Activation of voltage-dependent sodium channels in cultured cerebellar poffule cells induces neurotoxicity that is not mediated by glutamate release.
Exposure of rat cerebellar granule cell cultures to neurotoxins that specifically enhance the open state probability of voltage-dependent Na+ channels, resulted in neuronal death as estimated by a cell viability assay based on fluorescent staining and 51Cr-uptake. Toxicity was detected within 1 h after addition of 100 microM veratridine and was complete within 10-18 h; it was dose-dependent and was found to be completely abolished by tetrodotoxin, an Na+ channel blocker. When veratridine was replaced by an alpha-scorpion toxin, similar observations were done. In contrast, when cultured neurons prepared ffom the cerebral hemisphere of fetal rat brain were exposed to either veratridine or alpha-scorpion toxin for 18 h or even for a longer time of incubation, no neuronal death was observed. DNA fragmentation analysis showed that the toxicity was not mediated by apoptosis. Neuronal death was neither prevented by glutamate receptor antagonists, nor by depletion of endogenous glutamate, nor by voltage sensitive calcium channel antagonists such as omega-Conotoxin-GVIA (N-type channels), omega-Agatoxin-IVA (P-type channels), nimodipine and nitrendipine (L-type channels). Our study indicates that prolonged opening of Na+ channels induced neuronal death of cerebellar granule cells which is not mediated by glutamate and reveals novel neurotoxic mechanism in addition to the well-established excitatory amino acid receptor pathway.
Dargent
B
B
INSERM U374, Institut Jean Roche, Faculté de Medecine Secteur Nord, Marseille, France.
Arsac
C
C
Tricaud
N
N
Couraud
F
F
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001921:Brain / Q000166:cytology; D015220:Calcium Channels / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002860:Chromium Radioisotopes; D004594:Electrophysiology; D018698:Glutamic Acid / Q000378:metabolism*; D015640:Ion Channel Gating / Q000187:drug effects*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D009498:Neurotoxins / Q000037:antagonists & inhibitors / Q000633:toxicity*; D051381:Rats; D017208:Rats, Wistar; D030162:Reptilian Proteins; D012604:Scorpion Venoms / Q000037:antagonists & inhibitors / Q000633:toxicity; D015222:Sodium Channels / Q000187:drug effects* / Q000378:metabolism; D013779:Tetrodotoxin / Q000494:pharmacology; D014701:Veratridine / Q000037:antagonists & inhibitors / Q000633:toxicity
D015220:Calcium Channels; D002860:Chromium Radioisotopes; D009498:Neurotoxins; D030162:Reptilian Proteins; D012604:Scorpion Venoms; D015222:Sodium Channels; C053969:scorpion toxin II, Androctonus; D018698:Glutamic Acid; D013779:Tetrodotoxin; D014701:Veratridine