We have developed here a superior approach based on high-resolution (HR) mass spectrometry (MS) for monitoring the changes occurring with development and aging in the composition and structure of cerebellar gangliosidome. The experiments were focused on the comparative screening and structural analysis of gangliosides expressed in fetal and aged cerebellum by Orbitrap MS with nanoelectrospray ionization (nanoESI) in the negative ion mode. The employed ultrahigh-resolution MS platform allowed the discrimination, without the need of previous separation, of 159 ions corresponding to 120 distinct species in the native ganglioside mixtures from fetal and aged cerebellar biopsies, many more than detected before, when MS platforms of lower resolution were employed. A number of gangliosides, in particular polysialylated belonging to GT, GQ, GP, and GS classes, modified by O-fucosylation, O-acetylation, or CH3 COO- were discovered here, for the first time in human cerebellum. These components, found differently expressed in fetal and aged tissues, indicated that the ganglioside profile in cerebellum is development stage- and age-specific. Following the fragmentation analysis by high-energy collision-induced dissociation (HCD) tandem MS (MS/MS), we have also observed that the intimate structure of certain compounds has not changed during the development and aging of the brain, an aspect which could open new directions in the investigation of ganglioside biomarkers in cerebellar tissue.

中文翻译：

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Orbitrap质谱用于监测人类小脑发育和衰老中神经节苷脂模式。

我们在这里开发了一种基于高分辨率（HR）质谱（MS）的高级方法，用于监视小脑神经节体的组成和结构随着发育和衰老而发生的变化。实验的重点是通过负离子模式的纳米电喷雾电离（nanoESI）的Orbitrap MS对胎儿和老年小脑中表达的神经节苷脂进行比较筛选和结构分析。所采用的超高分辨率MS平台无需先分离即可辨别来自胎儿和老年小脑活检组织的天然神经节苷脂混合物中的120种不同物种的159个离子，这比以前使用低分辨率MS平台时所检测到的要多得多。被雇用。许多神经节苷脂，特别是属于GT，GQ，GP的多唾液酸化，并在人小脑中首次发现了通过O-岩藻糖基化，O-乙酰化或CH3 COO-修饰的GS和GS类。这些成分在胎儿和老年组织中表达不同，表明小脑中神经节苷脂的分布是特定于发育阶段和年龄的。在通过高能碰撞诱导解离（HCD）串联质谱（MS / MS）进行的碎片分析之后，我们还观察到某些化合物的紧密结构在大脑的发育和衰老过程中没有改变，这一方面可以在研究小脑组织中神经节苷脂生物标志物方面开辟了新的方向。指出小脑神经节苷脂谱是特定于发育阶段和年龄的。在通过高能碰撞诱导解离（HCD）串联质谱（MS / MS）进行的碎片分析之后，我们还观察到某些化合物的紧密结构在大脑的发育和衰老过程中没有改变，这一方面可以在研究小脑组织中神经节苷脂生物标志物方面开辟了新的方向。指出小脑神经节苷脂谱是特定于发育阶段和年龄的。在通过高能碰撞诱导解离（HCD）串联质谱（MS / MS）进行的碎片分析之后，我们还观察到某些化合物的紧密结构在大脑的发育和衰老过程中没有改变，这一方面可以在研究小脑组织中神经节苷脂生物标志物方面开辟了新的方向。