Developmental phases of individual mouse preimplantation embryos characterized by lipid signatures using desorption electrospray ionization mass spectrometry
Developmental phases of individual mouse preimplantation embryos characterized by lipid signatures using desorption electrospray ionization mass spectrometry
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DOI：
10.1007/s00216-012-6426-4
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发表时间：
2012-12-01
影响因子：
4.3
通讯作者：
Cooks, R. Graham
中科院分区：
文献类型：
--
作者：
Ferreira, Christina R.;Pirro, Valentina;Cooks, R. Graham
个体植入前胚胎中存在的脂质的知识在胚胎学的基础研究中是有意义的，在尝试理解细胞多能性和优化生物技术（如体外受精和转基因）的应用和发展所必需的体外培养条件中是有意义的。在这项工作中，个人的小鼠植入前胚胎和卵母细胞的脂肪酸和磷脂（PL）的配置文件，获得使用的解析策略的基础上解吸电喷雾质谱（EST-MS）。该方法避免了样品制备，并提供了这些微观结构中存在的脂质的信息。未受精的卵母细胞，两个和四个细胞的胚胎，和囊胚中观察到的脂质谱的差异进行了表征。对于一组具有代表性的胚胎（N = 114），使用多变量分析（特别是主成分分析）未受精卵母细胞显示出较窄的范围内的PL物种比囊胚。与未受精的卵母细胞相比，二细胞和四细胞胚胎显示出广泛的PL和高丰度的脂肪酸，表明明显的合成活性。这些数据表明，在小鼠着床前发育中观察到的脂质变化反映了胚泡阶段细胞膜功能和结构特化程度的获得。还值得注意的是，与体内衍生的对应物相比，从两细胞到胚泡阶段体外培养的胚胎具有更均匀的脂质分布，这归因于合成培养基中存在的营养物质的有限多样性。从脂质生物化学和已知对早期胚胎发育至关重要的不同脂质的基因表达的先前报告中解释了EST-MS数据。
Knowledge of the lipids present in individual preimplantation embryos is of interest in fundamental studies of embryology, in attempts to understand cellular pluripotency and in optimization of in vitro culture conditions necessary for the application and development of biotechnologies such as in vitro fertilization and transgenesis. In this work, the profiles of fatty acids and phospholipids (PL) in individual mouse preimplantation embryos and oocytes were acquired using an analytical strategy based on desorption electrospray ionization mass spectrometry (DESI-MS). The methodology avoids sample preparation and provides information on the lipids present in these microscopic structures. Differences in the lipid profiles observed for unfertilized oocytes, two- and four-cell embryos, and blastocysts were characterized. For a representative set of embryos (N = 114) using multivariate analysis (specifically principal component analysis) unfertilized oocytes showed a narrower range of PL species than did blastocysts. Two- and four-cell embryos showed a wide range of PLs compared with unfertilized oocytes and high abundances of fatty acids, indicating pronounced synthetic activity. The data suggest that the lipid changes observed in mouse preimplantation development reflect acquisition of a degree of cellular membrane functional and structural specialization by the blastocyst stage. It is also noteworthy that embryos cultured in vitro from the two-cell through the blastocyst stage have a more homogeneous lipid profile as compared with their in vivo-derived counterparts, which is ascribed to the restricted diversity of nutrients present in synthetic culture media. The DESI-MS data are interpreted from lipid biochemistry and previous reports on gene expression of diverse lipids known to be vital to early embryonic development.