Behavioural Brain Research 436 (2023) 114085

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Behavioural Brain Research

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Research report

Influence of excessive sucrose consumption on exploratory behaviour in

rats – Possible implications for the brain reward system
Klaudia Modlinska *, Anna Chrzanowska, Katarzyna Goncikowska, Wojciech Pisula
Institute of Psychology, Polish Academy of Sciences, Warsaw, Poland

A R T I C L E I N F O A B S T R A C T

Keywords: Due to the low cost of production and the strong evolutionary preference for sweet taste in humans, sugar is
Exploration added to many food products. This leads to often involuntary overconsumption of high amounts of sugar. Yet,
Reaction to novelty growing evidence indicates that high-sugar diets impact brain function and impair cognitive ability. It may be
Reward brain system
due to physiological changes in specific regions of the brain or/and maladaptive changes in dopamine signalling
Sucrose addiction
similar to those observed in the etiology of addiction. In our study, rats from the experimental group were kept
Hippocampus
Learning on a feeding protocol involving intermittent access to sucrose solution for eight weeks. Then, the animals un­
derwent a spontaneous exploration test in an experimental arena divided into three zones where stationary and
movable objects were placed. Studying the rats’ exploratory behaviour allowed us to assess the impact of the
sucrose diet on a broad spectrum of behaviours related to the general functioning of the organism in its envi­
ronment. Analyses showed differences in reaction to novelty between different diet groups which had been
placed in different experimental setups. Rats from the sugar-fed group responded to change with more pro­
nounced exploratory behaviours directed at the source of the novel stimuli and the surrounding environment.
These results may indicate a lower reward value of novelty resulting from diminished responsiveness of the
reward system in the sugar-diet group. We have not found evidence for memory and/or learning impairments in
rats on the sugar-rich diet.

1. Introduction dopamine signalling similar to those observed in the etiology of addic­

Added sugars, mainly sucrose and high-fructose corn syrup, have
become major components of the human diet today. Sugar consumption
is increasing worldwide (e.g. [1]). This creates a major challenge for
social policy and healthcare systems. The obvious effects of sugar
overconsumption, such as diabetes or obesity, are widely recognised,
while much less attention is paid to cognitive and emotional changes
resulting from a diet based on excessive sugar intake.
Due to the low cost of production and the strong evolutionary pref­
erence for sweet taste in humans, sugar is added to many food products
(including sauces, dairy products, sausage, etc.). This leads to often
involuntary overconsumption of sugar. Yet, growing evidence indicates
that high-sugar diets (HSDs) profoundly impact brain function and
impair cognitive processes even in the absence of extreme weight gain or
excessive energy intake [2,3]. The most important fact is that highly
palatable foods (e.g. products containing sucrose) are likely to increase
dopamine release in the brain reward centres. Over time, regular con­
sumption of this type of food may lead to maladaptive changes in
tion (e.g. [4]). Furthermore, evidence suggests that under certain cir­
cumstances, it can lead to sugar dependency (e.g. [4,5]).
Data from research on rats provide strong evidence that sucrose
consumption can lead to deficits in spatial cognition and reward-
oriented behaviour (for a review, see [6]). These deficiencies were
also observed under conditions of high pre- and postnatal sugar con­
sumption [7,8]. Only a few days of a hypercaloric diet is sufficient to
trigger spatial-specific deficits in rats [2,9]. On the other hand, some
studies suggest that diet-induced spatial memory deficits are not present
when discrete spatial cues are placed in the arena [9]. Such results
indicate that hypercaloric diets may impact different aspects of learning
and memory and that diet-induced behavioural deficits may become
more apparent when task difficulty and cognitive demands are
increased. Another study showed that diet-induced obesity resulting
from excess sucrose intake, but not fat intake, impairs spatial learning
and memory in young animals [10]. Therefore, it is hypothesised that
the effect of sucrose on the brain is not directly correlated with excessive
caloric intake in general.

