Biology Reference
In-Depth Information
5. Scoring of immobility begins after 2 min have elapsed. Scoring
can be performed after the experiment has been completed
using the recorded video, or one mouse can be scored while
the animals are being tested leaving one to be scored at a later
time from the recorded video. Only one mouse can be scored
at a time. Each time the mouse ceases struggling and swim-
ming movements and appears to be resting or floating (making
only the minimal movements needed to stay afloat), the stop-
watch is started. The watch is stopped when the bout of immo-
bility ends. The watch is similarly started and stopped each
time the mouse enters into and then ends a bout of immobility.
At the end of the session the cumulative time spent in immo-
bility is recorded off the stopwatch. Scoring continues for the
remainder of the test period, which is predetermined and is
typically from 4 to 8 additional minutes.
6. Mice are removed from the water at the end of the test period
and placed individually into two clean mouse cages lined with
several dry paper or cloth towels. They are dried with the tow-
els and then remain in the drying cages for 5-10 min before
being returned to their homecages.
7. The beakers are emptied and refilled with clean water for the
next pair of mice. The process is repeated for remaining mice.
8. Cumulative immobility over the test period is reported.
Individually house mice for at least 5 days in usual mouse cages
prior to starting the habituation (see Note 4).
3.3 Mouse
Hyponeophagia
Day 1
3.3.1 Habituation
(Days 1-3)
1. Dilute Carnation sweetened condensed milk 1:3 in water and
mix well.
2. Partially fill clean water bottles with the diluted milk solution.
Use at room temperature.
3. A bottle containing the milk solution is put on each cage in the
place of the normal water for a 20-min period.
4. Milk bottles are then removed and normal water bottles are
returned to all cages.
5. The bottles of milk can be reused for all habituation days (for
the same animal) if they are refrigerated after each session and
brought to room temperature before use. Discard after 3 days
of use.
Days 2-3
1. EPO (500 U/kg) is injected intraperitoneally (i.p.) and vehicle
is similarly administered to a second group of mice (8/
group).
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