Mognetti B, Leppens G, Sakkas D
Laboratoire des Gamètes, Clinique de Stérilité, Hôpital Cantonal Universitaire de Genève, Switzerland.
Mol Reprod Dev. 1996 Apr;43(4):421-7. doi: 10.1002/(SICI)1098-2795(199604)43:4<421::AID-MRD3>3.0.CO;2-R.
Mouse preimplantation embryo development is characterized by a switch from a dependence on the tricarboxylic acid cycle pre-compaction to a metabolism based on glycolysis post-compaction. In-view of this, the role of glucose in embryo culture medium has come under increased analysis and has lead to improved development of outbred mouse embryos in glucose free medium. Another type of embryo that has proven difficult to culture is the parthenogenetic (PN) mouse embryo. With this in mind we have investigated the effect of glucose deprivation on PN embryo development in vitro. Haploid and diploid PN embryos were grown in medium M16 with or without glucose (M16-G) and development, glycolytic rate, and methionine incorporation rates assessed. Haploid PN and normal embryo development to the blastocyst stage did not differ in either M16 or M16-G. In contrast, although diploid PN embryos formed blastocysts in M16 (28.3%), they had difficulty in undergoing the morula/blastocyst transition in M16-G (7.6%). There was no significant difference in mean cell numbers of haploid PN, diploid PN and normal embryos cultured in M16 and M16-G at the morula and blastocyst stage. Transfer of diploid PN embryos from M16-G to M16 at the four- to eight-cell stage dramatically increased blastocyst development. At the morula stage diploid PN embryos grown in M16-G exhibited a higher glucose metabolism and protein synthesis compared to those grown in M16 and to haploid PN embryos. Difficulties of diploid PN embryos in undergoing the morula/blastocyst transition in absence of glucose infer the existence of a link between the maternally inherited components and the preimplantation embryos dependence on glucose.
小鼠植入前胚胎发育的特点是,从致密化前对三羧酸循环的依赖转变为致密化后基于糖酵解的代谢。鉴于此,葡萄糖在胚胎培养基中的作用受到了更多分析,并已导致远交系小鼠胚胎在无葡萄糖培养基中的发育得到改善。另一种已被证明难以培养的胚胎类型是孤雌生殖(PN)小鼠胚胎。考虑到这一点,我们研究了葡萄糖剥夺对PN胚胎体外发育的影响。将单倍体和二倍体PN胚胎在含有或不含有葡萄糖的M16培养基(M16-G)中培养,并评估其发育、糖酵解速率和蛋氨酸掺入率。单倍体PN胚胎和正常胚胎在M16或M16-G中发育到囊胚阶段没有差异。相比之下,虽然二倍体PN胚胎在M16中形成了囊胚(28.3%),但它们在M16-G中进行桑椹胚/囊胚转变时存在困难(7.6%)。在桑椹胚和囊胚阶段,在M16和M16-G中培养的单倍体PN胚胎、二倍体PN胚胎和正常胚胎的平均细胞数没有显著差异。在四细胞到八细胞阶段将二倍体PN胚胎从M16-G转移到M16显著增加了囊胚发育。在桑椹胚阶段,与在M16中培养的二倍体PN胚胎以及单倍体PN胚胎相比,在M16-G中培养的二倍体PN胚胎表现出更高的葡萄糖代谢和蛋白质合成。二倍体PN胚胎在无葡萄糖的情况下进行桑椹胚/囊胚转变存在困难,这意味着母系遗传成分与植入前胚胎对葡萄糖的依赖之间存在联系。