* Corresponding author.
E-mail address: kmodlinska@wp.pl (K. Modlinska).

https://doi.org/10.1016/j.bbr.2022.114085
Received 2 March 2022; Received in revised form 1 August 2022; Accepted 28 August 2022
Available online 31 August 2022
0166-4328/© 2022 The Author(s). Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
K. Modlinska et al.
The cognitive changes observed in subjects on high-sucrose diets
may be triggered by effects on specific areas of the brain. For example,
the hippocampus, which is involved in spatial, contextual, and episodic
memory formation, is particularly vulnerable to neuropathological
changes following excessive sucrose consumption (e.g., [6,11–16]). In
animal studies, markers of neuroinflammation, such as microglia acti­
vation measured by IBA-1immunoreactivity and increased hippocampal
levels of pro-inflammatory cytokines such as interleukin(IL)-1β, IL6 and
tumour necrosis factor-α (TNF-α), were observed in rats and mice
exposed to HSDs [2,13,14]. Furthermore, high-caloric diets have been
shown to reduce brain-derived neurotrophic factor (BDNF) mRNA
expression in the hippocampus [7,17]. These studies suggest that the
mechanisms through which HSDs impair hippocampal neuroplasticity
may include increases in pro-inflammatory cytokines, as well as de­
creases in neurotrophic factor expression, which may ultimately
contribute to diet-induced cognitive deficits [18]. In rats, HSDs also led
to a decrease in the level of markers of hippocampal neurogenesis in the
dental gyrus [14] and neuroproliferation markers doublecortin and
proliferating cell nuclear antigen (PCNA) immunoreactivity [19].
Another area of research on the influence of excessive sugar con­
sumption on behaviour concentrates on addiction-like changes in the
brain (mainly in the dopaminergic system - for a review, see [4]; also [5,
20,21]). Dependency based on activation of physio-behavioural systems
controlling such areas as metabolism, foraging, and eating to maintain
energy homeostasis is sometimes called “natural addiction” in opposi­
tion to “drug addiction”, which activates specific system related to their
pharmacology [21]. This analogy is based on the propensity of sugar to
evoke bingeing, withdrawal symptoms, and abstinence-induced moti­
vation to gain access to sucrose (e.g., [4]). This motivation occurring
during the deprivation period is manifested in rats, among other things,
by more frequent lever pressing for sugar [22], increased alcohol con­
sumption in rats with sugar-bingeing [23], and stronger response to
sugar-associated cues [24]. In addition, the psychostimulant properties
of sugar are evidenced by the observed cross-sensitization with
amphetamine [21]. From a behavioural perspective, substance depen­
dence may be detected by measuring symptoms of addiction, such as
tolerance (a gradual decrease in responsiveness to the substance
resulting in an increased demand for that substance to produce the same
effect - [25]) or craving (increased efforts to obtain the substance - [4]).
It is well known that repeated activation of the reward circuit by
abusing a specific substance may lead to neuroplastic changes (i.a.
reduction in the density of dopamine receptors). It should be noted that
the dopamine system is highly engaged in memory and learning (e.g.
[26,27]). It also plays a crucial role in initiating motor actions, as
observed in patients with Parkinson’s Disease, and depletion in dopa­
mine in the striatum is associated with disorganization of behaviour [20,
28]. What is more, there is evidence that dopaminergic mechanisms are
critical to the process of cognitive shifting and executive control [27].
Thus, chronic consumption of this type of food may lead to maladaptive
changes in dopamine signalling similar to those observed in the etiology
of addiction [4]. This may be mainly due to a reduction in striatal D2/3
receptor density repeatedly observed in diverse populations of drug
users [29,30], which has also been found in obese subjects [31].
Studies conducted to date indicating adverse changes in cognitive
functioning have focused on cognitive tasks measuring a narrow and
out-of-context range of psychological properties. These included: the
Water Morris Maze (spatial memory and learning), object recognition
test (memory and learning), progressive ratio task (reward processing) -
e.g., [11,31–34]; but: [15,32]. Although these studies show a deterio­
ration in selected cognitive skills in many cases, they cannot show how
these changes translate into the functioning of an individual in a broader
sense. In our project, we intend to identify changes in the overall
behavioural characteristics by using a free exploration protocol [35].
Exploration can be defined as behaviour directed toward acquiring in­
formation about the environment. Among other things, it allows in­
dividuals to prepare for the future (e.g., escape from predators, harsh
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Behavioural Brain Research 436 (2023) 114085
weather conditions, or ensuring food security). It involves a broad range
of behavioural phenomena such as reaction to novelty, risk assessment,
arousal, locomotor activity, habituation, memory, etc. Moreover, a
novel stimulus (even when it lacks rewarding or biologically beneficial
attributes) acts as a positive reinforcer, highlighting its powerful moti­
vational properties (e.g., [36,37]). The organism’s inherent interest in
novelty (the main element of exploratory behaviour) is considered to be
