Butcher L, Coates A, Martin K L, Rutherford A J, Leese H J
Department of Biology, University of York, United Kingdom.
Biol Reprod. 1998 Apr;58(4):1054-6. doi: 10.1095/biolreprod58.4.1054.
Pyruvate is added to all media used for human in vitro fertilization and embryo culture, but its function(s) in the early embryo is unknown. We tested the possibility that pyruvate can act as an oxidizable energy source by measuring the consumption of pyruvate and oxygen by Day 2 and Day 3 human embryos, using microfluorometric techniques. Oxygen consumption (19.6 pmol/embryo per hour) could account for the oxidation of only 56% of the pyruvate consumed (13.9 pmol/embryo per hour). Oxygen was also consumed in the absence of exogenous substrates. Lactate appeared in the incubation medium with pyruvate (0.47 mM) as sole exogenous substrate at a rate of 12.1 pmol/embryo per hour, at a similar rate (10.85 pmol/embryo per hour) in the presence of 1 mM glucose and 0.47 mM pyruvate, and at 2.25 pmol/embryo per hour in the absence of exogenous substrates, suggesting that a high proportion of the pyruvate taken up by early human embryos is converted to lactate. Pyruvate uptake in the presence of UK5099, a pyruvate transport inhibitor, was reduced to 10% of control values, consistent with the presence of the monocarboxylate carrier in the human embryo plasma membrane.
丙酮酸被添加到用于人类体外受精和胚胎培养的所有培养基中,但其在早期胚胎中的功能尚不清楚。我们使用微荧光技术,通过测量第2天和第3天人类胚胎对丙酮酸和氧气的消耗,来测试丙酮酸是否可以作为一种可氧化的能量来源。氧气消耗(每小时19.6皮摩尔/胚胎)仅占所消耗丙酮酸(每小时13.9皮摩尔/胚胎)氧化量的56%。在没有外源底物的情况下也会消耗氧气。当丙酮酸(0.47 mM)作为唯一外源底物时,乳酸以每小时12.1皮摩尔/胚胎的速率出现在孵育培养基中;在存在1 mM葡萄糖和0.47 mM丙酮酸的情况下,乳酸出现的速率相似(每小时10.85皮摩尔/胚胎);在没有外源底物的情况下,乳酸出现的速率为每小时2.25皮摩尔/胚胎,这表明早期人类胚胎摄取的丙酮酸中有很大一部分转化为了乳酸。在丙酮酸转运抑制剂UK5099存在的情况下,丙酮酸摄取量降至对照值的10%,这与人类胚胎质膜中存在单羧酸载体一致。