Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice

0483870 - ÚŽFG 2018 RIV US eng J - Článek v odborném periodiku
Tadokoro, T. - Miyanohara, A. - Navarro, M. - Kamizato, K. - Juhás, Štefan - Juhásová, Jana - Maršala, S. - Platoshyn, O. - Curtis, E. - Gabel, B. - Ciacci, J. D. - Lukáčová, N. - Bimbová, K. - Maršala, M.
Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice.
Jove-Journal of Visualized Experiments. Roč. 125, č. 13 (2017), č. článku e55770. ISSN 1940-087X
Grant CEP: GA MŠMT(CZ) LO1609
Institucionální podpora: RVO:67985904
Klíčová slova: AAV9 * adult mouse
Obor OECD: Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction)
Impakt faktor: 1.184, rok: 2017

The successful development of a subpial adeno-associated virus 9 (AAV9) vector delivery technique in adult rats and pigs has been reported on previously. Using subpially-placed polyethylene catheters (PE-10 or PE-5) for AAV9 delivery, potent transgene expression through the spinal parenchyma (white and gray matter) in subpially-injected spinal segments has been demonstrated. Because of the wide range of transgenic mouse models of neurodegenerative diseases, there is a strong desire for the development of a potent central nervous system (CNS)-targeted vector delivery technique in adult mice. Accordingly, the present study describes the development of a spinal subpial vector delivery device and technique to permit safe and effective spinal AAV9 delivery in adult C57BL/6J mice. In spinally immobilized and anesthetized mice, the pia mater (cervical 1 and lumbar 1-2 spinal segmental level) was incised with a sharp 34 G needle using an XYZ manipulator. A second XYZ manipulator was then used to advance a blunt 36G needle into the lumbar and/or cervical subpial space. The AAV9 vector (3-5 mu L, 1.2 x 10(13) genome copies (gc)) encoding green fluorescent protein (GFP) was then injected subpially. After injections, neurological function (motor and sensory) was assessed periodically, and animals were perfusion-fixed 14 days after AAV9 delivery with 4% paraformaldehyde. Analysis of horizontal or transverse spinal cord sections showed transgene expression throughout the entire spinal cord, in both gray and white matter. In addition, intense retrogradely-mediated GFP expression was seen in the descending motor axons and neurons in the motor cortex, nucleus ruber, and formatio reticularis. No neurological dysfunction was noted in any animals. These data show that the subpial vector delivery technique can successfully be used in adult mice, without causing procedure-related spinal cord injury, and is associated with highly potent transgene expression throughout the spinal neuraxis.
Trvalý link: http://hdl.handle.net/11104/0279041