an evolutionary prerequisite for complex learning, and it guides or­
ganisms toward acquiring adaptive behavioural repertoires [38]. On the
other hand, high scores on the novelty-seeking scale are considered risk
factors for several neuropsychiatric disorders, such as addiction and
bipolar disorder [39]. The free exploration test we proposed [35] could
allow us to observe how organisms cope with changes in the environ­
ment in a comprehensive manner, ensuring that the animals’ stress level
remains low and taking into account behavioural dynamics over time.
Unlike most behavioural tests, which measure a single behavioural
parameter, the free exploration test is based on the assumption that
rodents demonstrate their full behavioural spectrum only in a rich
testing environment that enables observation of more complex behav­
iours. From the viewpoint of the theory of levels of integration [40], the
study of exploratory behaviour makes it possible to register changes in
behaviour occurring at higher levels of integration, without being
limited to measuring behaviours or components of behaviour at a lower
level of integration/organization. The general hypothesis underlying
this approach is as follows:
Changes that occur due to sucrose overconsumption may not be fully
manifested in elementary dimensions of behaviour and highly struc­
tured cognitive tasks; they may not exceed the registration threshold.
However, the presence of these changes in various areas of the organ­
ism’s functioning may, as a result of their cumulative effects, lead to
changes at higher levels of integration specific for higher forms of
functioning, e.g., behavioural styles, behavioural syndrome, etc.
Therefore, our research will focus on changes in the overall strategy of
coping with challenges posed by the environment and not on selected
and highly specific aspects of cognitive and emotional functioning (they
will, however, also be controlled throughout the experiment). What is
more, this paradigm seems particularly useful for assessing the influence
of sucrose overconsumption on reward processing (incentive value of
novelty), as well as recognition and memory processing (changes in
experimental settings) associated with the brain reward system and the
hippocampus, respectively.
To increase the accuracy and reliability of the measurement we
decided to provide the test subjects with two levels of novelty as regards
its complexity. It is well known that animals prefer more complex en­
vironments to simplified or impoverished ones [41–44]. This preference
for complexity is most clearly demonstrated when the environmental
change takes the form of an increase in complexity [43]. In the experi­
mental setting, complexity may be introduced in numerous forms. In
addition to placing novel objects in the experimental arena, we can also
provide animals with movable objects. Movability adds a new dimen­
sion to the new object and enables animals to control the novel envi­
ronment to a certain degree. In addition, this specific type of novelty
generates sensory stimuli by virtue of its movability, but also as a result
of the organism’s own activity. Reaction to this kind of stimuli may
differ among individuals with various levels of different psychological
properties (e.g. [45]). Therefore, it is possible to hypothesise that the
expected neuronal and behavioural changes triggered by excessive sugar
consumption may lead to specific reactions to varying levels of
complexity of the novel stimuli.
Considering the above, the general hypothesis underlying our study
is the expectation that high sugar consumption will affect the func­
tioning of the reward system, leading to changes in reaction to novelty
and general exploratory behaviour. Excessive sugar consumption will
induce a tonic, inter-situational increased and persistent desire for
events and stimuli that activate the reward system. The indicators of
these changes will be behaviours focused on novelty seeking, lack of
K. Modlinska et al.
resistance to postponed reinforcements, or high variability in the forms
of activity, combined with an increase in the amount of time spent on the
exploratory activity.
H1. Sugar consumption will lead to increased exploration after the
introduction of novelty. We hypothesise that increased activity
directed at the sources of sensory stimulation will be a result of
decreased sensitivity to the reward value of novelty due to addiction-
like changes in the dopamine system (cf. [39,46,47]).
H2. Sugar consumption will result in a slower rate of habituation to
novelty due to learning impairment resulting from excessive sugar
consumption having a damaging effect on specific areas of the brain
e.g. the hippocampus (cf. [11,31,32]).
H3. Excessive sugar consumption will have a different impact on
reactions to various levels of complexity of the novel stimuli. Higher
complexity (a movable object) will lead to more intensive explora­
tion by stimulating the reward system to a greater degree than a non-
movable object and/or due to a higher level of the reward value of
stimulation resulting from the animal’s own activity (cf. [48]).
To test these hypotheses, we conducted a two-phase study. First, the
animals from the experimental group were maintained on a feeding
protocol involving intermittent access to sucrose solution for eight
weeks. The control group was kept in standard conditions. Second, all
subjects underwent a spontaneous exploration test in an experimental
