title: The neurotoxicity of hallucinogenic amphetamines in primary cultures of hippocampal neurons.
abstract: 3,4-Methylenedioxymethamphetamine (MDMA or "Ecstasy") and 2,5-dimethoxy-4-iodoamphetamine hydrochloride (DOI) are hallucinogenic amphetamines with addictive properties. The hippocampus is involved in learning and memory and seems particularly vulnerable to amphetamine's neurotoxicity. We evaluated the neurotoxicity of DOI and MDMA in primary neuronal cultures of hippocampus obtained from Wistar rat embryos (E-17 to E-19). Mature neurons after 10 days in culture were exposed for 24 or 48 h either to MDMA (100-800 μM) or DOI (10-100 μM). Both the lactate dehydrogenase (LDH) release and the tetrazolium-based (MTT) assays revealed a concentration- and time-dependent neuronal death and mitochondrial dysfunction after exposure to both drugs. Both drugs promoted a significant increase in caspase-8 and caspase-3 activities. At concentrations that produced similar levels of neuronal death, DOI promoted a higher increase in the activity of both caspases than MDMA. In the mitochondrial fraction of neurons exposed 24h to DOI or MDMA, we found a significant increase in the 67 kDa band of apoptosis inducing factor (AIF) by Western blot. Moreover, 24h exposure to DOI promoted an increase in cytochrome c in the cytoplasmatic fraction of neurons. Pre-treatment with an antibody raised against the 5-HT(2A)-receptor (an irreversible antagonist) greatly attenuated neuronal death promoted by 48 h exposure to DOI or MDMA. In conclusion, hallucinogenic amphetamines promoted programmed neuronal death involving both the mitochondria machinery and the extrinsic cell death key regulators. Death was dependent, at least in part, on the stimulation of the 5-HT(2A)-receptors.
authors.affiliation: REQUIMTE (Rede de Química e Tecnologia), Laboratório de Toxicologia, Departamento de Ciências Biológicas, Faculdade de Farmácia, Universidade do Porto, Rua de Jorge Viterbo Ferreira, 228, 4050-313 Porto, Portugal. joaocapela@ff.up.pt
authors.forename: João Paulo
authors.initials: JP
authors.lastname: Capela
authors.forename: Silvana
authors.initials: S
authors.lastname: da Costa Araújo
authors.forename: Vera Marisa
authors.initials: VM
authors.lastname: Costa
authors.forename: Karsten
authors.initials: K
authors.lastname: Ruscher
authors.forename: Eduarda
authors.initials: E
authors.lastname: Fernandes
authors.forename: Maria de Lourdes
authors.initials: Mde L
authors.lastname: Bastos
authors.forename: Ulrich
authors.initials: U
authors.lastname: Dirnagl
authors.forename: Andreas
authors.initials: A
authors.lastname: Meisel
authors.forename: Félix
authors.initials: F
authors.lastname: Carvalho
mesh_terms: D000662:Amphetamines / Q000633:toxicity*; D000818:Animals; D051033:Apoptosis Inducing Factor / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D045304:Cytochromes c / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005865:Gestational Age; D006213:Hallucinogens / Q000633:toxicity*; D006624:Hippocampus / Q000187:drug effects* / Q000196:embryology / Q000378:metabolism / Q000473:pathology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000633:toxicity*; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D020258:Neurotoxicity Syndromes / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D061251:Primary Cell Culture; D051381:Rats; D017208:Rats, Wistar; D044402:Receptor, Serotonin, 5-HT2A / Q000187:drug effects / Q000378:metabolism; D058830:Serotonin 5-HT2 Receptor Antagonists / Q000494:pharmacology; D013997:Time Factors
chemical_list: C494831:Aifm1 protein, rat; D000662:Amphetamines; D051033:Apoptosis Inducing Factor; D015415:Biomarkers; D006213:Hallucinogens; D044402:Receptor, Serotonin, 5-HT2A; D058830:Serotonin 5-HT2 Receptor Antagonists; D045304:Cytochromes c; D007770:L-Lactate Dehydrogenase; C505410:Casp3 protein, rat; C505528:Casp8 protein, rat; D053148:Caspase 3; D053181:Caspase 8; D018817:N-Methyl-3,4-methylenedioxyamphetamine; C015952:4-iodo-2,5-dimethoxyphenylisopropylamine