Institute of Farm Animal Genetics, Friedrich-Loeffler-Institut, Neustadt, Germany.
PLoS One. 2013 Sep 20;8(9):e74981. doi: 10.1371/journal.pone.0074981. eCollection 2013.
Alteration of maternal lipid metabolism early in development has been shown to trigger obesity, insulin resistance, type 2 diabetes and cardiovascular diseases later in life in humans and animal models. Here, we set out to determine (i) lipid composition dynamics in single oocytes and preimplantation embryos by high mass resolution desorption electrospray ionization mass spectrometry (DESI-MS), using the bovine species as biological model, (ii) the metabolically most relevant lipid compounds by multivariate data analysis and (iii) lipid upstream metabolism by quantitative real-time PCR (qRT-PCR) analysis of several target genes (ACAT1, CPT 1b, FASN, SREBP1 and SCAP). Bovine oocytes and blastocysts were individually analyzed by DESI-MS in both positive and negative ion modes, without lipid extraction and under ambient conditions, and were profiled for free fatty acids (FFA), phospholipids (PL), cholesterol-related molecules, and triacylglycerols (TAG). Principal component analysis (PCA) and linear discriminant analysis (LDA), performed for the first time on DESI-MS fused data, allowed unequivocal discrimination between oocytes and blastocysts based on specific lipid profiles. This analytical approach resulted in broad and detailed lipid annotation of single oocytes and blastocysts. Results of DESI-MS and transcript regulation analysis demonstrate that blastocysts produced in vitro and their in vivo counterparts differed significantly in the homeostasis of cholesterol and FFA metabolism. These results should assist in the production of viable and healthy embryos by elucidating in vivo embryonic lipid metabolism.
早期母体脂质代谢的改变已被证明会在人类和动物模型中引发肥胖、胰岛素抵抗、2 型糖尿病和心血管疾病。在这里，我们旨在确定 (i) 通过高分辨率解吸电喷雾电离质谱 (DESI-MS) 分析单个卵母细胞和胚胎前体中脂质组成的动态变化，以牛物种作为生物模型，(ii) 通过多元数据分析确定最相关的代谢脂质化合物，以及 (iii) 通过定量实时 PCR (qRT-PCR) 分析几个靶基因 (ACAT1、CPT 1b、FASN、SREBP1 和 SCAP) 分析脂质上游代谢。牛卵母细胞和囊胚分别通过 DESI-MS 在正离子和负离子模式下进行分析，无需进行脂质提取，且在环境条件下进行，同时对游离脂肪酸 (FFA)、磷脂 (PL)、胆固醇相关分子和三酰基甘油 (TAG) 进行了分析。首次在 DESI-MS 融合数据上进行主成分分析 (PCA) 和线性判别分析 (LDA)，能够基于特定的脂质谱对卵母细胞和囊胚进行明确区分。这种分析方法实现了对单个卵母细胞和囊胚的广泛和详细的脂质注释。DESI-MS 结果和转录调控分析表明，体外培养的囊胚及其体内对应物在胆固醇和 FFA 代谢的动态平衡方面存在显著差异。这些结果应有助于阐明体内胚胎脂质代谢，从而生产出具有活力和健康的胚胎。
