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体外受精和孤雌激活后山羊植入前胚泡的葡萄糖和丙酮酸代谢

Glucose and pyruvate metabolism of preimplantation goat blastocysts following in vitro fertilization and parthenogenetic activation.

作者信息

Ongeri E M, Krisher R L

机构信息

Department of Animal Sciences, Purdue University, West Lafayette, Indiana 47907-1151, USA.

出版信息

Cloning Stem Cells. 2001;3(3):115-23. doi: 10.1089/153623001753205070.

Abstract

The energy metabolism of preimplantation embryos can be used to predict viability and postimplantation development. Although preimplantation development and mean blastocyst cell numbers of goat in vitro-fertilized (IVF) embryos and chemically activated parthenogenotes are comparable, mammalian parthenogenotes are not viable, with most dying shortly after implantation. The objective of this study was to compare glucose and pyruvate metabolism of IVF goat blastocysts with that of parthenogenetic blastocysts developing from chemically activated oocytes. Embryos derived from IVF and parthenogenotes produced by exposing oocytes to either ionomycin or ethanol followed by 6-dimethylaminopurine (6-DMAP) were cultured in G1.2/G2.2 sequential culture media. Metabolism was determined for individual blastocysts using [5-3H]glucose and [2-14C]pyruvate to determine glycolytic and Kreb's cycle activity, respectively. Data were analyzed by ANOVA. A significantly higher percentage of activated oocytes underwent cleavage and developed to the blastocyst stage compared to IVF oocytes (p < 0.05). There was no significant difference in glucose or pyruvate metabolism between IVF and parthenogenetically activated blastocysts. Mean glucose metabolism through glycolysis was 154.9 +/- 29.1, 130.3 +/- 17.1, and 129 +/- 16.5 pmol/embryo/3 h for IVF, ethanol-activated, and ionomycin-activated blastocysts, respectively. Mean pyruvate metabolism through the Kreb's cycle was 28.1 +/- 8.0, 15.8 +/- 4.2, and 24.4 +/- 4.4 in pmol/embryo/3 h for IVF, ethanol-activated, and ionomycin-activated blastocysts, respectively. Our results suggest that known differences in postimplantation development observed in IVF versus parthenogenetic embryos cannot be attributed to differences in pyruvate or glucose metabolism in the preimplantation blastocysts. Thus, these activation protocols result in embryos capable of appropriate regulation of key metabolic enzymes.

摘要

植入前胚胎的能量代谢可用于预测其活力和植入后发育情况。尽管山羊体外受精(IVF)胚胎和化学激活孤雌生殖胚胎的植入前发育及平均囊胚细胞数量相当,但哺乳动物孤雌生殖胚胎无法存活,大多数在植入后不久死亡。本研究的目的是比较IVF山羊囊胚与化学激活卵母细胞发育而来的孤雌生殖囊胚的葡萄糖和丙酮酸代谢。将IVF获得的胚胎以及通过用离子霉素或乙醇处理卵母细胞后再用6-二甲基氨基嘌呤(6-DMAP)处理产生的孤雌生殖胚胎在G1.2/G2.2序贯培养基中培养。使用[5-³H]葡萄糖和[2-¹⁴C]丙酮酸分别测定单个囊胚的糖酵解和三羧酸循环活性,以确定代谢情况。数据采用方差分析。与IVF卵母细胞相比,激活的卵母细胞发生卵裂并发育到囊胚阶段的比例显著更高(p < 0.05)。IVF囊胚和孤雌激活囊胚之间的葡萄糖或丙酮酸代谢没有显著差异。通过糖酵解的平均葡萄糖代谢量分别为:IVF囊胚154.9±29.1、乙醇激活囊胚130.3±17.1、离子霉素激活囊胚129±有16.5 pmol/胚胎/3小时。通过三羧酸循环的平均丙酮酸代谢量分别为:IVF囊胚28.1±8.0、乙醇激活囊胚15.8±4.2、离子霉素激活囊胚24.4±4.4 pmol/胚胎/3小时。我们的结果表明,IVF胚胎与孤雌生殖胚胎在植入后发育中观察到的已知差异不能归因于植入前囊胚中丙酮酸或葡萄糖代谢的差异。因此,这些激活方案产生的胚胎能够对关键代谢酶进行适当调节。

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