title: DNA damage and ubiquitinated neuronal inclusions in the substantia nigra and striatum of mice following MDMA (ecstasy).
abstract: BACKGROUND
  3,4-Methylenedioxymethamphetamine (MDMA) is an amphetamine derivative, which is neurotoxic to both serotonin (5HT) and dopamine (DA) nerve terminals. Previous reports, carried out in rodents and non-human primates, demonstrated neurotoxicity to monoamine axon terminals, although no study has analyzed nigral and striatal cell bodies at the sub-cellular level.
  
  
  OBJECTIVE
  In this study, we examined intrinsic nigral and striatal cells, and PC12 cell cultures to evaluate whether, in mice, MDMA might affect nigral and striatal cell bodies.
  
  
  METHODS
  After administering MDMA, we analyzed effects induced in vivo and in vitro using high-performance liquid chromatography (HPLC) analysis, light- and electron microscopy with immunocytochemistry, and DNA comet assay.
  
  
  RESULTS
  We found that MDMA (5 mg/kg x4, 2 h apart), besides a decrease of nigrostriatal DA innervation and 5HT loss, produces neuronal inclusions within nigral and intrinsic striatal neurons consisting of multi-layer ubiquitin-positive whorls extending to the nucleus of the cell. These fine morphological changes are associated with clustering of heat shock protein (HSP)-70 in the nucleus, very close to chromatin filaments. In the same experimental conditions, we could detect oxidation of DNA bases followed by DNA damage. The nature of inclusions was further investigated using PC12 cell cultures.
  
  
  CONCLUSIONS
  The present findings lead to re-consideration of the neurotoxic consequences of MDMA administration. In fact, occurrence of ubiquitin-positive neuronal inclusions and DNA damage both in nigral and striatal cells sheds new light into the fine alterations induced by MDMA, also suggesting the involvement of nuclear and cytoplasmic components of the ubiquitin-proteasome pathway in MDMA toxicity.
authors.affiliation: Department of Human Morphology and Applied Biology, University of Pisa, 56126 Pisa, Italy. f.fornai@med.unipi.it
authors.forename: F
authors.initials: F
authors.lastname: Fornai
authors.forename: P
authors.initials: P
authors.lastname: Lenzi
authors.forename: G
authors.initials: G
authors.lastname: Frenzilli
authors.forename: M
authors.initials: M
authors.lastname: Gesi
authors.forename: M
authors.initials: M
authors.lastname: Ferrucci
authors.forename: G
authors.initials: G
authors.lastname: Lazzeri
authors.forename: F
authors.initials: F
authors.lastname: Biagioni
authors.forename: M
authors.initials: M
authors.lastname: Nigro
authors.forename: A
authors.initials: A
authors.lastname: Falleni
authors.forename: M
authors.initials: M
authors.lastname: Giusiani
authors.forename: A
authors.initials: A
authors.lastname: Pellegrini
authors.forename: F
authors.initials: F
authors.lastname: Blandini
authors.forename: S
authors.initials: S
authors.lastname: Ruggieri
authors.forename: A
authors.initials: A
authors.lastname: Paparelli
mesh_terms: D000818:Animals; D003342:Corpus Striatum / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D004249:DNA Damage; D004298:Dopamine / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000633:toxicity*; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D016716:PC12 Cells; D051381:Rats; D018490:Serotonin Agents / Q000633:toxicity*; D013378:Substantia Nigra / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D025801:Ubiquitin / Q000378:metabolism*
chemical_list: D018490:Serotonin Agents; D025801:Ubiquitin; D018817:N-Methyl-3,4-methylenedioxyamphetamine; D004298:Dopamine