Hexokinase Activity of Bovine Embryos Produced from IVM-IVF System and Their Subsequent Development

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To investigate the Embden-Meyarhof pathway (EMP) in bovine embryos, individual 1-cell to.blastocyst stage embyos produced from in vitro maturation (IVM) and in vitro fertilization (IVF) systems were subjected tomeasure hexokinase activity using a cyclinf enzyme method. In the present study, we founded that Ca++, Mg++-free phosphate buffered saline, PBS (-), coild be used as a reaction solution instead of Tris - HCI buffer and that after measurement the viability of the embryo could be maintained follwed by subsequent development. When measured by adding glucose as a substrate, hexokinase activity was 227. 9±18.6, 261.9±19.0, 337.8±29.8, 241.8±37.7 and 252.2±17.7x10-9 mol/embryo/hr in the 2-cell, 4-cell, 8-cell, morula and blastocyst stage embryos, raspectively, indicating a significant difference between embryos at the 2-cell stage and those at more abvanced stages (P<0.05). In embryos at all stages examined, the level of hexokinase activity was increased by adding the substrate. When subsequent development after measurement was examined using the 4-cell stage embryos, the development rate to the blastocyst stage was not significantly correlated to the hexokinase activity. The results suggest that the onset of glucose metabolism in the early embryos may be directly related to the synthesis of one or more key glycolytic enzyme.
日本哺乳動物卵子学会の論文