PMID- 28745630
OWN - NLM
STAT- MEDLINE
DCOM- 20180124
LR  - 20190713
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 125
DP  - 2017 Jul 13
TI  - Subpial Adeno-associated Virus 9 (AAV9) Vector Delivery in Adult Mice.
LID - 10.3791/55770 [doi]
LID - 55770
AB  - 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 microL; 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.
FAU - Tadokoro, Takahiro
AU  - Tadokoro T
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego.
FAU - Miyanohara, Atsushi
AU  - Miyanohara A
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego.
FAU - Navarro, Michael
AU  - Navarro M
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego.
FAU - Kamizato, Kota
AU  - Kamizato K
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego.
FAU - Juhas, Stefan
AU  - Juhas S
AD  - Institute of Animal Physiology and Genetics, Czech Academy of Sciences.
FAU - Juhasova, Jana
AU  - Juhasova J
AD  - Institute of Animal Physiology and Genetics, Czech Academy of Sciences.
FAU - Marsala, Silvia
AU  - Marsala S
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego.
FAU - Platoshyn, Oleksandr
AU  - Platoshyn O
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego.
FAU - Curtis, Erik
AU  - Curtis E
AD  - Department of Neurosurgery, University of California, San Diego.
FAU - Gabel, Brandon
AU  - Gabel B
AD  - Department of Neurosurgery, University of California, San Diego.
FAU - Ciacci, Joseph
AU  - Ciacci J
AD  - Department of Neurosurgery, University of California, San Diego.
FAU - Lukacova, Nada
AU  - Lukacova N
AD  - Institute of Neurobiology, Slovak Academy of Sciences.
FAU - Bimbova, Katarina
AU  - Bimbova K
AD  - Institute of Neurobiology, Slovak Academy of Sciences.
FAU - Marsala, Martin
AU  - Marsala M
AD  - Neuroregeneration Laboratory, Department of Anesthesiology, University of 
      California, San Diego; Institute of Neurobiology, Slovak Academy of Sciences; 
      mmarsala@ucsd.edu.
LA  - eng
GR  - P30 AI036214/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20170713
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Animals
MH  - Brain/metabolism
MH  - Dependovirus/*genetics
MH  - Female
MH  - Genetic Vectors/genetics/*metabolism
MH  - Green Fluorescent Proteins/genetics
MH  - Male
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Microscopy, Fluorescence
MH  - Spinal Cord/metabolism
MH  - Video Recording
PMC - PMC5612300
EDAT- 2017/07/27 06:00
MHDA- 2018/01/25 06:00
PMCR- 2019/07/13
CRDT- 2017/07/27 06:00
PHST- 2017/07/27 06:00 [entrez]
PHST- 2017/07/27 06:00 [pubmed]
PHST- 2018/01/25 06:00 [medline]
PHST- 2019/07/13 00:00 [pmc-release]
AID - 55770 [pii]
AID - 10.3791/55770 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Jul 13;(125):55770. doi: 10.3791/55770.