title: GM1 ganglioside reduces glutamate toxicity to cortical cells. Lowered LDH release and preserved membrane integrity.
abstract: As an in vitro model of CNS excitatory amino acid (EAA) injury, rat cortical neuronal cultures were challenged with glutamate (0.5 or 10 mM) and the levels of released lactate dehydrogenase (LDH) were monitored at 1 h, 1, 2, and 7 d. LDH release is correlated with levels of plasma membrane damage. GM1 has been shown to be continuously distributed on the outer surface of CNS cellular membranes. By staining for the distribution of endogenous GM1 ganglioside using cholera toxin/antitoxin immunohistochemistry, we were able to assess morphologically cellular plasma membrane integrity after damage. We used these two measures (LDH and GM1 localization) to study the neuroprotective effects of exogenous GM1 ganglioside to further elucidate its mechanism. Cortical cultures derived from 15-d rat fetuses were subjected to the glutamate challenge for 30 min. Parallel cultures were either pre- or post-treated with 80 microM of GM1. Exposure to 10 mM glutamate caused a highly significant increase in LDH release at 1-48 h. Pretreatment with GM1 reduced the release, whereas posttreatment reduced the LDH release even more. Plasma membrane changes observed by the GM1 immunohistochemistry reflected the LDH release data. All cultures treated with GM1 evidenced substantial structural integrity (continuous staining of GM1 along perikarya and processes) as compared to untreated cultures. These data support our hypothesis that GM1 treatment (pre- and post-) reduces plasma membrane damage.
authors.affiliation: Department of Psychiatry, Columbia University, College of Physicians and Surgeons, New York, NY 10032.
authors.forename: H
authors.initials: H
authors.lastname: Laev
authors.forename: S P
authors.initials: SP
authors.lastname: Mahadik
authors.forename: J L
authors.initials: JL
authors.lastname: Bonheur
authors.forename: N
authors.initials: N
authors.lastname: Hernandez
authors.forename: S E
authors.initials: SE
authors.lastname: Karpiak
mesh_terms: D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D002772:Cholera Toxin / Q000276:immunology; D018691:Excitatory Amino Acid Antagonists; D005677:G(M1) Ganglioside / Q000494:pharmacology*; D005971:Glutamates / Q000633:toxicity*; D018698:Glutamic Acid; D007150:Immunohistochemistry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D010088:Oxidoreductases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates; D005677:G(M1) Ganglioside; D018698:Glutamic Acid; D002772:Cholera Toxin; D010088:Oxidoreductases; D007770:L-Lactate Dehydrogenase