Harris Sarah E, Gopichandran Nadia, Picton Helen M, Leese Henry J, Orsi Nicolas M
Reproduction and Early Development Research Group, School of Medicine, D Floor, Clarendon Wing, Leeds General Infirmary, Belmont Grove, Leeds LS2 9NS, UK.
Theriogenology. 2005 Sep 1;64(4):992-1006. doi: 10.1016/j.theriogenology.2005.01.004.
The culture of murine oocytes and preimplantation embryos in vitro has been used successfully for many years. However, this practice can result in cellular stress and reduced viability. Since this phenomenon is partly attributable to differences in nutrient composition between culture media and maternal tract fluids, we determined the concentrations of glucose, pyruvate, lactate and 19 amino acids in murine preovulatory follicles and oestrous oviductal and uterine fluids. Follicular fluids were aspirated from hyperstimulated ovaries, whereas oviductal fluids (with/without oocyte-cumulus complexes) and uterine fluids were collected from naturally cycling animals. Glucose, pyruvate and lactate concentrations were analysed using ultramicrofluorometric methods, whilst amino acid profiles were determined by reverse-phase high performance liquid chromatography. Mean glucose concentrations in follicular, oviduct (with/without cumulus cells) and uterine fluids were 0.46, 1.09/1.65 and 0.61 mmol l(-1), respectively. Pyruvate concentrations were 0.38, 0.37/0.17 and 0.25 mmol l(-1), respectively, and lactate concentrations were 17.34, 10.92/11.68 and 9.41 mmol l(-1), respectively. Oviductal pyruvate concentration was significantly higher, and glucose significantly lower, in the presence of cumulus cells. Taurine, glycine, alanine, glutamine and glutamate were the major amino acids detected. Concentrations of amino acids differed among fluids, with highest levels being found in the oviduct. The follicular fluid and tract nutrient profiles differed from those of murine maturation, fertilisation and embryo culture media. These data extend our understanding of cellular metabolism and of nutritional environments of the oocyte and early embryo as they progress along the reproductive tract in vivo. These results may also contribute to the formulation of nutritionally more physiological media for mouse oocyte maturation and embryo culture.
多年来，小鼠卵母细胞和植入前胚胎的体外培养已成功应用。然而，这种做法可能导致细胞应激并降低活力。由于这种现象部分归因于培养基和母体生殖道液体之间营养成分的差异，我们测定了小鼠排卵前卵泡、发情期输卵管和子宫液中葡萄糖、丙酮酸、乳酸和19种氨基酸的浓度。从超刺激的卵巢中抽吸卵泡液，而输卵管液（有/无卵母细胞-卵丘复合体）和子宫液则从自然发情的动物中收集。使用超微量荧光法分析葡萄糖、丙酮酸和乳酸浓度，而氨基酸谱则通过反相高效液相色谱法测定。卵泡液、输卵管（有/无卵丘细胞）和子宫液中的平均葡萄糖浓度分别为0.46、1.09/1.65和0.61 mmol l(-1)。丙酮酸浓度分别为0.38、0.37/0.17和0.25 mmol l(-1)，乳酸浓度分别为17.34、10.92/11.68和9.41 mmol l(-1)。在有卵丘细胞的情况下，输卵管丙酮酸浓度显著更高，而葡萄糖浓度显著更低。检测到的主要氨基酸是牛磺酸、甘氨酸、丙氨酸、谷氨酰胺和谷氨酸。不同液体中的氨基酸浓度不同，输卵管中的浓度最高。卵泡液和生殖道营养成分与小鼠成熟、受精和胚胎培养基不同。这些数据扩展了我们对细胞代谢以及卵母细胞和早期胚胎在体内沿生殖道发育过程中营养环境的理解。这些结果也可能有助于制定营养上更符合生理的小鼠卵母细胞成熟和胚胎培养基。
