Effect of Ethanol on Anxious Behaviors in Rats

Lauren Brasseur, Cayleigh Celli, Mykelle Morrison

Department of Psychology, University of Washington
Seattle, Washington 98105

Introduction
Background Info
Mice that were exposed to stressful stimuli before entering an open field had a very low
center to peripheral activity ratio in the first 6 minutes relative to the rest of their time in
the open field. In comparison, control mice had a fairly constant center to peripheral
activity ratio during their entire open field exposure.2 Based on these conclusions we
can infer that a stressful stimulus like alcohol will decrease time spent in the center of
the open field of ethanol rats between the first and second tests, but the time spent in the
center of the open field between the first and second tests should remain the same for
control rats.

Results
For our first two tests we calculated the delta between tests one and two of each
subject. Using the absolute values of these numbers we ran independent t tests (alpha
level of .05 for all tests) between the control and ethanol subject groups.
Change in Time Spent in Center of Open Field Between Tests

Rats were exposed to either water or ethanol and assessed for locomotor activity.
During the first five minute interval ethanol consuming rats made approximately 135%
more square crossings than the water consuming rats.1 Based on this study, we can
conclude that upon exposure to alcohol, the ethanol rats will perform more line
crossings in their second open field test than their first, in comparison to the rats
receiving no ethanol.

Control
Linear (Control)
Ethanol
Linear (Ethanol )

When mice are introduced to a novel environment it is common for animals to display
increased grooming activity. It has also been observed that in rats, minor stressors, like
handling, transportation, and novelty lead to increased grooming behaviors.3 Based on
this study, we can conclude that the alcohol group, having been exposed to more
stressors, will groom more than the control group in their second test.
Based on our research, we have determined that amount of grooming, number of line
crossings, and time spent in the center of the open field are all indicators of anxiety in
rats. The objective of our study is to explain how ethanol can alter these behaviors.
Research Question:
How does ethanol ingestion affect anxious behaviors in rats when exposed to an
unfamiliar environment?

Hypothesis
If rats are exposed to seven days worth of ethanol, then they will show a greater
difference in the behaviors indicative of anxiety between the first and second open field
than the rats that did not receive ethanol.

Methods
Subjects:
20 female long-evans hooded rats, 3 months old at the start of the study. They were kept
on a reverse light schedule and tested during their dark cycle. Food and water were
available at all times. Subjects were randomly assigned to ethanol or control groups by
weight, 12 animals were assigned to the ethanol group, the remaining 8 were controls.
Study design:
All animals were tested in an initial open field prior to ethanol exposure. During open
field testing, rats were placed in the center of a 3 x 3 foot enclosure with 18 inch walls
for 5 minute periods. Above the field, all motion was recorded with a camera and data
was imported to the Anymaze computer program. Ethanol group rats were then given
access to 10% ethanol gel shots with 10% sucrose polymers for seven days and allowed
to self-dose. Control rats were similarly given access to gel that did not contain ethanol.
A second open field was then conducted on the final day of ethanol dosing.

Conclusions

Variable 2: Number of
Line Crossings
Result: p= .27

Variable 1: Time spent in

Center of Open Field
Result: p= .341
There is not enough
evidence to suggest a
difference in time spent in
the center of the open field
between the control
(m=6.00) and ethanol
(m=3.78) groups, t(18)=
-0.978, p>.05

We hypothesized that there would be a greater difference between tests in our three
variables for anxious behaviors for the ethanol group than the control group.
Our results fail to support our hypothesis for time spent in the center of the open field.
We concluded that the difference in the was greater in the control group than the ethanol
group.
Our results fail to support our hypothesis for number of line crossings. We concluded
that the difference was greater in the control group than the ethanol group.
Our results support our hypothesis but are not significant for the time spent grooming.
There was a greater difference in the ethanol group than the control group between the
two tests but the evidence was not significant enough to conclude an effect.

Discussion
We based our hypothesis on the assumption that ethanol would have created a greater
state of anxiety in the rats, producing a larger change in behaviors between the two
tests. However our results did not show a significant variability between the control and
ethanol groups. This drastically differs from the previous studies that we formed our
hypothesis off of.

Change in Line Crossings Between Tests

Future Studies:

There is not enough

evidence to suggest a
difference in number of line
crossings between the
control (m=20.5) and
ethanol (m=15.0) groups,
t(18)= -1.138, p>.05

Control
Linear (Control)
Ethanol
Linear (Ethanol)

Manipulation 1: Remove water sources for the last two days of the gel dosing from both
ethanol and control groups forcing them to consume more of their respective types of
gel. This will theoretically increase the amount of gel consumed before the test which
will increase ethanol intake to create further change in groups.
Manipulation 2: Change the open field test to an elevated plus maze test. A more
complex setting will theoretically help set apart the anxious behaviors between test one
and test two showing how ethanol consumption can affect these behaviors.

For our third test we calculated a chi-square goodness of fit test (alpha level of .05)
between the control and ethanol subject groups to compare the relative frequencies of
the time they spent grooming during the tests.
Change in Time Spent Grooming Between Tests
0-1 sec
1+ sec

Ethanol Rats

42% (5)

58% (7)

Control Rats

50% (4)

50% (4)

Variable 3: Time Spent

Grooming
Result: p= .256373
There is not enough
evidence to suggest that
there are differences in the
proportions of ethanol and
control rats that groomed
for more or less than 1
second, x2 (1, N=20) =
1.2882, p > .05

Sources
1. Colombo, G., Agabio, R., Lobina, C., Reali, R., Vacca, G., & Gessa, G. L. (1998).
Stimulation of locomotor activity by voluntarily consumed ethanol in Sardinian
alcohol-preferring rats. European journal of pharmacology, 357(2), 109-113.
2. Lee, E. H., Tsai, M. J., & Chai, C. Y. (1986). Stress selectively influences center
region activity of mice in an open field. Physiology & behavior, 37(4), 659-662.
3. OCallaghan, M., Horowitz, G. P., & Isaacson, R. L. (1982). An investigation of the
involvement of histaminergic systems in novelty-induced grooming in the mouse.
Behavioral & Neural Biology, 35(4), 368-374. doi: 10.1016/S0163-1047(82)910068.

For Further Information Please Contact:

mmm219@uw.edu
lnb95@uw.edu
cayleigh@uw.edu