Biology Reference
In-Depth Information
1. EPO and vehicle are administered as on the prior 3 days to the
same groups of rats. This is the fourth consecutive day of treat-
ment. Time injections so that pairs of rats are injected every
15-20 min. Swim testing starts for the first pair of rats 4 h after
they are injected.
2. The set-up is identical to that on day 1 (see Note 6). The two
cylinders are placed on a cart or table at one end of the room,
with a plain white surface (such as a bare wall) behind them
and a white divider between them. Visual stimuli should be the
same on the outer sides of both cylinders. The camera is set on
a tripod at the other end of the room where the experimenter
also sits (equidistant from both cylinders). Ensure that normal
overhead lighting falls evenly on both cylinders.
3. All rats to be tested are brought in their homecages into a
quiet holding room near the testing room and rats acclimate
for 1 h before the first pair is tested.
4. Attach hose or tubing to faucet and fill cylinders while adjust-
ing the hot and cold flows so that the temperature of water is
23-25 °C. Fill to a depth of 45 cm.
5. Two rats at a time are brought from the holding room into
the testing room and gently lowered into the two cylinders.
Each rat should be tested in the same cylinder as the prior day.
The experiment timer is started and runs for 10 min.
6. Scoring of immobility can be performed after the experiment
has been completed using the video recording, or one rat can
be scored while the animals are being tested leaving one to be
scored at a later time from the recorded video. Only one animal
can be scored at a time. Sessions are videotaped from the side
and scored by using a behavioral sampling technique to assess
the primary behavior for each 5-s interval. Three behaviors are
scored: (a) climbing, rat is thrashing in an upward motion
along the sides of the tank; (b) swimming, movement across
the tank into another quadrant; and (c) immobility, when the
animal made the minimal amount of movement required to
stay afloat.
7. At the end of the 10 min session, the rats are removed from the
water and placed in clean rat cages lined with several dry paper
or cloth towels. They are dried with the towels and then remain
in the drying cages for 10 min before being returned to their
homecages.
8. Cylinders are emptied and refilled with clean water for the next
pair of rats. The process is repeated for remaining animals.
9. Scored behaviors are quantified and reported for the entire
test period or as they occur over defined intervals of the test
(see Fig.
1
).
3.1.2
Swim Day 2
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