A PSYCHO-PHARMACOLOGICAL ANALYSIS

OF BEHAVIOUR IN RATS

B.D. GUPTA, P.C. DANDIYA and M.L. GUPTA

Department of Physiology &
Pharmacology, S.M.S. Medical College, Jaipur, India

Received for publication December 1, 1970

The Open Field Test situation is reported to produce autonomic nervous system re
activity determined emotional freezing and defecation (1, 2), cortical excitation determined
rearing activity (3), and corpus striatum activity determined stereotyped ambulation (4,
5). Thus autonomic depressant drugs reduce emotional freezing (6), cortical excitant
drugs augment rearing (7, 8), and hallucinatory drugs facilitate stereotyped ambulation

(4).<BR>
It has been reported (3) that selective stimulation of stereotyped ambulation with
suppression of interrupting responses of rearing or preening is a typical action of increas
ing doses of LSD or iproniazid. This typical drug effect can be differentiated from that
of amphetamine which in increasing doses facilitates rearing and simultaneously blocks

preening as well as stereotyped ambulation; a third type of drug effect being that of imi
pramine which in increasing doses inhibits preening but failing to modify ambulation or
rearing in rats. In the present study, attempts have been made to extend this approach
to other stimulant drugs, i.e. methyl phenidate, pentylene tetrazol, mescaline, caffeine and
pargyline for arriving at some definite conclusions regarding the multidimensional Open
Field performance and the pharmacological manipulation there of.

MATERIALS AND METHODS

Subjects: The subjects of the present investigation were albino rats of 100 to 450g
body weight. The experimental design was composed of 30 groups (6 treatments 5 doses
of each treatment); each group was composed of five male rats. Thus, in all 150 animals
were subjected to the experiment.
Apparatus: The apparatus for the Open Field Test was similar to that used by
Dandiya et al. (4). Briefly, it consisted of an open arena of 323/4inches diameter enclosed
by a wall. The floor was marked out in three concentric circles divided into segments by
lines radiating from the floor centre. The markings provided 25 floor units of approxi
mately equal size. In the Open Field Test, two types of stimuli were presented to the sub
ject, white noise of 78 dB and bright light of 165 footcandles. Hand-operated counters
were used to record the frequency of ambulation, rearing, and preening responses. The
ambulation response was defined as the "walking around" score derived from the number
of radial segments of the arena which were crossed by the subject during the test. Rear
ing response was defined by the number of times the subject stood on its hind limbs.

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Preening score was determined by the number of times the animal scratched its face with
its fore-arms. Defecation score was the number of faecal boluses deposited by the sub
ject on the floor during the test.
Procedure: Double distilled water was used as carrier for all the drugs administered.
According to the body weight of the animal, the required volume of the drug solution or
the distilled water (placebo) was injected intraperitoneally. The concentration of the
drug solutions was such that an animal weighing 250 g may not get more than I ml of the
solution. The doses and the absorption period (latency) of the drugs are listed in Table
1. There were uniform differences between the logarithms of the doses. As soon as
the latency period of the drug elapsed, the treated subiect was carried to the apparatus.
Each subject was tested only for two minutes and scores on ambulation, rearing, preen
ing and defecation were recorded.
TABLE
1. The drugs, dose and the latency period. Each drug group was
composedof 25 rats divided into 5 subgroups. Each subgroup was
given a differentdose (I to V) of the drug intraperitoneallybefore
subjectingthemto the Open Field Test. Figures under columns I
to V are of doses in mg/kg. Latency is the time in hours allowed
betweendrug administrationand subjectingto the Open Field Test.
Placebo was double distilledwater.

RESULTS

A simple analysis of variance was performed on each of the four measures separately.

The analysis of ambulation score showed that the "treatment" effect (F-ratio=21.98:
P<0.01) and its interaction with the "dose" effect (F-ratio=7.52; P<0.01) were produc
ing significant group differences. A breakdown of the "treatment effect" by student's
t-test against the residual variance (Table 2) showed that, except pargyline, each drug
facilitated ambulation as compared with the placebo control; caffeine producing a higher

TABLE 2. The I-test comparisons of the treatment group means on ambulation

scores. N 25.

* Significant at the level of 1% .

 TABLE 3. The t-test comparisons of the treatment group means on rearing scores.
N - 25.

* Significant at the level of 1% .

** Significant at the level of 5%.

TABLE 4. The t-test comparisons of the treatment group means on preening scores.
N-= 25.

* Significant at the level of 1% .

** Significant at the level of 5% .

effect than that of pentylenetetrazol, and methylphenidate or mescaline producing a moder

ate effect on ambulation responses.
The "treatment" effect (F-ratio=58.94; P<0.01) and its interaction with "dose"
effect (F-ratio-5.58; P`0.01) were significant on rearing scores as well. Table 3 shows
the t-test comparisons of the treatment groups with each other. Except pargyline or
mescaline, each of the drugs produced a higher rearing score than that of the placebo
control. The highest frequency of rearing was a function of the methylphenidate group;
the next higher score was that of the caffeine group. Pentylenetetrazol facilitated rearing
moderately.
The significant difference between the groups on the preening responses was the func
tion of the "treatment" effect (F-ratio=22.24; P<0.01) and its interaction with the "dose"

TABLE5. The t-test comparisons of the treatment group means on defecation scores.
N= 25.