arena divided into three zones. The test started with a series of habitu­
ation sessions the purpose of which was to familiarise the animals with
the experimental setting. Subsequently, novel objects were placed in one
of the zones, and behavioural reactions to the changed environment
were measured. What is more, half of the animals encountered a novel
stimulus with increased complexity (i.e. a movable object). We believe
that this procedure allowed us to induce symptoms of sugar dependency,
making it possible to assess a broad range of behavioural changes
resulting from the impact of sucrose on its neural substrates.
2. Methods
2.1. Animals
The sample consisted of 49 male Lister Hooded rats. The rats were
bred and housed in the vivarium of the Institute of Psychology, Polish
Academy of Sciences, Warsaw, Poland. At the beginning of the study, the
rats were approx. 30 days old and weighed approx. 140 g.
The rats were housed in groups of 3–4 in Tecniplast© Eurostandard
Type IV cages (610 mm × 435 mm × 215 mm) with dust-free softwood
granules Tierwohl Super© as bedding. They had ad libitum access to
water and standard laboratory fodder (Labofeed H, WP Morawski,
Kcynia, Poland). The day/night cycle was set at 12/12 h (lights-on at
8.00 a.m.). The temperature was maintained at a constant 21–23ºC, and
humidity at 45–60 %. Prior to the experiment, the cages were cleaned
once a week. However, in order to ensure that the experimental pro­
cedure was not disturbed, the cages were cleaned just before the start of
the behavioural test and again after the test was completed.
All the rats were housed, bred and taken care of in accordance with
the Regulation of the Polish Minister for Agriculture and Rural Devel­
opment of 14 December 2016 on laboratory animal care. The experi­
mental procedures were approved by the First Local Committee for
Ethics in Animal Experimentation in Warsaw, Poland, permit #1268/
2021.
2.2. Feeding protocol
Prior to the experiment, the animals followed two different diet
protocols for a period of 8 weeks. One group was given access to sucrose
solution for 2 h per day at the beginning of the circadian cycle’s dark
(active) phase. During the access to the sucrose solution, no plain water
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Behavioural Brain Research 436 (2023) 114085
was available to the animals. Sucrose is the most commonly consumed
sugar-based sweetener (a disaccharide containing 50 % fructose and 50
% glucose). The animals were given a 10 % sucrose solution, which was
similar to the sugar dose found in commercially available sweet bever­
ages and in diets with hidden sugars [49,50]. The Control group fol­
lowed the standard feeding protocol with ad libitum access to plain
water.
Bottles with sweetened water were weighed before and after placing
them in the rats’ home cages. Analysis of mean values of a weekly su­
crose solution consumption using Anova indicated that the amount of
sugar water drank by rats increased significantly during the 8-week
feeding procedure (F(7,35) = 16.474; p < 0.001) - Fig. 1. There were
no differences in body weight between the groups at the start of the
behavioural test (approx. 430 g for an animal).
2.3. Experimental setup and procedure
The measurement apparatus and methods were similar to those used
in our previous studies [35,43,51–54]. The experimental chamber
(Fig. 2) was a box measuring 800 mm × 600 mm × 800 mm. The
chamber was divided into three zones: A, B, and C by two walls running
perpendicularly to its longer side. The partition walls between the zones
had triangular openings (120 mm × 140 mm) at the bottom, which
enabled free movement between the chamber parts. There was a hole
curved in the back wall of the chamber that served as an entrance for
animals moving from the transporter into the chamber. The front of the
chamber was made of transparent plexiglass and it could be lifted to
obtain full access to the experimental arena. The entire chamber was
covered with a layer of washable varnish. In zones B and C, we placed
tunnels (200 mm × 120 mm × 80 mm) made of hardwood covered
with washable paint. In contrast to the most frequently used
two-dimensional experimental settings, these tunnels provided a com­
plex three-dimensional environment. The central zone (A) was left
empty.
At the start of each trial, a transporter (a small cylindrical cage –
60 mm in diameter with doors 120 mm high and 100 mm wide) with the
tested animal inside was placed by the entrance to zone A (Fig. 2) behind
the wall of the experimental chamber. The entrance door was then lifted,
and it was left open until the end of the trial. The animal was free to stay
in the transporter or leave it to explore the chamber. The first seven
trials were habituation trials during which the apparatus was arranged
in the same way. The introduction of novelty (addition of tunnels) took
place between trials 7 and 8. The three subsequent trials were conducted
with the chamber in this new arrangement (Fig. 3). Each trial was 7 min
long and was conducted for each animal once a day.
Two different types of novelty were introduced in trial 8. In the first
setting, the novelty was introduced by adding new tunnels to the
experimental arena. The second setting had the same configuration of
added tunnels, but the top tunnel was built in a way allowing the rats to
Fig. 1. Sucrose solution consumption (in grams per day) in the sugar-diet group
prior to the behavioural test.
K. Modlinska et al. Behavioural Brain Research 436 (2023) 114085

