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早期小鼠胚胎的氧消耗与能量代谢

Oxygen consumption and energy metabolism of the early mouse embryo.

作者信息

Houghton F D, Thompson J G, Kennedy C J, Leese H J

机构信息

Department of Biology, University of York, UK.

出版信息

Mol Reprod Dev. 1996 Aug;44(4):476-85. doi: 10.1002/(SICI)1098-2795(199608)44:4<476::AID-MRD7>3.0.CO;2-I.

Abstract

Oxygen consumption of preimplantation and early postimplantation mouse embryos has been measured using a novel noninvasive ultramicrofluorescence technique, based on an oil-soluble, nontoxic quaternary benzoid compound pyrene, whose fluorescence is quenched in the presence of oxygen. Pyruvate and glucose consumption, lactate production, and glycogen formation from glucose were also measured. Preimplantation mouse embryos of the strain CBA/Ca x C57BL/6 were cultured in groups of 10-30 in 2 microliters of modified M2 medium containing 1 mmol l-1 glucose, 0 mmol l-1 lactate, and 0.33 mmol l-1 pyruvate, for between 4-6 hr. Day 6.5 and 7.5 embryos were cultured singly in 40 microliters M2 medium for between 2-3 hr. Oxygen consumption was detected at all stages of development, including, for the first time, in the early postimplantation embryo. Consumption remained relatively constant from zygote to morula stages before increasing in the blastocyst and day 6.5-7.5 stages. When expressed as QO2 (microliters/mg dry weight/hr), oxygen consumption was relatively constant from the one-cell to morula stages before increasing sharply at the blastocyst stage and declining to preblastocyst levels on days 6.5 and 7.5. Pyruvate was consumed during preimplantation stages, with glucose uptake undetectable until the blastocyst stage. Glucose was the main substrate consumed by the 6.5 and 7.5 day embryo. The proportions of glucose accounted for by lactate appearance were 81%, 86%, and 119% at blastocyst, day 6.5, and day 7.5 stages, respectively. The equivalent figures for glucose incorporated into glycogen were 10.36%, 0.21%, and 0.19%, respectively. The data are consistent with a switch from a metabolism dependent on aerobic respiration during early preimplantation stages to one dependent on both oxidative phosphorylation and aerobic glycolysis at the blastocyst stage, a pattern which is maintained on days 6.5 and 7.5. Our technique for measuring oxygen consumption may have diagnostic potential for selecting viable embryos for transfer following assisted conception techniques in man and domestic animals.

摘要

利用一种新型非侵入性超微荧光技术,对植入前和植入后早期小鼠胚胎的耗氧量进行了测量。该技术基于一种油溶性、无毒的季铵苯化合物芘,其荧光在有氧存在时会被淬灭。同时还测量了丙酮酸和葡萄糖的消耗量、乳酸的产生量以及葡萄糖形成糖原的情况。将CBA/Ca×C57BL/6品系的植入前小鼠胚胎以10 - 30个为一组,培养在2微升含有1毫摩尔/升葡萄糖、0毫摩尔/升乳酸和0.33毫摩尔/升丙酮酸的改良M2培养基中,培养4 - 6小时。第6.5天和第7.5天的胚胎单独培养在40微升M2培养基中,培养2 - 3小时。在发育的各个阶段都检测到了耗氧量,首次包括植入后早期胚胎。从合子到桑椹胚阶段,耗氧量相对恒定,然后在囊胚期和第6.5 - 7.5天阶段增加。当以QO2(微升/毫克干重/小时)表示时,从单细胞到桑椹胚阶段耗氧量相对恒定,在囊胚期急剧增加,在第6.5天和第7.5天降至囊胚前期水平。在植入前阶段消耗丙酮酸,直到囊胚期才检测到葡萄糖摄取。葡萄糖是第6.天和第7.5天胚胎消耗的主要底物。在囊胚期、第6.5天和第7.5天阶段,葡萄糖转化为乳酸的比例分别为81%、86%和119%。葡萄糖掺入糖原的相应数字分别为10.36%、0.21%和0.19%。这些数据与从植入前早期依赖有氧呼吸的代谢转变为囊胚期依赖氧化磷酸化和有氧糖酵解的代谢一致,这种模式在第6.5天和第7.5天得以维持。我们测量耗氧量的技术可能具有诊断潜力,可用于在人类和家畜的辅助受孕技术后选择可存活的胚胎进行移植。

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