Johnson M D, Batey D W, Behr B, Barro J
Department of Gynecology and Obstetrics, Stanford University School of Medicine, CA 94305-5317, USA.
Mol Hum Reprod. 1997 Apr;3(4):351-7. doi: 10.1093/molehr/3.4.351.
In mouse and human preimplantation development, pyruvate is consumed preferentially during early embryogenesis; however, during the morula and blastocyst stages, glucose is the preferred energy substrate. Studies have suggested that the glycolytic enzymes, hexokinase and glucose phosphate isomerase, are important enzymes in glucose metabolism during these later stages of human and mouse preimplantation development. In order to investigate the genetic activities of these enzymes in late-stage mouse embryos developing in vitro, we analysed hexokinase and glucose phosphate isomerase transcription activities by qualitative RNA assays using reverse transcriptase-nested polymerase chain reaction amplification of individual mouse morulae and early blastocysts incubated in glucose/phosphate-free preimplantation stage one (P1) medium and glucose/phosphate-containing human tubal fluid (HTF) medium. We observed an increased incidence of hexokinase transcripts in the population of blastocysts compared with morulae, and differences in transcript incidence between early blastocysts developing in HTF medium and in P1 medium. In contrast, glucose phosphate isomerase transcripts were consistantly present in all embryos analysed, and appear to be constitutively expressed during late-stage mouse embryogenesis. The different activity patterns of the two glycolytic genes may reflect different mechanisms of gene regulation or differential transcript stability during the later stages of mouse preimplantation development.
在小鼠和人类的植入前发育过程中,丙酮酸在早期胚胎发生期间被优先消耗;然而,在桑葚胚和囊胚阶段,葡萄糖是首选的能量底物。研究表明,糖酵解酶己糖激酶和葡萄糖磷酸异构酶是人类和小鼠植入前发育后期葡萄糖代谢中的重要酶。为了研究这些酶在体外发育的晚期小鼠胚胎中的基因活性,我们通过定性RNA分析,使用逆转录-巢式聚合酶链反应扩增单个小鼠桑葚胚和早期囊胚,在不含葡萄糖/磷酸盐的植入前阶段一(P1)培养基和含葡萄糖/磷酸盐的人输卵管液(HTF)培养基中培养,分析了己糖激酶和葡萄糖磷酸异构酶的转录活性。我们观察到,与桑葚胚相比,囊胚群体中己糖激酶转录本的发生率增加,并且在HTF培养基和P1培养基中发育的早期囊胚之间转录本发生率存在差异。相比之下,葡萄糖磷酸异构酶转录本在所有分析的胚胎中持续存在,并且在晚期小鼠胚胎发生过程中似乎是组成性表达的。这两个糖酵解基因的不同活性模式可能反映了小鼠植入前发育后期基因调控的不同机制或转录本稳定性的差异。