Fig. 2. Experimental chamber - frontal view through the transparent front wall.

Fig. 3. Arrangement of objects in the experimental chamber in each experimental setting.

move it like a seesaw (Fig. 3).
Setting 1 - Addition of a stationary novel object in the experimental
box (Add group). During the habituation sessions, two tunnels
(200 mm × 120 mm × 80 mm) were present in each of zones B and C.
They were arranged in the same manner (Fig. 2). On the first experi­
mental day (trial 8), two supplementary tunnels were placed in zone C
(Fig. 3). The arrangement of the tunnels in zone B remained unchanged.
A total of 27 (12 rats from the sugar diet and 15 rats from the standard
diet) rats were used for this condition.
Setting 2 - Addition of a movable novel object in the experimental
box (MoveAdd group). During the habituation sessions, two tunnels
(200 mm × 120 mm×80 mm) were present in each of zones B and C.
They were arranged in the same manner (Fig. 2). On the first
experimental day (trial 8), two supplementary tunnels were placed in
zone C (Fig. 3). The tunnel located on top resembled a seesaw – it was
the same structure as in the other configurations, but it was placed on
four small pegs (which served as pivots) in the middle and at the end of
the side walls. The pegs allow for movement only in one direction. This
meant that when one end of the tunnel was touching the ‘roof’ of the
tunnel underneath it, the other end was raised. The arrangement of the
tunnels in zone B remained unchanged. A total of 35 (12 rats from sugar
diet and 23 rats from standard diet) rats were used for this condition.
To avoid the deceptive effect of lateralization or visual-auditory
cues, the novelty was introduced into the right zone (as described
above - zone C) for half of the rats and in the left zone (zone B) for the
remaining half (mirror image of Fig. 3).

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K. Modlinska et al. Behavioural Brain Research 436 (2023) 114085

2.4. Data processing and statistical analyses

We used BORIS software [55] to encode the behaviours on the basis

of the recorded material, which made it possible to define selected be­
haviours and to assess their duration and frequency. We scored the be­
haviours the animals engaged in during the entire experimental trial.
Consequently, we were able to assign specific scores to the time of
separate bouts of behaviours, their frequency, and the total time the
animals spent engaging in a given behaviour. The following variables
were measured: (1) Time spent in the transporter (excluding the latency
to leave the transporter); (2) Time spent in the unchanged zone of the
chamber; (3) Time spent in the changed zone of the chamber; (4) Fre­
quency of moving between the zones (left/right/transporter) of the
chamber; (5) Time spent on contact with the tunnels in the unchanged
zone of the chamber; (6) Frequency of contact with the tunnels in the
unchanged zone of the chamber; (7) Time spent on contact with the
tunnels in the changed zone of the chamber; and (8) Frequency of
contact with the tunnels in the changed zone of the chamber.
To enhance the legibility of the results and tables, the habituation
phase has been indicated as H, while the test trials have been indicated Fig. 4. Z-scores for the time spent in the transporter in groups encountering
as T1, T2, and T3, respectively. Novelty (i.e. addition of tunnels) was different types of novelty.
introduced in the first test trial (T1). Different experimental settings (a
movable or stationary tunnel as novel objects) are indicated in the Re­ p = 0.034; T3: t = 3.833; p = 0.005).
sults as a “move” factor.
The first step of the analysis was to establish a reference value for the
3.2. Time spent in the changed zone of the chamber
test trials. The mean value for the habituation trials H5 to H7 was
calculated, and the created variable was labelled as ‘habituation phase’
Mauchly’s test indicated that the assumption of sphericity had been
(phase H). The next step consisted in calculating the Z-scores for phase H
violated (χ2(5) = 17.884, p = 0.003), so the degrees of freedom were
within the four experimental samples separately. Next, all values from
corrected using Greenhouse-Geisser estimates of sphericity (ε = 0.797).
test trials T1 to T3 were converted to Z-scores on the basis of phase H
The analysis showed an interactive effect of trial and diet: F
statistics (Mean, StdDev). These data were analysed using a General
(2.391,107.604)= 5.010, p = 0.005, Eta2 = 0.100. The following main
Linear Model procedure (GLM), with repeated measurements (H, T1, T2,
effects were also detected: a main effect of diet: F(1,45) = 11.293,
T3) as within-subject factors, as well as diet and setting assignments as
p = 0.002, Eta2 = 0.201, and a main effect of trial: F(2.391,107.604)
between-subject factors. This was followed by PostHoc t-tests with
= 60.220, p < 0.001, Eta2 = 0.572. There was no interaction effect of
Bonferroni correction for multiple comparisons. Differences were
trial, group and diet or an interactive effect of move and diet or an
considered significant for p ≤ 0.05.
interactive effect of trial and move or a main effect of the move.
In addition, partial eta squared (Eta2) obtained in the RM ANOVA
A post hoc analysis showed that rats on different diets demonstrated
were analysed using Kruskal-Wallis ANOVA to compare the effect size of
different patterns of reaction to change in the experimental box – Fig. 5.
different factors under study. Despite some limitations, partial eta
Rats maintained on the standard diet increased the time spent in the
squared (Eta2) is regarded as an effect size measure, which makes it
changed zone in the first test trial compared to the habituation phase
possible to compare the results obtained in different subject groups or
(T1: t = 6.191; p < 0.001). They also spent more time in this zone in the
even studies. Therefore, we decided to use this measure in order to
second (T2: t = 4.315; p < 0.001) and third test trials (T3: t = 6.194;
obtain more detailed information about the effects of experimental
p < 0.001) compared to the habituation phase. There were no differ­
factors on rat behaviour in the experimental box. For non-significant
ences in the time between test trials. However, in rats from the sugar-fed
effects, the Eta2 value was set to “0”.

