title: Neuronal glycosylation differentials in normal, injured and chondroitinase-treated environments.
abstract: Glycosylation is found ubiquitously throughout the central nervous system (CNS). Chondroitin sulphate proteoglycans (CSPGs) are a group of molecules heavily substituted with glycosaminoglycans (GAGs) and are found in the extracellular matrix (ECM) and cell surfaces. Upon CNS injury, a glial scar is formed, which is inhibitory for axon regeneration. Several CSPGs are up-regulated within the glial scar, including NG2, and these CSPGs are key inhibitory molecules of axonal regeneration. Treatment with chondroitinase ABC (ChABC) can neutralise the inhibitory nature of NG2. A gene expression dataset was mined in silico to verify differentially regulated glycosylation-related genes in neurons after spinal cord injury and identify potential targets for further investigation. To establish the glycosylation differential of neurons that grow in a healthy, inhibitory and ChABC-treated environment, we established an indirect co-culture system where PC12 neurons were grown with primary astrocytes, Neu7 astrocytes (which overexpress NG2) and Neu7 astrocytes treated with ChABC. After 1, 4 and 8 days culture, lectin cytochemistry of the neurons was performed using five fluorescently-labelled lectins (ECA MAA, PNA, SNA-I and WFA). Usually α-(2,6)-linked sialylation scarcely occurs in the CNS but this motif was observed on the neurons in the injured environment only at day 8. Treatment with ChABC was successful in returning neuronal glycosylation to normal conditions at all timepoints for MAA, PNA and SNA-I staining, and by day 8 in the case of WFA. This study demonstrated neuronal cell surface glycosylation changes in an inhibitory environment and indicated a return to normal glycosylation after treatment with ChABC, which may be promising for identifying potential therapies for neuronal regeneration strategies.
authors.affiliation: Glycoscience Group, National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
authors.forename: Michelle
authors.initials: M
authors.lastname: Kilcoyne
authors.forename: Shashank
authors.initials: S
authors.lastname: Sharma
authors.forename: Niamh
authors.initials: N
authors.lastname: McDevitt
authors.forename: Claire
authors.initials: C
authors.lastname: O'Leary
authors.forename: Lokesh
authors.initials: L
authors.lastname: Joshi
authors.forename: Siobhán S
authors.initials: SS
authors.lastname: McMahon
mesh_terms: D000818:Animals; D000941:Antigens / Q000494:pharmacology; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000494:pharmacology*; D018920:Coculture Techniques; D006031:Glycosylation; D037102:Lectins / Q000235:genetics; D009416:Nerve Regeneration / Q000187:drug effects*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology*; D016716:PC12 Cells; D011509:Proteoglycans / Q000037:antagonists & inhibitors / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013119:Spinal Cord Injuries / Q000378:metabolism / Q000473:pathology / Q000503:physiopathology; D000099216:Chondroitin Sulfate Proteoglycan 4
chemical_list: D000941:Antigens; D019765:Chondroitin ABC Lyase; D037102:Lectins; D011509:Proteoglycans; D000099216:Chondroitin Sulfate Proteoglycan 4