title: Reconstruction and quantitative evaluation of dopaminergic innervation of striatal neurons in dissociated primary cultures.
abstract: Repairing the nigrostriatal pathway is expected to become a future treatment strategy for Parkinson disease. Our aim is to establish an in vitro model for the quantitative analysis of the nigrostriatal projections of dopaminergic neurons using primary dissociated neruons. To form the mesencephalic cell region, mesencephalic cells derived from rat embryos were plated within an isolation wall, which was removed after cell adhesion to the coverslip. After incubation for 11 days, the dopaminergic neurites extending to the outside of the mesencephalic cell region were mainly axons. Treatment with glial cell line-derived neurotrophic factor for 11 days significantly promoted the outgrowth of dopaminergic axons from the mesencephalic cell region in a concentration-dependent manner. When striatal cells were plated outside the mesencephalic cell region, dopaminergic neurites were remarkably extended to the striatal cell region. Moreover, immunocytochemistry for tyrosine hydroxylase and synaptophysin revealed that dopaminergic axons formed synapses with striatal neurons. By contrast, spinal cells did not increase dopaminergic neurite outgrowth. These results indicate that the present method is valuable for evaluating nigrostriatal projections in vitro.
authors.affiliation: Department of Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, 46-29 Yoshida-shimoadachi-cho, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Seiko
authors.initials: S
authors.lastname: Wakita
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Izumi
authors.forename: Takaaki
authors.initials: T
authors.lastname: Matsuo
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Kume
authors.forename: Yuki
authors.initials: Y
authors.lastname: Takada-Takatori
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Sawada
authors.forename: Akinori
authors.initials: A
authors.lastname: Akaike
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D003342:Corpus Striatum / Q000166:cytology*; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004298:Dopamine / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004622:Embryo, Mammalian; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels / Q000378:metabolism; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000494:pharmacology; D006801:Humans; D008636:Mesencephalon / Q000166:cytology; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009415:Nerve Net / Q000502:physiology*; D016501:Neurites / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000737:chemistry / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D013116:Spinal Cord / Q000166:cytology; D013569:Synapses / Q000187:drug effects / Q000502:physiology; D016708:Synaptophysin / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D051676:G Protein-Coupled Inwardly-Rectifying Potassium Channels; D051100:Glial Cell Line-Derived Neurotrophic Factor; C489787:MAP2 protein, rat; D008869:Microtubule-Associated Proteins; D016708:Synaptophysin; D014446:Tyrosine 3-Monooxygenase; D004298:Dopamine