3. Results

3.1. Time spent in the transporter

The analysis showed an interactive effect of trial and move: F

(3135)= 4.429, p = 0.005, Eta2= 0.090; a main effect of trial: F
(3135)= 14.041, p < 0.001, Eta2= 0.238; and a main effect of move: F
(1,45)= 18.637, p < 0.001, Eta2= 0.293. There was no interaction effect
of trial, move and diet or effect of trial and diet or diet and move. There
was no main effect of diet either.
A post hoc analysis showed that rats from different groups followed
different patterns of reaction to change in the experimental box - Fig. 4.
In the setup with added stationary tunnels, there was a decrease in the
time spent in the transporter in the first test trial (T1: t = 6.151;
p < 0.001). Then the time remained low. In the condition with the added
movable tunnel, there was no change in the time spent in the transporter
(p > 0.05). Differences between the groups were also observed in indi­
vidual trials. Rats from the added stationary tunnel setup spent less time
in the transporter than their counterparts from the added movable Fig. 5. Z-scores for the time spent in the changed zone of the chamber in the
tunnel setup in all test trials (T1: t = 3.935; p = 0.003; T2: t = 3.298; sugar-fed and standard-diet groups.

5
K. Modlinska et al.
group, after an increase of the time spent in the changed zone in the first
test trial (T1: t = 11.382; p < 0.001), there was a decline of the time in
the second test trial (T2: t = 5.414; p < 0.001). Then, the time remained
constant in the third test trial. There were also differences in the time
spent in the changed zone between the diet groups. Rats from the sugar-
fed group spent more time in that zone than rats from the standard-diet
group in the first test trial (T1: t = 4.737; p < 0.001).
3.3. Time spent in the unchanged zone of the chamber
Mauchly’s test indicated that the assumption of sphericity had been
violated (χ2(5) = 21.726, p < 0.001), so the degrees of freedom were
corrected using Greenhouse-Geisser estimates of sphericity (ε = 0.807).
The analysis showed an interactive effect of trial and diet: F
(2.422,108.987) = 3.947, p = 0.016, Eta2 = 0.081, a main effect of diet:
F(1,45) = 24.349, p < 0.001, Eta2 = 0.351, a main effect of move: F
(1,45) = 11.075, p = 0.002, Eta2 = 0.198, and a main effect of trial: F
(2.422,108.987) = 12.155, p < 0.001, Eta2 = 0.213. There was no
interaction effect of trial, group and diet or effect of trial and group.
A post hoc analysis showed that rats on different diets demonstrated
different patterns of reaction to change in the unchanged zone of the
chamber – Fig. 6. Rats maintained on the standard diet spent less time in
the unchanged zone in the first test trial compared to the habituation
phase (T1: t = 4.406; p < 0.001). The time remained low in the subse­
quent test trials. However, in rats from the sugar-fed group, time spent in
the unchanged zone in the first test trial was the same as in the habit­
uation phase (T1: p > 0.05). This time then decreased in the third test
trial compared to the second trial (T3: t = 4.047; p = 0.002). There were
also differences between the diet groups in the time spent in the un­
changed zone. Rats from the sugar-fed group spent more time in the
unchanged zone than rats from the standard-diet group in the first test
trial (T1: t = 3.716; p = 0.008), and in the second test trial (T2:
t = 4.228; p = 0.001).
In addition, there was a difference between the rats encountering
different types of novelty. In general, rats from the added movable
tunnel setup spent more time in the unchanged zone than rats from the
other setup (t = 3.328; p = 0.002).
3.4. Frequency of moving between the chamber zones (left/right/
transporter)
The analysis showed an interactive effect of trial and diet: F(3135)=
8.055, p < 0.001, Eta2 = 0.152; an interactive effect of trial and move: F
(3135) = 11.212, p < 0.001, Eta2= 0.199; and a main effect of trial: F
Fig. 6. Z-scores for time spent in the unchanged zone of the chamber in the
sugar-fed and standard-diet groups.
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Behavioural Brain Research 436 (2023) 114085
