Gene Therapy for Epilepsies - Gene Delivery and Therapy for Neurological Disorders - page 257

occurrence of high-frequency and multispike complexes and
high-voltage synchronized spike or wave activity in cortical
and hippocampal leads of recording. Also note any synchro-
nous spiking representing interictal activity between seizure
episodes. Note the latency to the first seizure (onset), the total
number of discrete seizure episodes, the total time in seizures
(ictal activity), and the total time in interictal spiking occurring
in the 180 min of recording.

The same procedure described for intrahippocampal KA model can
be easily adapted to the i.c.v. KA model, which induces stronger
and more widespread seizures.
Modifications to the procedure described above are:

3.4.2 Kainic Acid Model:
Intracerebroventricular
Administration

1. Determine the coordinates for lateral ventricle. This is the KA
injection target region. For example, we have used the coordi-
nates (from bregma): AP, −1.0 mm; ML, + 1.5 mm; DV,
−3.7 mm. Prepare the needle according to the determined
depth for injection. For example, using the above DV coordi-
nate, the needle should be 12 mm + 3.7 mm = 15.7 mm.
2. Instead of attaching the bipolar electrode to the stainless can-
nula, implant and secure the hippocampal electrode first, and
then implant the cannula. Since the coordinates for cannula
implantation are close to bregma, implant the cannula last.
3. For i.c.v. KA injection, use a dose of 0.2 μg/3 μl. Dissolve
1 mg of KA in 15 ml saline to give a dilution of 0.2 μg/3 μl.
Aliquot and store at −20 °C until use. Inject 3 μl per animal.

Systemic KA model is possibly the easiest seizure model to employ.
Even if one does not have the setup for EEG monitoring, behav-
ioral seizures can be monitored by simple video recording and sub-
sequent scoring using the Racine motor seizure scale by an
investigator blinded to the treatment group. In the case where
EEG recording is available, simply leave out the cannula implanta-
tion from the above procedure. The KA is administered intraperi-
toneally at 10 mg/kg dose. KA is made up in saline at 1 μg/μl and
injected at 10 μl/g body weight.

3.4.3 Kainic Acid Model:
Systemic Administration

4 Notes

1. All procedures are to be carried out in a biosafety level 2
(BSL2) approved facility and in a Class II tissue culture hood.
2. All materials that have been in contact with AAV should be
decontaminated with Virkon solutions or other suitable
disinfectant.
3. Keep cells in a humidified 37 °C, 5 % CO 2 incubator.

4.1 AAV Vector
Production