title: Direct gene transfer in the Gottingen minipig CNS using stereotaxic lentiviral microinjections.
abstract: We aim to induce direct viral mediated gene transfer in the substantia nigra (SN) of the Gottingen minipig using MRI guided stereotaxic injections of lentiviral vectors encoding enhanced green fluorescent protein (EGFP). Nine female Gottingen minipigs were injected unilaterally into the SN with 6 per 2.5 microliters lentivirus capable of transducing cells and mediating expression of recombinant EGFP. The animals were euthanized after four (n=3) or twenty weeks (n=6). Fresh brain tissue from three animals was used for PCR. The remaining six brains were cryo- or paraffin-sectioned for fluorescence, Nissl-, and immunohistochemical EGFP visualization. EGFP was seen in nigral neurons, axons, glial cells, endothelial cells, and in nigral fibers targeting the striatum. PCR-based detection confirmed the presence of the transgene in SN, whereas all other examined brain areas were negative, indicating that the immunohistochemically detected EGFP in the striatum derived from transfected nigral cells.
authors.affiliation: Institute of Anatomy, Faculty of Health Sciences, University of Aarhus, DK-8000 Aarhus C, Denmark. angl@ana.au.dk.
authors.forename: Andreas
authors.initials: A
authors.lastname: Norgaard 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: Sørensen
authors.forename: Christian
authors.initials: C
authors.lastname: Bendixen
authors.forename: Poul Henning
authors.initials: PH
authors.lastname: Jensen
authors.forename: Knud
authors.initials: K
authors.lastname: Larsen
authors.forename: Carsten Reidies
authors.initials: CR
authors.lastname: Bjarkam
mesh_terms: D000818:Animals; D001522:Behavior, Animal / Q000502:physiology; D001921:Brain / Q000821:virology; D004247:DNA / Q000008:administration & dosage / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics* / Q000254:growth & development; D008845:Microinjections; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013238:Stereotaxic Techniques; D013552:Swine; D013556:Swine, Miniature
chemical_list: C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA