Desorption electrospray ionization mass spectrometry reveals lipid metabolism of individual oocytes and embryos.
Desorption electrospray ionization mass spectrometry reveals lipid metabolism of individual oocytes and embryos.
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DOI：
10.1371/journal.pone.0074981
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Cooks RG
中科院分区：
文献类型：
--
作者：
González-Serrano AF;Pirro V;Ferreira CR;Oliveri P;Eberlin LS;Heinzmann J;Lucas-Hahn A;Niemann H;Cooks RG
在人类和动物模型中，发育早期母体脂质代谢的改变已被证明会在生命后期引发肥胖、胰岛素抵抗、2型糖尿病和心血管疾病。在这里，我们以牛为生物模型，通过高质量分辨率解吸电喷雾电离质谱（DESI-MS）确定(i)单个卵母细胞和植入前胚胎的脂质组成动态，（ii）通过多变量数据分析确定代谢最相关的脂质化合物，（iii）通过定量实时PCR （qRT-PCR）分析几个靶基因（ACAT1, CPT 1b， FASN， SREBP1和SCAP）的脂质上游代谢。牛卵母细胞和囊胚分别在正离子和负离子模式下进行DESI-MS分析，不进行脂质提取，并在环境条件下进行游离脂肪酸（FFA）、磷脂（PL）、胆固醇相关分子和三酰甘油（TAG）的分析。首次对DESI-MS融合数据进行主成分分析（PCA）和线性判别分析（LDA），可以根据特定的脂质谱明确区分卵母细胞和囊胚。这种分析方法对单个卵母细胞和囊胚进行了广泛而详细的脂质注释。DESI-MS和转录调控分析结果表明，体外产生的囊胚与体内囊胚在胆固醇和游离脂肪酸代谢的稳态方面存在显著差异。这些结果应该有助于通过阐明体内胚胎脂质代谢来产生有活力和健康的胚胎。
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.
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影响因子：
4.3
作者：
Ferreira, Christina R.;Pirro, Valentina;Cooks, R. Graham
通讯作者：
Cooks, R. Graham
影响因子：
4.4
作者：
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通讯作者：
Khatib H
影响因子：
3.6
作者：
Gad, Ahmed;Hoelker, Michael;Tesfaye, Dawit
通讯作者：
Tesfaye, Dawit
影响因子：
6.5
作者：
Ferreira, Christina R.;Saraiva, Sergio A.;Eberlin, Marcos N.
通讯作者：
Eberlin, Marcos N.
影响因子：
7.4
作者：
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通讯作者：
Wiseman, Justin M.