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The formation of a functional cardiovascular system is an essential step in the early vertebrate embryo. Nevertheless, the effect of hypoxia on the developmental program of organisms was studied rarely. In particular, this holds true for vertebrate embryos that depend on a functional placenta for proper development and had not been studied in this respect due to the obvious limitation. We established a protocol to culture aquatic embryos, which enabled us to culture a high number of Xenopus embryos until tadpole stage under defined hypoxic conditions in four hypoxia chambers simultaneously, employing a computerized system. In general, our results show that hypoxia results in delayed development and, in particular, we could show that oxygen availability was most crucial during gastrulation and organogenesis (early tailbud) phases during embryonic development of Xenopus laevis.
Figure 1. Experimental setup.Notes: (A) Schematic representation of hypoxia chamber setup. 1, plastic chamber; 2, 0.1× MBS buffer; 3, 10 cm Petri dish; 4, embryos; 5, glass beaker; 6, oxygen probe; 7, air-stone; 8, stirrer; 9, nitrogen gas supply; 10, OXYREG device; 11, computer. (B) Running hypoxia experiment. Four hypoxia chambers were regulated at 4% oxygen along with a normoxia chamber. Temperatures in the chambers were maintained with the help of thermostat placed below the chambers. Regulation was computer controlled. (C) Growing embryos in hypoxia starting at various developmentally sensitive stages. Embryos were placed in hypoxia chambers at gastrulation, neurulation, and tailbud stages and harvested at a later point during development for assessment.Abbreviation: MBS, Modified Barth's Solution.
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