Department of Electrical and Computer Engineering, Colorado State University, Fort Collins, CO 80523, USA.
Department of Biomedical Sciences, Colorado State University, Fort Collins, CO 80523, USA.
Biosens Bioelectron. 2019 Feb 1;126:615-623. doi: 10.1016/j.bios.2018.09.071. Epub 2018 Sep 21.
This paper presents the development of a multi-sensor platform capable of simultaneous measurement of dissolved oxygen (DO) concentration, glucose and lactate concentrations in a micro-chamber for real-time evaluation of metabolic flux in bovine embryos. A micro-chamber containing all three sensors (DO, glucose, and lactate) was made to evaluate metabolic flux of single oocytes or embryos at different stages of development in ≤ 120 µL of respiration buffer. The ability of the sensor to detect a metabolic shift from oxidative phosphorylation (OXPHOS) to glycolysis was demonstrated in embryos by an ablation of oxygen consumption and an increase in lactate production following addition of oligomycin, an inhibitor of mitochondrial adenosine triphosphate (ATP) synthesis. An increased reliance upon glycolysis relative to OXPHOS was demonstrated in embryos as they developed from morula to hatched blastocysts by a progressive increase in the lactate/oxygen flux ratio, consistent with isolated metabolic assessments reported previously. These studies highlight the utility of a metabolic multi-sensor for integrative real-time monitoring of aerobic and anaerobic energy metabolism in bovine embryos, with potential applications in the study of metabolic processes in oocyte and early embryonic development.
本文介绍了一种多传感器平台的开发,该平台能够同时测量微室中的溶解氧 (DO) 浓度、葡萄糖和乳酸浓度,以便实时评估牛胚胎中的代谢通量。制作了一个包含所有三个传感器(DO、葡萄糖和乳酸)的微室,以评估在 ≤ 120 µL 呼吸缓冲液中处于不同发育阶段的单个卵母细胞或胚胎的代谢通量。通过添加寡霉素(一种抑制线粒体三磷酸腺苷 (ATP) 合成的抑制剂)来消除氧消耗并增加乳酸产生,证明了传感器能够检测从氧化磷酸化 (OXPHOS) 到糖酵解的代谢转变。随着胚胎从桑葚胚发育到孵出的囊胚,糖酵解相对于 OXPHOS 的依赖性增加,这与以前报道的分离代谢评估一致,通过乳酸/氧通量比的逐渐增加来证明。这些研究强调了代谢多传感器在整合实时监测牛胚胎有氧和无氧能量代谢方面的实用性,在卵母细胞和早期胚胎发育代谢过程的研究中有潜在的应用。