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Examining the Inhibitory Effects of LiCl on the Exploratory Behavior and Innate Fear in SpragueDawley Rats in an Open Field

Test
Aisha Gillan
University of Texas at Dallas
ABSTRACT
The purpose of this experiment was to determine if LiCl had any inhibitory effects on the innate
fear activity or exploratory behavior of Sprague-Dawley rats. The rats were injected with either a
1% LiCl solution at 85mgs/kg or a saline solution of equivalent volume and placed in a 4x4 foot
open field test one hour post-injection. Their behavior was observed for 10 minutes and the
amount of line crossings within the apparatus, the number of times they entered the center of
the apparatus and the amount of time they spent in the center was recorded. It was found that
the rats injected with LiCl exhibited less exploratory behaviors than those injected with saline.
The results for the number of time the rats entered the center of the apparatus and the amount
of time they spend there were found to be insignificant. This gave adequate indication that LiCl
does have a detrimental effect on the exploratory behavior of rats.
INTRODUCTION
Previous experiments have shown LiCl to reduce the amount of horizontal and vertical activity in
rats, which can be attributed to lessened exploratory behavior. (Tenk et al 2006) Dosedependent decreases also occurred in locomotor activity following the administration of LiCl. It
has been proven that Lithium Chloride suppresses locomotor activity and ambulation in rats
when they are placed in an open field environment. (Tenk et al 2005) In relation to that, another
study found Lithium Chloride to diminish the curiosity of isolated mice. (Weischer 1979) More

behavioral studies have associated the decrease of ambulatory activity with LiCl administration
with alterations in the rat’s behavior. (Johnson 1976)
In a clinical context, LiCl is used to treat the manic aspect of bipolar disorder. It does this by
decreasing risk-taking behavior. (VanEnkhuizen et al 2015) In this sense, it is thought to alter
behavioral tendencies. This behavioral change is what will be measured in the experiment.
The purpose of this study is to examine the effects of 1% LiCl on the exploratory behavior and
innate fear of Sprague-Dawley rats. One subset of rats (n=8) will be injected with 1% LiCl, while
the other will be injected with saline. One hour post-injection, each rat will be placed in a 4x4
foot empty open box and observed for 10 minutes. Several measures will be taken in order to
quantify their behavioral changes. Their total amount of line crossings, the amount of times they
venture into the center of the open box and the amount of time they spend there will all be
recorded. This study has the potential to show the effects of LiCl on the behavior and riskaverse tendencies of the rats. It is expected that the Lithium Chloride-injected rats will have less
total line crossings, less center entries and less time spent in the center than the control group,
indicating a diminishing effect of LiCl on exploratory behavior and an amplifying effect on their
innate fear.
METHODS
The subjects were 16 male Sprague-Dawley rats weighing approximately 300g. The rats were
housed in cages in a lighted environment prior to injection and were exposed to no loud stimuli.
The rats were administered a 1% LiCl solution at 85mgs/kg or an equivalent volume of saline
intraperitoneally one hour prior to performing the test. This amounted to 2.55mls of a 1%
solution for a rat that weighed 300g. All rats were exposed to the exact same conditions with the
exception of the solution that was injected.

The apparatus was a 4x4 foot open wooden box placed on tile in a dimly lit room. Observations
were carried out by three human researchers who were blind to which rat had been
administered which solution. One researcher measured how many times the rat crossed a floor
tile line, another measured how much total time the rat spent in the center of the apparatus, and
the last measured how many times it ventured into the center of the apparatus.
Each rat was placed in the apparatus for 10 minutes. The apparatus was cleaned with Ethyl
Alcohol after every trial to remove any cues. Behavioral data that was obtained was entered into
and analyzed by Excel. A two-sample T-test was performed assuming equal variances and Pvalues were calculated. Significance was designated as p<0.05.
RESULTS
The effects of LiCl on exploratory behavior in these rats is shown in figure 3, detailed by the
variable of total line crossings. Rats that received the Lithium Chloride had less total line
crossings than the rats that received saline. (91.125 ± 1368.982 vs. 42 ± 807.4286, p = 0.004, t
Stat = 2.9, df = 14, ;Fig. 3). However, the findings do not show a significantly decreased number
of times the experimental rats entered the center of the apparatus compared to the control rats.
(1.75 ± 3.643 vs. 1.375 ± 4.268, p = 0.35, t Stat = 0.377, df = 4 ;Fig. 2). Diminished innate fear
was also measured by the total amount of time the rats spent in the center of the apparatus, but
these results were also found to be insignificant. (6.73 ± 91.355 vs. 3.83 ± 28.663, p = 0.233, t
Stat = 0.7497, df = 14, ;Fig. 1).

