Leão Beatriz C S, Rocha-Frigoni Nathália A S, Cabral Elaine C, Franco Marcos F, Ferreira Christina R, Eberlin Marcos N, Filgueiras Paulo R, Mingoti Gisele Z
School of Veterinary Medicine,Department of Animal Health,UNESP-Univ Estadual Paulista,Rua Clóvis Pestana 793,16050-680,Araçatuba,São Paulo,Brazil.
ThoMSon Mass Spectrometry Laboratory,Chemistry Institute,University of Campinas (UNICAMP),Cidade Universitária Zeferino Vaz s/n,CP 6154,bloco A6,sala 111,13083-970,Distrito de Barão Geraldo-Campinas,São Paulo,Brazil.
Zygote. 2015 Oct;23(5):732-41. doi: 10.1017/S0967199414000380. Epub 2014 Sep 12.
This study aimed to evaluate the impact of vitrification on membrane lipid profile obtained by mass spectrometry (MS) of in vitro-produced bovine embryos. Matrix-assisted laser desorption ionization-mass spectrometry (MALDI-MS) has been used to obtain individual embryo membrane lipid profiles. Due to conditions of analysis, mainly membrane lipids, most favorably phosphatidylcholines (PCs) and sphingomyelins (SMs) have been detected. The following ions described by their mass-to-charge ratio (m/z) and respective attribution presented increased relative abundance (1.2-20×) in the vitrified group: 703.5 [SM (16:0) + H]+; 722.5 [PC (40:3) + Na]+; 758.5 [PC (34:2) + H]+; 762.5 [PC (34:0) + H]+; 790.5 [PC (36:0) + H]+ and 810.5 [PC (38:4) + H]+ and/or [PC (36:1) + Na]+. The ion with a m/z 744.5 [PCp (34:1) and/or PCe (34:2)] was 3.4-fold more abundant in the fresh group. Interestingly, ions with m/z 722.5 or 744.5 indicate the presence of lipid species, which are more resistant to enzymatic degradation as they contain fatty acyl residues linked through ether type bonds (alkyl ether or plasmalogens, indicated by the lowercase 'e' and 'p', respectively) to the glycerol structure. The results indicate that cryopreservation impacts the membrane lipid profile, and that these alterations can be properly monitored by MALDI-MS. Membrane lipids can therefore be evaluated by MALDI-MS to monitor the effect of cryopreservation on membrane lipids, and to investigate changes in lipid profile that may reflect the metabolic response to the cryopreservation stress or changes in the environmental conditions.
本研究旨在评估玻璃化冷冻对体外生产的牛胚胎通过质谱(MS)获得的膜脂质谱的影响。基质辅助激光解吸电离质谱(MALDI-MS)已被用于获取单个胚胎的膜脂质谱。由于分析条件,主要是膜脂质,最有利地检测到了磷脂酰胆碱(PCs)和鞘磷脂(SMs)。以下按质荷比(m/z)描述并给出各自归属的离子在玻璃化冷冻组中相对丰度增加(1.2 - 20倍):703.5 [SM (16:0) + H]+;722.5 [PC (40:3) + Na]+;758.5 [PC (34:2) + H]+;762.5 [PC (34:0) + H]+;790.5 [PC (36:0) + H]+ 和810.5 [PC (38:4) + H]+ 和/或 [PC (36:1) + Na]+。质荷比为744.5 [PCp (34:1) 和/或PCe (34:2)] 的离子在新鲜组中的丰度高3.4倍。有趣的是,质荷比为722.5或744.5的离子表明存在脂质种类,它们对酶促降解更具抗性,因为它们含有通过醚型键(烷基醚或缩醛磷脂,分别由小写字母“e”和“p”表示)与甘油结构相连的脂肪酰基残基。结果表明冷冻保存会影响膜脂质谱,并且这些变化可以通过MALDI-MS进行适当监测。因此,可以通过MALDI-MS评估膜脂质,以监测冷冻保存对膜脂质的影响,并研究可能反映对冷冻保存应激的代谢反应或环境条件变化的脂质谱变化。