Baroffio A, Kucera P
J Cell Physiol. 1985 Apr;123(1):111-6. doi: 10.1002/jcp.1041230116.
14C-2-deoxyglucose (DG), currently employed in in vivo studies of brain glucose metabolism, has been used for determination of glucose consumption in the in vitro developing chick embryo. DG, presented in traces, accumulates in the embryo in proportion with incubation time. Analysis of tissue homogenates shows that the accumulated radioactivity is due to both phosphorylated (DGP) and nonphosphorylated DG. As it is only the radioactivity originating from the DGP that is proportional to glucose utilization, the nonphosphorylated DG must be washed out. The washout shows two distinct kinetics: a fast one corresponding to DG that has entered the cells but has not yet been phosphorylated and a slow one that is probably due to a dephosphorylated DGP coming from a different cellular compartment. On the basis of these results the optimal experimental conditions have been defined, allowing quantitative studies of glucose metabolism during the first day of development of the chicken embryo. From 18 to 24 hr of incubation (end of gastrulation), total glucose consumption increases from 50 nmol X h-1 at stage 3-4 to 90 nmol X h-1 at stage 6-7. This increase mainly reflects the growth of the blastodisc. Comparison with the values of O2 uptake measured at the same period of development suggests that only a fraction of the glucose consumed is oxidized, the major part being converted aerobically to lactate.
14C - 2 - 脱氧葡萄糖(DG)目前用于脑葡萄糖代谢的体内研究,也已被用于测定体外发育的鸡胚中的葡萄糖消耗。痕量存在的DG在胚胎中随孵化时间成比例积累。对组织匀浆的分析表明,积累的放射性是由于磷酸化的(DGP)和未磷酸化的DG两者所致。由于只有源自DGP的放射性与葡萄糖利用成比例,所以必须洗去未磷酸化的DG。洗脱显示出两种不同的动力学:一种快速的对应于已进入细胞但尚未磷酸化的DG,另一种缓慢的可能是由于来自不同细胞区室的去磷酸化DGP。基于这些结果,确定了最佳实验条件,从而能够对鸡胚发育第一天期间的葡萄糖代谢进行定量研究。在孵化18至24小时(原肠胚形成末期),总葡萄糖消耗从第3 - 4阶段的50 nmol×h-1增加到第6 - 7阶段的90 nmol×h-1。这种增加主要反映了胚盘的生长。与在相同发育时期测得的氧气摄取值比较表明,消耗的葡萄糖只有一部分被氧化,大部分通过有氧途径转化为乳酸。