SUMMARYThe extent to which hematocrit (Hct) is regulated and the impact of altered Hct on blood oxygen transport in avian embryos are largely unknown. Consequently, we investigated how acute blood removal or Ringer solution injection modified Hct in day·15 embryos, and how ʻblood dopingʼ with erythrocyte-enriched whole blood influenced O 2 consumption in day·15-17 chicken embryos. Mean Hct (±s.e.m.) at day·15, 16 and 17 was 26.7±0.6%, 28.0±0.4% and 30.7±0.5%, respectively. Blood withdrawal (19 increments of 125·l each, separated by 30·min) caused a progressive fall in Hct to ~12% at day·15. Hct decline was strictly proportional to the extent of blood withdrawal. Incremental Ringer solution injection over an 8·h period, transiently increasing blood volume up to 85% over initial values, did not decrease Hct, indicating that injected Ringer solution rapidly left the circulating blood compartment. Blood doping with erythrocyte-enriched whole blood artificially elevated Hct from 27% to 38%, but caused no significant change in routine O 2 consumption (0.35-0.39·ml O 2 ·min -1 ·egg -1 ) at any point over the subsequent 6·h period in day·15-17 embryos. We conclude that Hct is not protected acutely in day·15 chicken embryos, with no evidence of erythrocyte sequestration or release. Additionally, at day·15-17, Hct increases of ~10% do not enhance embryonic oxygen consumption, suggesting that blood oxygen carrying capacity per se is not limiting to oxygen consumption.Key words: hematocrit, oxygen consumption, blood volume, development, embryo.
THE JOURNAL OF EXPERIMENTAL BIOLOGY884 humidity of 60%. Eggs were turned automatically every hour. A total of 287 eggs were used in this study. Egg mass ranged from 51.07 to 77.33·g (mean ± 1 s.e.m., 58.7±0.37·g).
Venous cannulation for Hct manipulationFor withdrawal or infusion of blood or Ringer solution, a vein in the chorioallantoic membrane (CAM) was cannulated with a method adapted from that of Tazawa et al. . Briefly, each egg was candled to find the largest CAM vein at the egg's blunt end. The egg was then half-buried in a sand bath set at 38°C to maintain egg temperature throughout surgery. A piece of eggshell (~4·mm in diameter) above the selected vein was removed. The inner eggshell membrane was carefully removed to reveal the underlying vein, which was then non-occlusively cannulated in a downstream direction. The cannula comprised a 30·gauge needle, bent at 90° approximately 2·mm from the tip, which was glued into 100·mm of PE·10 tubing, which in turn was glued into 100·mm of PE·50 tubing. Prior to its insertion, the cannula was filled with heparinized (100·units·ml The open end of the cannula was then closed with a small stainless steel pin. The egg was returned to the incubator (at 38°C) immediately after cannula implantation.
Hct determinationHct was determined on 25·l of undiluted blood drawn into a Hamilton syringe through the implanted cannula. Blood volume in day·15 embryos is ~2.5·ml (see Tazawa and Whittow, 2000), so the volume of this blood sample represents ~1...