earticle

영어

Gangliosides are amomc glycosphingolipids containing one to several sialic acid residues. Although they play an important role in neuro-biological functions including synaptic plasticity and memory formation, there are only few studies in ganglioside due to their structural complexity and lack of effective analytical methods. In particular, isomeric investigation of gangliosides is necessarily required in order to understand the ganglioside biology with the consideration of its biosynthesis however, isomer separation based on reversed-phase (RP) chromatography is highly challengeable. In this study, for the purpose of construction of localized mouse brain ganglioside library, we examined ganglioside from anatomically dissected nine mouse brain regions using UHPLC-QTOF MS and MS/MS. Briefly, gangliosides were extracted by modified Folch method with chloroform and methanol, followed by purification and enrichment by Cl8-SPE. Then they were identified and quantified by UHPLC (Cl8 column) QTOF MS. Ganglioside isomers were completely separated depending on their glycan traits on a Cl8 column by addition of formic acid in the mobile phase. Furthermore, tandem MS analysis was performed to confirm the identification based on diagnostic fragment ions. Through LC-MS/MS based isomeric investigation of gangliosides, region- specific mouse brain ganglioside biosynthetic pathway including 0-, a-, b- and c- series could be suggested. Interestingly, major ganglioside were distributed with distinguished qualitative and quantitative pattern for each nine brain regions. In the future, the constructed library of mouse brain ganglioside will be used for monitoring alteration in KO mouse models including glycosylation transferase KO mice.