NeuropharmacologyEcstasy-induced cell death in cortical neuronal cultures is serotonin 2A-receptor-dependent and potentiated under hyperthermia
Section snippets
Materials
Materials for cell cultures were obtained from the following sources: neurobasal medium and supplement B27 from Invitrogen (Paisley, UK); modified Eagle’s medium, phosphate-buffered saline (PBS), HEPES buffer, trypsin/EDTA, penicillin/streptomycin, l-glutamine, collagen-G and poly-l-lysine from Biochrom (Berlin, Germany); multiwell plates from NUNC A/S (Roskilde, Denmark); 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT), 3,3-diaminobenzidine (DAB) and enzyme-standard for
Hyperthermia induced cell death in cortical neuronal cultures
Hyperthermia induced moderate temperature- and time-dependent neurotoxicity at 39 °C and 40 °C, as revealed by LDH measurements at different time-points (24 h, 48 h and 72 h after incubation under hyperthermic conditions) and MTT assay (data not shown). In contrast the temperature of 41 °C proved to be highly toxic already at early time points. This comes in agreement with in vivo experiments which show that hyperthermia in rats with a core temperature above 41 °C can be lethal (Malberg and
Discussion
The key findings of our study are the following: (1) MDMA-induced cortical neurotoxicity in cortical cultures is time- and concentration-dependent, (2) and further potentiated under hyperthermia conditions. (3) MDMA-induced neuronal death follows an apoptotic pattern, (4) which is at least partially mediated by the direct MDMA stimulation of the 5-HT2A-receptor, (5) involves glutamate excitotoxicity, (6) as well as free oxygen radicals. (7) DOI induced a concentration-dependent apoptotic
Conclusion
In conclusion, using a cortical neuronal serum free culture we showed that MDMA neurotoxicity occurs in a dose-, time- and temperature-dependent manner. As the misuse of MDMA as a recreational drug (particularly in crowded and hot environments) can induce hyperthermia, the results presented here corroborate the serious concern previously reported in the literature. This study provides for the first time evidence that direct 5-HT2A-receptor stimulation by MDMA leads to neuronal cortical death.
Acknowledgments
This work was supported by the Hermann & Lilly Schilling Foundation and the Althoff Program of the Charité. J.P.C. was the recipient of a PhD grant from “Fundação para a Ciência e a Tecnologia” (FCT) Portugal (Ref. SFRD/BD/10908/2002). A.R.G. was supported by the “Centro de Matemática da Universidade do Porto” (CMUP), financed by “Fundação para a Ciência e a Tecnologia” (FCT), Portugal, through the programs POCTI and POSI.
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