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Orbitrap mass spectrometry for monitoring the ganglioside pattern in human cerebellum development and aging
Authors:Raluca Ica  Alina Petrut  Cristian VA Munteanu  Mirela Sarbu   eljka Vukeli&#x;  Ligia Petrica  Alina D Zamfir
Institution:Raluca Ica,Alina Petrut,Cristian V.A. Munteanu,Mirela Sarbu,Željka Vukelić,Ligia Petrica,Alina D. Zamfir
Abstract: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 CH3COO? 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.
Keywords:cerebellum  gangliosides  high resolution  Orbitrap mass spectrometry  sequencing  screening  structural analysis
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