title: Embryonic stem cell-derived neuron models of Parkinson's disease exhibit delayed neuronal death.
abstract: Establishment of a Parkinson's disease (PD) neuron model was attempted with mouse embryonic stem (ES) cells. ES cell lines over-expressing mouse nuclear receptor-related 1 (Nurr1), together with human wild-type and alanine 30 --> proline (A30P) and alanine 53 --> threonine (A53T) mutant alpha-synuclein were established and subjected to differentiation into dopaminergic neurons. The ES cell-derived dopaminergic neurons expressing wild-type or mutant alpha-synuclein exhibited the fundamental characteristics consistent with dopaminergic neurons in the substantia nigra. The ES cell-derived PD model neurons exhibited increased susceptibility to oxidative stress, proteasome inhibition, and mitochondrial inhibition. Cell viability of PD model neurons and the control neurons was similar until 28 days after differentiation. Nonetheless, after that time, PD model neurons gradually began to undergo neuronal death over the course of 1 month, showing cytoplasmic aggregate formation and an increase of insoluble alpha-synuclein protein. Such delayed neuronal death was observed in a mutant alpha-synuclein protein level-dependent manner, which was slightly inhibited by a c-jun N-terminal kinase inhibitor and a caspase inhibitor. Such cell death was not observed when the same ES cell lines were differentiated into oligodendrocytes. The ES cell-derived PD model neurons are considered as prospective candidates for a new prototype modelling PD that would allow better investigation of the underlying neurodegenerative pathophysiology.
authors.affiliation: Department of Clinical Neuroscience and Therapeutics, Hiroshima University Graduate School of Biomedical Sciences, Hiroshima, Japan. yamashih@hiroshima-u.ac.jp
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yamashita
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nakamura
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Takahashi
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Nagano
authors.forename: Masanori
authors.initials: M
authors.lastname: Hiji
authors.forename: Takahiro
authors.initials: T
authors.lastname: Hirabayashi
authors.forename: Taku
authors.initials: T
authors.lastname: Amano
authors.forename: Takeshi
authors.initials: T
authors.lastname: Yagi
authors.forename: Norio
authors.initials: N
authors.lastname: Sakai
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Kohriyama
authors.forename: Masayasu
authors.initials: M
authors.lastname: Matsumoto
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D016923:Cell Death / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology / Q000528:radiation effects; D051379:Mice; D008810:Mice, Inbred C57BL; D050505:Mutant Proteins / Q000502:physiology; D016202:N-Methylaspartate / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000473:pathology*; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D018384:Oxidative Stress / Q000502:physiology; D010300:Parkinson Disease / Q000473:pathology* / Q000503:physiopathology; D018408:Patch-Clamp Techniques / Q000379:methods; D013234:Stem Cells / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D050505:Mutant Proteins; C539882:NR4A2 protein, human; D009419:Nerve Tissue Proteins; C539883:Nr4a2 protein, mouse; D057126:Nuclear Receptor Subfamily 4, Group A, Member 2; D014157:Transcription Factors; D051844:alpha-Synuclein; D016202:N-Methylaspartate