title: Direct MRI-guided stereotaxic viral mediated gene transfer of alpha-synuclein in the Göttingen minipig CNS.
abstract: The aim was to establish a non-primate large animal PD model by lentiviral vector mediated mutant alpha-synuclein overexpression in the substantia nigra. Lentivirus encoding A53T alpha-synuclein (6 x 2.5 μl) was stereotaxically injected into the substantia nigra of six adult female Göttingen minipigs. Contralateral control injections encoding enhanced green fluorescent protein (EGFP) were performed. Gait-analysis was performed pre- and postoperatively. PCR of the transgenes and immunohistochemical staining against alpha-synuclein, EGFP, GFAP and TH was performed after 20 weeks. Gait analysis revealed a significant increase in step length and height, and a decrease in the double stand phase. PCR verified the mesencephalic presence of transgenes. IHC analysis showed alpha-synuclein expression in nigral neurons, around the injection tract and in related nigrostriatal projections. The alpha-synuclein positive neurons appeared swollen and vacuolated, in contrast to the EGFP-injected control side. To transduct all nigrostriatal cells with few microinjections, wider dissemination of the transgene must be achieved.
authors.affiliation: Institute of Biomedicine, Faculty of Health Sciences, University of Aarhus, Aarhus, Denmark. angl@ana.au.dk.
authors.forename: Andreas Nørgaard
authors.initials: AN
authors.lastname: Glud
authors.forename: Claus
authors.initials: C
authors.lastname: Hedegaard
authors.forename: Mette Slot
authors.initials: MS
authors.lastname: Nielsen
authors.forename: Jens Christian
authors.initials: JC
authors.lastname: Søorensen
authors.forename: Christian
authors.initials: C
authors.lastname: Bendixen
authors.forename: Poul Henning
authors.initials: PH
authors.lastname: Jensen
authors.forename: Poul Henning
authors.initials: PH
authors.lastname: Mogensen
authors.forename: Knud
authors.initials: K
authors.lastname: Larsen
authors.forename: Carsten Reidies
authors.initials: CR
authors.lastname: Bjarkam
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000008:administration & dosage*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008279:Magnetic Resonance Imaging; D009474:Neurons / Q000378:metabolism; D020734:Parkinsonian Disorders / Q000235:genetics* / Q000378:metabolism; D013378:Substantia Nigra / Q000378:metabolism*; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics* / Q000378:metabolism; D051844:alpha-Synuclein / Q000235:genetics* / Q000378:metabolism
chemical_list: D051844:alpha-Synuclein; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins