Gene Delivery and Gene Therapy for Alzheimer’s Disease - Gene Delivery and Therapy for Neurological Disorders - page 109

Fig. 2 Stereotaxic injection of AAV 7ND and AAV-GFP into the cerebral cortex and hippocampus. ( a ) Mice at 3
months of age were stereotactically injected with AAV-7ND or AAV-GFP into the cerebral cortex or hippocam-
pus at different doses [3 × 10 8 , 1.2 × 10 9 , 5 × 10 9 , and 2 × 10 10 viral particles (VP)/brain] and sacrifi ced 1 month
postinjection. ( b , c ) Sustained expression of 7ND/CCL2 was observed after cortical ( b ) hippocampal ( c ) injec-
tion of AAV-7ND or AAV-GFP (2 × 10 10 VP/2 µl/site). Cited from [ 85 ]

3.3 Delivery of Virus
to Mouse Brain In Vivo

All procedures should be approved by Institutional Animal Care and
Use Committee

1. Determine stereotaxic coordinates according to the brain area
of interest. To deliver AAV to the hippocampus, we use a site
in the dorsal hippocampus in the apical dendritic zones of the
CA1 region near the hippocampal fi ssure ( see Fig. 2a , a scheme
showing injection sites in the hippocampus and cortex of con-
trol AAV-GFP or AAV-7ND). For delivery to this site, the
skull should be marked bilaterally ( see step 3 of Sect. 3.3.3 ) at
the following stereotaxic coordinates of bregma: anterioposte-
rior (AP) = −1.8 mm, mediolateral (ML) = −/+1.8 mm, dorso-
ventral (DV) = −1.8 mm.

3.3.1 Stereotaxic
Coordinates

All surgeries should be performed in a dedicated surgery room,
using sterile instruments, surgical gloves, and aseptic procedures.
To prepare the animal for injection:

3.3.2 Anesthesia
and Brain Microinjections

Aseptic Procedures
and Anesthesia

1. Trim the fur with scissors, and then shave the surgical site (top
of the head for brain microinjection) using a small razor.
Disinfect the site with betadine solution followed by alcohol.
Repeat the betadine/alcohol treatment three times.
2. Perform all surgical brain microinjections under general
anesthesia induced either by a 5 % isofl urane/O 2 mixture
using a research anesthesia machine connected to a chamber
and scavenger circuits or by IP injection with 65 mg/kg
ketamine, 13 mg/kg xylazine, and 2 mg/kg acepromazine
prior to the intracerebral injection. After induction of anes-
thesia, maintain animals at a proper level of anesthesia,
which can be determined by absence of the corneal refl ex,
with 1-3 % isofl urane.