(3135) = 16.342, p < 0.001, Eta2 = 0.266, a main effect of diet: F(1,45)
= 5.123, p = 0.028, Eta2 = 0.102, and a main effect of move: F(1,45)=
6.032, p = 0.018, Eta2 = 0.015. There was no interaction effect of trial,
group and diet and no interactive effect of diet and move.
A post hoc analysis also showed that rats on different diets demon­
strated slightly different patterns of behaviour – Fig. 7. Rats maintained
on the standard diet moved between the zones less frequently in the first
test trial (T1: t = 4.351; p < 0.001), and less frequently in the third test
trial (T3: t = 5.240; p < 0.001) compared to the habituation phase. The
frequency increased in the second test trial (T2: t = 3.240; p = 0.042)
compared to the first test trial, and again decreased in the third test trial
(T3: t = 4.130; p = 0.002). In rats from the sugar-fed group, the fre­
quency of moving between the zones decreased in the first test trial (T1:
t = 4.008; p = 0.003). Then, the frequency increased in the second test
trial (T2: t = 5.238; p < 0.001), and then remained the same in the third
trial (p > 0.05). There was a difference in the frequency of moving be­
tween the zones in the third test trial between the diet groups. Rats from
the sugar-fed group move between the zones less frequently than rats
from the standard-diet group (T3: t = 4.735; p < 0.001).
A post hoc analysis showed that rats from different conditions also
demonstrated different patterns of behaviour – Fig. 8. In the setup with
added stationary tunnels, there was a decrease in the frequency of
moving between the zones in the first test trial (T1: t = 5.923;
p < 0.001), then an increase in the second test trial (T2: t = 6.317;
p < 0.001), and again a decrease in the third test trial (T3: t = 5.727;
p < 0.001). In the setting with the added movable tunnel, there was no
change in the test trials compared to the habituation phase (T1:
p > 0.05). However, a change between the first and third test trials was
observed (t = 3.560; p = 0.014), with higher frequency in the third test
trial. A comparison of the frequency levels in the individual trials indi­
cated that rats from the added movable tunnels setting moved between
the zones more frequently than rats from the other setup in the third test
trial (T3: t = 5.038; p < 0.001).
3.5. Time spent on contact with the tunnels in the changed zone of the
chamber
Mauchly’s test indicated that the assumption of sphericity had been
violated (χ2(5) = 19.001, p = 0.002), so the degrees of freedom were
corrected using Greenhouse-Geisser estimates of sphericity (ε = 0.822).
The analysis showed an interactive effect of trial, diet and group: F
(2.467,111.015)= 3.926, p = 0.016, Eta2= 0.080; an interactive effect
Fig. 7. Z-scores for the frequency of moving between the chamber zones in the
sugar-fed and standard-diet groups.
K. Modlinska et al.
Fig. 8. Z-scores for the frequency of moving between the chamber zones in
groups encountering different types of novelty.
of trial and diet: F(2.467,111.015)= 3.894, p = 0.016, Eta2= 0.080; an
interactive effect of trial and move: F(2.467,111.015)= 3.820,
p = 0.018, Eta2= 0.078; an interactive effect of diet and move: F
(1,45)= 16.795, p < 0.001, Eta2= 0.272; a main effect of diet: F(1,45)=
9.428, p = 0.004, Eta2= 0.173, a main effect of move: F(1,45)= 15.804,
p < 0.001, Eta2= 0.260, and a main effect of trial: F(2.467,111.015)=
62.019, p < 0.001, Eta2= 0.580.
Post hoc analyses showed differences in reaction to change between
different diet groups subject to different conditions during the
Fig. 9. Z-scores for the time spent on contact with the tunnels in the changed zone of the chamber in the sugar-fed and standard-diet groups encountering different
types of novelty.
7
Behavioural Brain Research 436 (2023) 114085
experiment – Fig. 9. Rats from the group maintained on the standard diet
spent more time exploring the added stationary tunnels in the first test
trial (T1: t = 4.112; p = 0.008), and that amount of time remained high
in consecutive test trials. A similar pattern was observed in the rats
