Regulatory volume decrease in response to hypotonic stress is typical of the oocytes and early mouse embryos. Changes in the kinetics of osmotic reaction can be used as a marker of the modulating effect of the incubation medium on transmembrane transport in embryonic cells. Quantitative laser scanning microtomography (QLSM) was used to measure oocyte volume. In this paper, it is shown that addition of 5 μM glycine, taurine, or GABA, as well as ATP to Dulbecco's medium abolished the regulatory volume decrease in mature mouse oocytes.
Osmolarity of Dulbecco's medium at which the volume of two-cell mouse embryo remained similar to that of intact embryo was determined. The method is based on comparison of kinetic curves describing the volume of embryonic cell in solutions of different osmolarity. The blastomere volume was measured by quantitative laser microtomography after fixed osmotic stress intervals. It was found that Dulbecco's saline with 125 mM NaCl solution is an isotonic solution for two-cell mouse embryo. This concentration corresponds to 290 mOsm, which is lower than osmolarity (~310 mOsm) of media routinely used for culturing of differentiated cells or biological fluids, e.g. blood plasma.
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