Direct gene transfer in the Goettingen minipig CNS using stereotaxic lentiviral microinjections

• Published in: Acta neurobiologiae experimentalis Vol. 70; no. 3; pp. 308 - 315
• Main Authors: Glud, AN, Hedegaard, C, Nielsen, MS, Soerensen, J C, Bendixen, C, Jensen, PH, Larsen, K, Bjarkam, C R
• Format: Journal Article
• Language: English
• Published: 01.01.2010
• Subjects: Lentivirus
• ISSN: 0065-1400

We aim to induce direct viral mediated gene transfer in the substantia nigra (SN) of the Goettingen minipig using MRI guided stereotaxic injections of lentiviral vectors encoding enhanced green fluorescent protein (EGFP). Nine female Goettingen minipigs were injected unilaterally into the SN with 6x2.5 mu l 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.