Monosialoganglioside GM1 protects against anoxia-induced neuronal death in vitro.
Excitatory dicarboxylic amino acid neurotransmitters, particularly glutamate, have been implicated in mediating neuronal cell injury in brain ischemia-anoxia, epilepsy, and stroke. Glutamate neurotoxicity has been demonstrated in several in vitro models, as well as its prevention by a variety of agents, including several sialic acid-containing glycosphingolipid species, gangliosides. We have now examined ganglioside effects in anoxic exposed cultures of granule cells from Postnatal Day 8 rat cerebellum. Cells between 10 and 12 days in vitro were placed into an anoxic atmosphere or subjected to a chemical model of anoxia by a pulse exposure to rotenone. Widespread neuronal degeneration of neuronal cell bodies and their associated neurite network was seen the following day. These effects on cell vitality at the morphological level were quantitatively confirmed by measuring the photometric reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide to a blue formazan product. This neuronal injury was abolished by the specific N-methyl-D-aspartate receptor noncompetitive antagonists Mg2+, phencyclidine and MK-801, suggesting that this subtype of glutamate receptor is involved in the pathogenesis of anoxic granule cell injury. Pretreatment for 30 to 60 min or more or concurrent treatment with ganglioside GM1 largely prevented the ensuing neuronal death (ED50 = 25 microM), even 4 days later. Degeneration induced by exogenous glutamate was equally reduced. Asialo GM1 (lacking sialic acid) was ineffective. These results are consistent with the observed beneficial effects of the gangliosides in ischemic brain injury models in vivo.
Skaper
S D
SD
Fidia Research Laboratories, Department of CNS Research, Abano Terme, Italy.
Facci
L
L
Milani
D
D
Leon
A
A
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D005677:G(M1) Ganglioside / Q000494:pharmacology*; D005732:Gangliosides / Q000494:pharmacology*; D000860:Hypoxia / Q000139:chemically induced / Q000473:pathology*; D008858:Microscopy, Phase-Contrast; D009474:Neurons / Q000473:pathology / Q000502:physiology*; D009584:Nitrogen; D012402:Rotenone; D013778:Tetrazolium Salts; D013844:Thiazoles
D004396:Coloring Agents; D005732:Gangliosides; D013778:Tetrazolium Salts; D013844:Thiazoles; C025447:sialogangliosides; D012402:Rotenone; D005677:G(M1) Ganglioside; C022616:thiazolyl blue; D009584:Nitrogen