Increased synthesis of specific proteins during glutamate-induced neuronal death in cerebellar culture.
We have previously shown that glutamate-induced neurotoxicity is mediated by a sodium-chloride component and a calcium component in our cerebellar granule cell culture. In order to further characterize these two different components, the time course of neuronal death induced by glutamate (100 microM) in basal solution and in low sodium-chloride solution was studied by morphological and biochemical criteria. As shown by phase-contrast microscopy, cerebellar granule cells exhibited clear neuronal degeneration within 4 h after exposure to this excitotoxin. These morphological changes correlated [35S]methionine incorporation into proteins which rapidly declined during the first hour of treatment. Qualitative change in [35S]methionine incorporation into proteins was further investigated by two-dimensional gel electrophoresis performed after glutamate exposure in basal solution and in low sodium-chloride solution. Most of the proteins showed a decreased labelling after glutamate exposure as expected, but some polypeptides showed an increased labelling or appeared to be newly synthesized. Furthermore, a different pattern of protein synthesis was observed when glutamate exposure was performed in basal solution or in low sodium-chloride solution. The identification of these polypeptides and their implication in the neuronal death are discussed.
Dessi
F
F
INSERM U29, Paris, France.
Ben-Ari
Y
Y
Charriaut-Marlangue
C
C
D000818:Animals; D015153:Blotting, Western; D016923:Cell Death / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D018698:Glutamic Acid / Q000494:pharmacology*; D008715:Methionine / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D009474:Neurons / Q000187:drug effects*; D051381:Rats; D013462:Sulfur Radioisotopes
D009419:Nerve Tissue Proteins; D013462:Sulfur Radioisotopes; D018698:Glutamic Acid; D008715:Methionine