earticle

영어

Glycan plays numerous roles in the nervous system during development, regeneration and synaptic plasticity, forming of a complex meshwork at the cell surface. We newly established an analytical platform for exploring two molecules- glycan attached to proteins and glycan attached to lipids- from human brain using mass spectrometry. 69 brain samples were obtained from the University of Cambridge. Glycan extraction method from previous study, and new ganglioside extraction method were paired with MS. Briefly, each brain tissues were grinded by sonication. Then for glycans, the membrane was pelleted by ultracentrifugation. N-glycan release and associated processing steps were performed by previously optimized procedures. For sphingolipids, homogenized brain is mixed with chloroform/methanol/water and centrifuged. Total lipids extracts were partitioned again with chloroform/methanol to collect gangliosides. After further enrichment by SPE, N-glycans and gangliosides were profiled by positive and negative ion mode nano-LC/MS respectively. Stucture information was obtained by nano-LC/MS/MS. Brain samples were divided into the 7 groups: neonates, infants, toddlers, school-ages, teenagers, young adults. We successfully profiled brain glycome and sphingolipids, identifying approximately 100 N-glycan compositions and 120 ganglioside compounds with several unique “brain type” characters. This study promise insights into involvement of glycans to massive flow of information that underpins brain, with providing meaningful understanding to neuropathology related with aging.