

This is the first study to use a combination of digital imaging techniques and vital video microscopy to study hypoxia-induced changes in blood cell concentration, angiogenesis and blood redistribution in entire animals. Zebrafish Danio rerio, which are known to be independent of convective oxygen transport until about 2 weeks post-fertilization, were raised under chronic hypoxia (P(O(2))=8.7 kPa) starting at 1 day after fertilization (d.p.f.) until 15 d.p.f. In control animals, the concentration of red cells (i.e. the number of red cells per nl blood) remained constant until 7 d.p.f., and than decreased by approximately 70% until 15 d.p.f. In hypoxic animals, however, the concentration of red cells remained significantly elevated compared to control animals at 12 and 15 d.p.f. Assuming that the hemoglobin content of the red cells is similar, hypoxic animals have a higher oxygen carrying capacity in their blood. Red cell distribution within the various parts of the circulatory system, taken as an indicator for blood distribution, revealed a significant modification in the number of blood cells perfusing the organs in hypoxic animals. At 12 d.p.f., gut perfusion was reduced by almost 50% in hypoxic animals, while perfusion of the segmental muscle tissue was increased to 350% of control values. No significant changes in brain perfusion were observed under these conditions. At 15 d.p.f., the reduction in gut perfusion was abolished, although muscle perfusion was still significantly elevated. At this time, growth of hypoxic animals was less compared to control animals, revealing that hypoxia had become deleterious for further development. The vascular bed of various organs was not obviously different in hypoxic animals compared to normoxic animals.