Rieger D, McGowan L T, Cox S F, Pugh P A, Thompson J G
Department of Biomedical Sciences, University of Guelph, Ontario, Canada.
Reprod Fertil Dev. 2002;14(5-6):339-43. doi: 10.1071/rd02038.
In cattle embryos, the proportion of ATP produced by glycolysis increases following the major activation of the embryonic genome, and development to the blastocyst stage is improved in the presence of 10 microM 2,4-dinitrophenol (DNP), an uncoupler of oxidative phosphorylation, from Day 5 to Day 7 of culture. In Experiment 1 of the present study, culture of cattle embryos in the presence of 10 microM DNP from Day 5 to Day 7 stimulated development to the blastocyst stage, but had no significant effects on oxygen, pyruvate or glucose uptake, or on lactate production. In Experiment 2, culture of cattle embryos in the presence of 10 microM DNP from Day 5 to Day 7, stimulated the metabolism of [2-14C]pyruvate (a measure of Krebs cycle activity) on all of Days 5, 6 and 7, and stimulated metabolism of [5-3H]glucose (a measure of glycolysis) on Day 7 only. The results show that 10 microM DNP stimulates oxidative and glycolytic metabolism in Day-5 to Day-7 cattle embryos, but this does not fully explain the observed increase in developmental competence. We propose that partial inhibition or uncoupling of oxidative phosphorylation may reduce the level of intracellular reactive oxygen species production, thereby facilitating development.
在牛胚胎中,随着胚胎基因组的主要激活,糖酵解产生的三磷酸腺苷(ATP)比例增加,并且在培养的第5天到第7天,在存在10微摩尔2,4-二硝基苯酚(DNP,一种氧化磷酸化解偶联剂)的情况下,胚胎发育到囊胚阶段的情况得到改善。在本研究的实验1中,从第5天到第7天在10微摩尔DNP存在下培养牛胚胎,刺激了其发育到囊胚阶段,但对氧气、丙酮酸或葡萄糖的摄取以及乳酸产生没有显著影响。在实验2中,从第5天到第7天在10微摩尔DNP存在下培养牛胚胎,在第5、6和7天均刺激了[2-¹⁴C]丙酮酸的代谢(三羧酸循环活性的一种度量),并且仅在第7天刺激了[5-³H]葡萄糖的代谢(糖酵解的一种度量)。结果表明,10微摩尔DNP刺激了第5天到第7天牛胚胎的氧化和糖酵解代谢,但这并不能完全解释所观察到的发育能力的提高。我们提出,氧化磷酸化的部分抑制或解偶联可能会降低细胞内活性氧的产生水平,从而促进发育。