Lopes A S, Wrenzycki C, Ramsing N B, Herrmann D, Niemann H, Løvendahl P, Greve T, Callesen H
Department of Genetics and Biotechnology, Faculty of Agricultural Sciences, University of Aahrus, DK-8830 Tjele, Denmark.
Theriogenology. 2007 Jul 15;68(2):223-36. doi: 10.1016/j.theriogenology.2007.04.055. Epub 2007 Jun 7.
Quantification of embryo respiration is a promising procedure to assess embryonic metabolism and possibly select viable embryos. At the blastocyst stage, ATP is produced by glycolysis and oxidative phosphorylation, processes that require uptake of oxygen and glucose, which is regulated by the expression of GLUT1 and G6PD. The purpose of the present study was to investigate the relationship between respiration rates and relative abundances of G6PD and GLUT1 transcripts in individual bovine blastocysts produced in vitro. Respiration rates of 104 bovine in vitro-produced blastocysts were measured individually using the nanorespirometer technology. Real-time RT-PCR was employed to determine the relative abundance of G6PD and GLUT1 mRNA in individual embryos. The mean respiration rates were similar for male and female blastocysts of the same developmental stage, but the sex ratio was skewed towards males. GLUT1 expression was down-regulated in female versus male embryos. In contrast, a approximately 1.8-fold increase in the expression of G6PD mRNA was observed in female blastocysts when compared to male blastocysts, indicating that dosage compensation for this gene had not yet occurred. Both GLUT1 and G6PD expression levels were affected by morphological quality and stage of development. Expression of GLUT1 and G6PD mRNAs was correlated with respiration rates, indicating that, in metabolically active blastocysts, uptake of oxygen and glucose are jointly increased. These findings suggest that expression of genes for oxidative phosphorylation and glycolysis are both involved in oxygen demanding ATP production.
胚胎呼吸的定量分析是评估胚胎代谢并可能筛选出有活力胚胎的一种很有前景的方法。在囊胚阶段,ATP通过糖酵解和氧化磷酸化产生,这些过程需要摄取氧气和葡萄糖,而这是由GLUT1和G6PD的表达来调节的。本研究的目的是调查体外产生的单个牛囊胚中呼吸速率与G6PD和GLUT1转录本相对丰度之间的关系。使用纳米呼吸计技术分别测量了104个体外产生的牛囊胚的呼吸速率。采用实时RT-PCR来测定单个胚胎中G6PD和GLUT1 mRNA的相对丰度。相同发育阶段的雄性和雌性囊胚的平均呼吸速率相似,但性别比例偏向雄性。与雄性胚胎相比,雌性胚胎中GLUT1的表达下调。相反,与雄性囊胚相比,雌性囊胚中G6PD mRNA的表达增加了约1.8倍,这表明该基因的剂量补偿尚未发生。GLUT1和G6PD的表达水平均受形态质量和发育阶段的影响。GLUT1和G6PD mRNA的表达与呼吸速率相关,这表明在代谢活跃的囊胚中,氧气和葡萄糖的摄取共同增加。这些发现表明,氧化磷酸化和糖酵解相关基因的表达均参与了需要氧气的ATP产生过程。