Complete spatial characterisation of neutral and sialylated N-glycans in an adult rodent brain Juhi Samal1, Radka Saldova2, Pauline M.Rudd2, Róisín O'Flaherty2, Abhay Pandit1 1Center for Research in Medical Devices (CÚRAM), NUI Galway; 2National Institute for Bioprocessing Research and Training (NIBRT), UCD, Ireland
The cardinal role of N-glycans for the nervous system is brought forth by glycosylation alterations, resulting in different neuropathological symptoms like mental retardation, seizures and epilepsy. Previous studies have reported the conservation of N-glycan processing in rodents1. However, there is a lack of spatial resolution of these glycan profiles as the tissue samples were homogenized intact for these studies or specific proteins were picked up for analysis of glycosylation. Since the biosynthesis of N-glycans undergoes a stringent spatio-temporal regulation within the tissue, we hypothesize that the spatial resolution of N-glycans isolated from striatum and substantia nigra (SN) can give an insight into their involvement in regulation of striatal and nigral cues for establishment and pathophysiological degeneration of neural circuitry in Parkinson's disease. Specific objectives of the study involve isolation of N-glycans from rat striatum and SN, reproducibility, resolution and relative quantitation of N-glycome using ultra performance liquid chromatography (UPLC), weak anion exchange UPLC (WAX-UPLC), liquid chromatography mass spectrometry (LC-MS), lectin array and lectin histochemistry. Whole N-glycome from striatum and SN (GP1-GP26 as observed in UPLC profile) were characterized using exoglycosidase digestions and LC-MS. It revealed significant differences in sialylation patterns of striatum and SN N-glycans; with more pronounced mono- (striatum: 8.46%,SN: 7.16%) and tri-sialylation (striatum: 4.54%,SN: 3.44%) in striatal N-glycans. Derived glycosylation traits like polysialic acid content were also calculated for N-glycans with same structural features using the ratio of glycan peaks in UPLC. This study presents a detailed comparative spatial analysis of relative abundances of N- glycans in striatum and SN of rodent brains, serving as a foundation for identifying the 'brain-type' glycans as biomarkers or therapeutic targets and their modulation in neurodegenerative disorders.
Reference: 1Zamze, S. et al.Eur. J. Biochem. 1998 Hardiman Fellowship, NUI Galway and Science Foundation Ireland and European Regional Development Fund (13/RC/2073)
Credits: None available.
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