Thomson Mass Spectrometry Laboratory, University of Campinas, Campinas, SP, Brazil.
J Lipid Res. 2010 May;51(5):1218-27. doi: 10.1194/jlr.D001768. Epub 2009 Nov 5.
Methods used for lipid analysis in embryos and oocytes usually involve selective lipid extraction from a pool of many samples followed by chemical manipulation, separation and characterization of individual components by chromatographic techniques. Herein we report direct analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of single and intact embryos or oocytes from various species. Biological samples were simply moisturized with the matrix solution and characteristic lipid (represented by phosphatidylcholines, sphingomyelins and triacylglycerols) profiles were obtained via MALDI-MS. As representative examples, human, bovine, sheep and fish oocytes, as well as bovine and insect embryos were analyzed. MALDI-MS is shown to be capable of providing characteristic lipid profiles of gametes and embryos and also to respond to modifications due to developmental stages and in vitro culture conditions of bovine embryos. Investigation in developmental biology of the biological roles of structural and reserve lipids in embryos and oocytes should therefore benefit from these rapid MALDI-MS profiles from single and intact species.
用于胚胎和卵母细胞脂质分析的方法通常涉及从许多样品池中选择性地提取脂质,然后通过色谱技术对单个成分进行化学处理、分离和特征化。本文报告了一种通过基质辅助激光解吸/电离质谱(MALDI-MS)直接分析来自不同物种的单个完整胚胎或卵母细胞的方法。生物样品只需用基质溶液润湿,然后通过 MALDI-MS 即可获得特征脂质(以磷脂酰胆碱、神经鞘磷脂和三酰基甘油表示)图谱。作为代表性实例,分析了人、牛、绵羊和鱼类卵母细胞以及牛和昆虫胚胎。结果表明,MALDI-MS 能够提供配子和胚胎的特征脂质图谱,并且还能够响应牛胚胎发育阶段和体外培养条件的变化。因此,这些来自单个完整物种的快速 MALDI-MS 图谱应该有助于胚胎和卵母细胞中结构和储备脂质的生物学作用的发育生物学研究。