* Significant at the level of 1% .

** Significant at the level of 5%.
effect (F-ratio=4.84; P<0.01); Treatment group differences, as measured by t-test and
outlined in Table 4, showed that each of the drugs individually reduced preening perform
ance when compared with the placebo. The highest inhibitory effect was the function
of methylphenidate or pargyline whereas the lowest level of this effect was produced by

pentylenetetrazol; mescaline or caffeine producing a moderate effect of inhibition.

Even on the defecation scores, the "treatment" effect (F-ratio =2.73 ; P<0.01) and
its interaction with "dose" effect (F-ratio=2.54; P<0.01) were found significant in pro
ducing group differences. The treatment group differences, as measured by t-test and
outlined in Table 5, showed that it was only the pentylenetetrazol group which could pro
duce a higher rate of emotional defecation as compared with that of the placebo group.
The "treatment x dose interaction" on all the four parameters of ambulation, rear
ing, preening, and defecation is outlined in the figures.

FIG. 1. Mean ambulation (locomotion) scores FIG. 2. Mean rearing (standing up on hind
of rats subjected to Open Field Test after limbs) scores of rats subjected to Open
administration of five increasing doses (in Field Test after administration of five
logarithms) of methylphenidate (MEP), increasing doses (in logarithms) of mehyl
pargyline (PARG), pentylenetetrazol phenidate (MEP), pargyline (PARG),
(PEN), mescaline (MES) and caffeine pentylenetetrazol (PEN), mescaline(MES)
(CAF). and caffeine (CAF).

DISCUSSION

Taken together, the results of the present experiment have demonstrated that Open

Field behaviour of rats can be modified by the drugs employed in the study. However,

defecation responses is affected only by pentylenetetrazol. Emotional defecation of rats

FIG. 3. Mean preening (scratching of face
with fore-arms) scores of rats subjected
to Open Field Test after administration
of five increasing doses (in logarithms)
of methylphenidate (MEP), pargyline
(PARG), pentylenetetrazol (PEN), mes
caline (MES) and caffeine (CAF).
FIG. 4. Mean defecation (faecal boluses passed
out) scores of rats subjected to Open
Field Test after administration of five
increasing doses (in logarithms) of me
thylphenidate (MEP), pargyline (PARG),
pentylenetetrazol (PEN), mescaline (MES)
and caffeine (CAF).

in the Open Field Test may be a behavioural response of the sympathetic nervous system
over-reactivity (1) and pipradrol (9) or amphetamine (3) fail to augment this response.
Thus, it may be argued that pentylenetetrazol facilitates the defecation responses possibly
due to its overall excitatory effect on central nervous system including those nervous loci
of posterior hypothalamus which control the reactivity of sympathetic nervous system

(10).
Increasing doses of mescaline produce a linear facilitation of ambulation in a fixed

pattern in the outer-most circle of the Open Field arena with a simultaneous inhibition
of interrupting responses of rearing and preening-a behavioural function similar to that
of LSD (4) or of iproniazid (3) in producing this type of stereotyped ambulation (7, 5).
Thus, mescaline produces stereotyped ambulation in contrast to caffeine, methylphenidate
or pentylenetetrazol which, like amphetamine (3), reduce this behavioural pattern; in
creasing doses of these drugs, however, showing a non-linear facilitation of exploratory
ambulation in the Open Field Test. It is interesting to note that a typical pattern of fa
cilitation of rearing with a simultaneous inhibition of preening responses is a function of
these drugs in the following order: methylphenidate, caffeine, and pentylenetetrazol. On
the other hand, pargyline, which fails to affect rearing or ambulation, inhibits preening
behaviour to a larger extent. Thus, as far as the inhibition of preening responses is con
cerned, both central stimulant and anti-depressant drugs have this property; however,
differing from each other in their action on rearing performance. It has been reported
that rearing and preening responses may be behavioural correlates of cortical excitation
and inhibition respectively (3, 7). It may be argued that the central stimulant drugs may
reduce preening by increasing cortical excitation and thereby shifting the "excitation-in
hibition" balance in favour of "excitation" for increasing rearing at the cost of preening.
The anti-depressant drug (pargyline), on the other hand, inhibits preening by directly re
ducing cortical "inhibition"; failing to affect cortical excitation determined rearing.

SUMMARY

In order to find a suitable explanation for the differential behavioural effects of the

pharmacologically similar drugs, several components of the Open Field performance of

rats have been examined under the influence of varying doses of CNS-acting drugs. The
results have shown that facilitation of ambulation with a simultaneous inhibition of the
interrupting responses of rearing and preening is a typical function of increasing doses of
mescaline. On the other hand, facilitation of ambulation and defecation with a simultane
ous blocking of preening is a function of pentylenetetrazol. However, methylphenidate
or caffeine augment ambulation and rearing at the cost of preening in contrast to pargyline
which inhibits preening without affecting the other responses. It is suggested that the
Open Field performance of rats can be used for differentiating cortical stimulant drugs
from the anti-depressant drugs.

Acknowledgement: Authors are thankful to the Indian Council of Medical Research for
providing financial assistance for this study.

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