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Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice
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SYSNO ASEP 0483870 Druh ASEP J - Článek v odborném periodiku Zařazení RIV J - Článek v odborném periodiku Poddruh J Článek ve WOS Název Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice Tvůrce(i) Tadokoro, T. (US)
Miyanohara, A. (US)
Navarro, M. (US)
Kamizato, K. (US)
Juhás, Štefan (UZFG-Y) RID, ORCID
Juhásová, Jana (UZFG-Y) RID, ORCID
Maršala, S. (US)
Platoshyn, O. (US)
Curtis, E. (US)
Gabel, B. (US)
Ciacci, J. D. (US)
Lukáčová, N. (SK)
Bimbová, K. (SK)
Maršala, M. (US)Číslo článku e55770 Zdroj.dok. Jove-Journal of Visualized Experiments - ISSN 1940-087X
Roč. 125, č. 13 (2017)Poč.str. 10 s. Forma vydání Online - E Jazyk dok. eng - angličtina Země vyd. US - Spojené státy americké Klíč. slova AAV9 ; adult mouse Vědní obor RIV EI - Biotechnologie a bionika Obor OECD Technologies involving the manipulation of cells, tissues, organs or the whole organism (assisted reproduction) CEP LO1609 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy Institucionální podpora UZFG-Y - RVO:67985904 UT WOS 000407455900037 EID SCOPUS 85024404853 DOI 10.3791/55770 Anotace 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. Pracoviště Ústav živočišné fyziologie a genetiky Kontakt Jana Zásmětová, knihovna@iapg.cas.cz, Tel.: 315 639 554 Rok sběru 2018
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