8
7

seconds

6
5
4
3
2
1
0
Saline

LiCl

seconds

Figure 1: Mean amount of time (seconds) spent in the center of the apparatus by the LiClinjected rats and the control subject rats.

2
1.8
1.6
1.4
1.2
1
0.8
0.6
0.4
0.2
0
Saline

LiCl

Figure 2: Mean number of times the LiCl-injected rats entered the center of the apparatus
versus the control subject rats.

seconds

100
90
80
70
60
50
40
30
20
10
0
Saline

LiCl

Figure 3: Mean number of total line crossings by the Li-Cl injected rats and the control
subject rats.

DISCUSSION
The results indicate that LiCl does have a measurable effect on the exploratory behaviors of
Sprague-Dawley rats, as seen in the significant decrease of line crossings by LiCl-injected rats.
This present study agrees with similar studies such as Tenk et al 2005 that indicate similar
findings. Unconditioned rats injected with the 1% LiCl solution crossed fewer lines within the
apparatus than the ones injected with saline, suggesting that LiCl has a marked effect on
exploratory behavior. However, the results for the amount of times the rats entered the center of
the apparatus and the amount of time they stayed there were found to be insignificant, showing
no apparent difference in innate fear between LiCl-injected rats and saline-injected rats. These
results contradict the findings in similar experiments, such as in Weischer 1979 and Johnson
1976. The insignificant findings in this study could be attributed to many factors, the first being
that the rats had not been properly habituated prior to experimentation. Their lack of familiarity
with being handled could have caused prior anxiety, leading both subsets of rats to have similar
results in the amount of times they entered the center of the apparatus and the total amount of
time they spent there. This error could also be attributed to the fact that the human researchers

were standing over the apparatus in order to record measures. This could also have caused
prior anxiety in the rats. The findings related to the number of line crossings support the original
hypothesis that LiCl acts to alter behavior in rats. However, in order to make future results more
accurate, rats could spend more time being habituated and measures could be recorded via a
video camera in order to reduce the rodent’s anxiety. In the future, additional experiments could
be performed comparing the behavioral effects of different dosages of LiCl on rats. This would
show the extent to which LiCl has an effect on the rats, and if there is an upper limit to the
response the drug exerts.

REFERENCES
Johnson, F.N. Lithium effects upon components of activity in rats. Specialia (1976): 212-14.
National Center for Biotechnology Information. U.S. National Library of Medicine.
Tenk, Christine M., Martin Kavaliers, and Klaus-Peter Ossenkopp. Dose Response Effects of
Lithium Chloride on Conditioned Place Aversions and Locomotor Activity in Rats.
European Journal of Pharmacology 515.1-3 (2005): 117-27. ScienceDirect.
Tenk, Christine M., Martin Kavaliers, and Klaus-Peter Ossenkopp. The Effects of Acute
Corticosterone on Lithium Chloride-induced Conditioned Place Aversion and
Locomotor Activity in Rats. Life Sciences 79.11 (2006): 1069-080. ScienceDirect.
VanEnkhuizen, Jordy, Morgane Milienne-Petiot, Mark A. Geyer, and Jared W. Young. Modeling
Bipolar Disorder in Mice by Increasing Acetylcholine or Dopamine: Chronic Lithium
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Weischer, Marie-Luis. Influence of Lithium and Rubidium on Exploratory Behaviour and
Locomotor Activity in Isolated Male Mice. Psychopharmacology 61 (1979): 263-66.
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