INRA, UMR85 Physiologie de la Reproduction et des Comportements, Équipe Interactions Cellulaires et Fertilité, 37380 Nouzilly, France.
Reproduction. 2013 Jul 31;146(3):221-31. doi: 10.1530/REP-13-0142. Print 2013 Sep.
The microenvironment of the developing follicle is critical to the acquisition of oocyte developmental competence, which is influenced by several factors including follicle size and season. The aim of this study was to characterise the metabolomic signatures of porcine follicular fluid (FF) collected from good and poor follicular environments, using high-resolution proton nuclear magnetic resonance (1H-NMR) spectroscopy. Sow ovaries were collected at slaughter, 4 days after weaning, in summer and winter. The contents of small (3-4 mm) and large (5-8 mm) diameter follicles were aspirated and pooled separately for each ovary pair. Groups classified as summer-small (n=8), summer-large (n=15), winter-small (n=9) and winter-large (n=15) were analysed by 1H-NMR spectroscopy. The concentrations of 11 metabolites differed due to follicle size alone (P<0.05), including glucose, lactate, hypoxanthine and five amino acids. The concentrations of all these metabolites, except for glucose, were lower in large FF compared with small FF. Significant interaction effects of follicle size and season were found for the concentrations of glutamate, glycine, N-acetyl groups and uridine. Succinate was the only metabolite that differed in concentration due to season alone (P<0.05). The FF levels of progesterone, androstenedione and oestradiol were correlated with the concentrations of most of the metabolites examined. The results indicate that there is a distinct shift in follicular glucose metabolism as follicles increase in diameter and suggest that follicular cells may be more vulnerable to oxidative stress during the summer months. Our findings demonstrate the power of 1H-NMR spectroscopy to expand our understanding of the dynamic and complex microenvironment of the developing follicle.
猪卵泡液的代谢组学特征分析,使用高分辨率质子核磁共振(1H-NMR)光谱法
猪卵泡液(FF)的微环境对卵母细胞发育能力的获得至关重要,这受到许多因素的影响,包括卵泡大小和季节。本研究旨在使用高分辨率质子核磁共振(1H-NMR)光谱法,分析来自良好和不良卵泡微环境的猪卵泡液(FF)的代谢组学特征。在屠宰时、断奶后 4 天、夏季和冬季采集母猪的卵巢。从小(3-4mm)和大(5-8mm)直径卵泡中抽吸内容物,并分别对每对卵巢进行分组。根据 1H-NMR 光谱分析,将夏季小卵泡(n=8)、夏季大卵泡(n=15)、冬季小卵泡(n=9)和冬季大卵泡(n=15)分组。结果表明,仅因卵泡大小的不同就导致 11 种代谢物的浓度存在差异(P<0.05),其中包括葡萄糖、乳酸、次黄嘌呤和 5 种氨基酸。与小卵泡 FF 相比,大卵泡 FF 中的所有这些代谢物(除葡萄糖外)的浓度均较低。还发现了卵泡大小和季节对谷氨酸、甘氨酸、N-乙酰基和尿苷浓度的显著互作效应。仅因季节的不同就导致琥珀酸的浓度存在差异(P<0.05)。FF 中孕酮、雄烯二酮和雌二醇的水平与所检测到的大多数代谢物的浓度相关。结果表明,随着卵泡直径的增加,卵泡的葡萄糖代谢发生了明显的变化,并提示在夏季,卵泡细胞可能更容易受到氧化应激的影响。本研究结果表明,1H-NMR 光谱法具有强大的功能,可以扩展我们对发育中卵泡动态复杂微环境的理解。