Pirro Valentina, Oliveri Paolo, Ferreira Christina Ramires, González-Serrano Andrés Felipe, Machaty Zoltan, Cooks Robert Graham
Department of Chemistry, University of Turin, Via Pietro Giuria 7, Turin 10125, Italy.
Department of Pharmacy, University of Genoa, Via Brigata Salerno 13, Genoa 16147, Italy.
Anal Chim Acta. 2014 Oct 27;848:51-60. doi: 10.1016/j.aca.2014.08.001. Epub 2014 Aug 11.
The development of sensitive measurements to analyze individual cells is of relevance to elucidate specialized roles or metabolic functions of each cell under physiological and pathological conditions. Lipids play multiple and critical roles in cellular functions and the application of analytical methods in the lipidomics area is of increasing interest. In this work, in vitro maturation of porcine oocytes was studied. Two independent sources of chemical information (represented by mass spectra in the positive and negative ion modes) from single oocytes (immature oocytes, 24-h and 44-h in vitro matured oocytes) were acquired by using desorption electrospray ionization-mass spectrometry (DESI-MS). Low and mid-level data fusion strategies are presented with the aim of better exploring the large amount of chemical information contained in the two mass spectrometric lipid profiles. Data were explored by principal component analysis (PCA) within the two multi-block approaches to include information on free fatty acids, phospholipids, cholesterol-related molecules, di- and triacylglycerols. After data fusion, clearer differences among immature and in vitro matured porcine oocytes were observed, which provide novel information regarding lipid metabolism throughout oocyte maturation. In particular, changes in TAG composition, as well as increase in fatty acid metabolism and membrane complexity were evidenced during the in vitro maturation process. This information can assist the improvement of in vitro embryo production for porcine species.
开发用于分析单个细胞的灵敏测量方法对于阐明每个细胞在生理和病理条件下的特殊作用或代谢功能具有重要意义。脂质在细胞功能中发挥着多种关键作用,脂质组学领域分析方法的应用越来越受到关注。在这项工作中,研究了猪卵母细胞的体外成熟。通过解吸电喷雾电离质谱(DESI-MS)从单个卵母细胞(未成熟卵母细胞、体外成熟24小时和44小时的卵母细胞)获取了两种独立的化学信息来源(以正离子和负离子模式下的质谱表示)。提出了低水平和中等水平的数据融合策略,旨在更好地探索两种质谱脂质谱中包含的大量化学信息。在两种多块方法中通过主成分分析(PCA)对数据进行探索,以纳入有关游离脂肪酸、磷脂、胆固醇相关分子、二酰基甘油和三酰基甘油的信息。数据融合后,观察到未成熟和体外成熟猪卵母细胞之间有更明显的差异,这为整个卵母细胞成熟过程中的脂质代谢提供了新信息。特别是,在体外成熟过程中,三酰基甘油(TAG)组成发生变化,脂肪酸代谢增加,膜复杂性提高。这些信息有助于改进猪的体外胚胎生产。