Analysis of mitochondrial function in mammalian oocytes using electrochemical microscopy

耗氧量是一个普遍存在的参数，可以提供有价值的信息代谢机制，卵母细胞和胚胎质量。本研究的目的是：1）建立一种基于扫描电化学显微镜（SECM）的单卵母细胞和胚胎细胞呼吸测量装置; 2）研究不同发育阶段的单卵母细胞和胚胎线粒体呼吸功能，如耗氧量和细胞色素氧化酶（考克斯）基因表达。我们成功地研制了一种改进的SECM测量系统，包括铂微盘电极和呼吸测定介质，可以监测单个卵母细胞和胚胎的耗氧率。利用该系统，我们可以对不同发育阶段的牛和小鼠的单个胚胎和卵母细胞进行呼吸活性的测定。线粒体呼吸活性的发展，如线粒体的膜电位和考克斯的基因表达，对应于胚胎发育过程中耗氧速率的增加。

Oxygen consumption is a ubiquitous parameter which can provide valuable informationon metabolic mechanisms, and oocyte and embryo quality. The aim of this study was 1) todevelop a device to measure cellular respiration of single oocytes and embryos based onscanning electrochemical microscopy (SECM) and 2) to assess the mitochondrial respiration function, such as oxygen consumption and gene expression of cytochrome coxidase (Cox) in single oocytes and embryos at different developmental stages. We succeeded in development of a modified SECM measuring system including Pt-microdiskelectrodes and respiration assay medium can monitor the oxygen consumption rate ofsingle oocytes and embryos. Using this system, we can measure respiration activity bysingle embryos and oocytes of bovine and mice at different developmental stages. Development of mitochondrial respiration activity, such as membrane potential of mitochondria and gene expression of Cox, corresponds to the increase of oxygen consumption rates during the development of embryos.