Gene Delivery and Gene Therapy for Alzheimer’s Disease - Gene Delivery and Therapy for Neurological Disorders - page 112

3. On the walls of the tank, place spatial cues for animal
orientation, such as pictures or shapes.
4. At the end of one arm, submerge a 10 cm circular Plexiglass
platform 1 cm below the arm, in order to hide it from the
animal.
5. Record animal behavior using a tracking system, such as
Ethovision System 3.1 (Noldus Information Technology
Inc., Leesburg, MA, USA) and a CCD camera suspended
170 cm above the liquid surface ( see Fig. 3a for sample of
recorded trial).

1. Trial numbers and the organization of testing may be con-
structed according to the needs of your study. Here, we give an
explanation of our RAWM testing method, which is based on
the “win-stay” RAWM protocol for mice developed by
Arendash et al. [ 106 ].
2. Organization: On day 1 of testing, 15 trials (12 trials with vis-
ible platform, followed by 3 trials with hidden platform) are
run in fi ve blocks of three. Run a cohort of four mice sequen-
tially for each block. After each three-trial block, run a second
cohort of mice in order to create an extended rest period of
the fi rst cohort before exposure to the second block. Keep
the target arm location constant for a given mouse through-
out the test.
3. Timing and scoring: Each trial lasts one minute, and an error
is scored each time the body of the mouse, excluding the tail,
enters the wrong arm, enters the arm with the platform but
does not climb on it, or does not make a choice for 20 s
( see Fig. 3b for sample results, showing signifi cantly fewer
errors on day 2-10 in mice injected with AAV-IL10 as com-
pared to control mice). Each trial ends when the mouse climbs
onto and remains on the hidden platform for 10 s. The mouse
is given 20 s to rest on the platform between each trial.
4. On day 2, run the animals in exactly the same manner as day 1,
except that the platform is hidden for all trials. Swimming
speed (measured by automatic tracking system) may also be
compared between animals ( see Fig. 3c for sample results).
5. Testing of short-term memory recall: to test the short-term
memory recall, give each animal an additional break of
30 min between blocks 4 and 5 on day 1 and blocks 9 and
10 on day 2.
6. The errors on each block can be averaged and used for statisti-
cal analysis.

3.5.2 Testing Animals
in the Maze