title: Increased Expression of GM1 Detected by Electrospray Mass Spectrometry in Rat Primary Embryonic Cortical Neurons Exposed to Glutamate Toxicity.
abstract: Neurons within different brain regions have varying levels of vulnerability to external stress and respond differently to injury. A potential reason to explain this may lie within a key lipid class of the cell's plasma membrane called gangliosides. These glycosphingolipid species have been shown to play various roles in the maintenance of neuronal viability. The purpose of this study is to use electrospray ionization mass spectrometry (ESI-MS) and immunohistochemistry to evaluate the temporal expression profiles of gangliosides during the course of neurodegeneration in rat primary cortical neurons exposed to glutamate toxicity. Primary embryonic (E18) rat cortical neurons were cultured to DIV (days in vitro) 14. Glutamate toxicity was induced for 1, 3, 6, and 24 h to injure and kill neurons. Immunofluorescence was used to stain for GM1 and GM3 species, and ESI-MS was used to quantify the ganglioside species expressed within these injured neurons. ESI-MS data revealed that GM1, GM2, and GM3 were up-regulated in neurons exposed to glutamate. Interestingly, using immunofluorescence, we demonstrated that the GM1 increase following glutamate exposure occurred in viable neurons, possibly indicating a potential intrinsic neuroprotective response. To test this potential neuroprotective property, neurons were pretreated with GM1 for 24 h prior to glutamate exposure. Pretreatment with GM1 conferred significant neuroprotection against glutamate-induced cell death. Overall, work from this study validates the use of ESI-MS for cell-derived gangliosides and supports the further development of lipid based strategies to protect against neuron cell death.
authors.forename: Dae Hee
authors.initials: DH
authors.lastname: Park
authors.forename: Lynn
authors.initials: L
authors.lastname: Wang
authors.forename: Paula
authors.initials: P
authors.lastname: Pittock
authors.forename: Gilles
authors.initials: G
authors.lastname: Lajoie
authors.affiliation: Department of Clinical Neurological Sciences, London Health Sciences Centre, University of Western Ontario , London, Ontario N6A 5A5, Canada.
authors.forename: Shawn Narain
authors.initials: SN
authors.lastname: Whitehead
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004622:Embryo, Mammalian / Q000378:metabolism; D005732:Gangliosides / Q000032:analysis* / Q000302:isolation & purification / Q000494:pharmacology; D018698:Glutamic Acid / Q000633:toxicity*; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D052616:Solid Phase Extraction; D021241:Spectrometry, Mass, Electrospray Ionization; D013110:Sphingosine / Q000737:chemistry
chemical_list: D005732:Gangliosides; D018698:Glutamic Acid; D013110:Sphingosine