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Fig. 1
Viral infusion into mice. (
a
,
b
) Apparatus used for viral infusion. Stereotactic frame with a syringe pump
holding Hamilton syringe connected to infusion pipette via polyethylene tubing. (
c
,
d
) Images of a mouse
mounted on the stereotactic frame with infusion pipette inserted into the brain through one of the two holes
drilled in the skull. (
e
) Close-up of the exposed skull before drilling with Bregma and Lambda indicated
are longer than that of the recombinant HSV particles, the time of
infection between rounds (18-24 h) has been optimized to obtain
high yields of recombinant virus while minimizing the production
of helper virus. It is important, therefore, not to exceed these times
when performing this procedure.
In this procedure, animals are anesthetized, holes drilled in the
skull, and small volumes of recombinant virus are slowly infused
into the LA using glass pipettes (normally used for patch clamping
of neurons) (Fig.
1
). After a suitable time for recovery, animals are
then placed back into their home cage for 3-4 days to allow trans-
gene expression before behavioral experiments are performed.
3.2 Infusion of
Amplicons into the LA
In Vivo
1. Weigh and suitably anesthetize animal. Ensure an adequate
level of anesthesia has been induced before proceeding with
surgery.
3.2.1 Preparation
of Animal
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