maintained on the standard diet which encountered the added movable
tunnel (T1: t = 5.550; p < 0.001). There were no differences between
these two groups in the individual trials.
Although in rats maintained on the sugar diet there was also an in­
crease in the time spent on contact with the tunnels in the first test trial
in the setting with stationary tunnels (T1: t = 10.600; p < 0.001), as well
as in the setting with movable tunnels (T1: t = 5.479; p < 0.001), there
were differences in the amount of time between individual trials. Rats
from the group confronted with a stationary tunnel spent more time
exploring the tunnels in the changed zone than rats which encountered a
movable tunnel in the first (T1: t = 4.621; p < 0.001), second (T2:
t = 4.745; p < 0.001), and third test trials (T3: t = 4.743; p < 0.001).
In addition, rats from the sugar-fed group spent more time exploring
the stationary tunnels than rats from the standard-diet group in the first
(T1: t = 5.855; p < 0.001), and in the second test trial (T2: t = 3.759;
p = 0.028). There were no differences between the diet groups in the
see-saw-tunnels setup.
3.6. Time spent on contact with the tunnels in the unchanged zone of the
chamber
The analysis showed only a main effect of trial: F(3135)= 6.713,
p < 0.001, Eta2= 0.130, and a main effect of move: F(1,45)= 14.621,
p < 0.001, Eta2= 0.245.
In general, rats from the added movable tunnel setup spent more
K. Modlinska et al.
time on contact with tunnels in the unchanged zone than rats from the
other setup (t = 3.824; p < 0.001).
3.7. Frequency of contact with the tunnels in the unchanged zone of the
chamber
The analysis showed an interactive effect of trial and diet F(3135)=
3.539, p = 0.016, Eta2= 0.073, a main effect of trial: F(3135)= 4.072,
p = 0.008, Eta2= 0.083, and a main effect of diet: F(1,45)= 16.102,
p < 0.001, Eta2= 0.264, and a main effect of move: F(1,45)= 6.588,
p = 0.014, Eta2= 0.128. There was no interaction effect of trial, move
and diet or effect of trial and move. No interactive effect of move and
diet was found either.
A post hoc analysis showed differences between the rats on different
diets in terms of their reaction to change in the experimental box. Rats
maintained on the standard diet interacted with the tunnels in the un­
changed zone in the third test trial less frequently than in the habitua­
tion phase (T3: t = 3.576; p = 0.014). There were no differences
between the first and second test trials or between the second and third
trials. In rats maintained on the sugar-rich diet, there were no changes in
the frequency of interactions with tunnels in the unchanged zone
(p > 0.05) throughout the experiment. The diet groups differed in terms
of frequency of contact with the tunnels across individual trials. Rats
from the sugar-fed group demonstrated a higher frequency than rats
from the standard-diet group in the second test trial (T2: t = 3.610;
p = 0.011) and the third test trial (T3: t = 3.773; p = 0.006).
Moreover, rats from the group which encountered the movable
tunnels had contact with the tunnels in the unchanged zone more
frequently than the rats from the added stationary tunnel setup
(t = 2.567; p = 0.014).
3.8. Frequency of contact with the tunnels in the changed zone of the
chamber
The analysis showed only a main effect of the trial: F(3135)= 7.126,
p < 0.001, Eta2= 0.137. There was no interaction effect of trial, group
and diet or effect of trial and group or effect of trial and diet. No main
effects of group or diet were found either.
A post hoc analysis showed a significant increase in the frequency of
interaction with the tunnels in the changed zone in the third test trial
(T3: t = 4.608; p < 0.001) compared to the habituation phase. There
were no changes between the first, second and third test trials.
3.9. Effect size analysis
Kruskal-Wallis ANOVA showed (H=22.004, df=6, p < 0.001) that
the effects associated with the experimental factors ranked differently,
with the Trial factor ranking first, followed by Move and Diet factors.
Table 1 shows the median of Eta2 for experimental effects.
3.10. Summary of results
Rats maintained on the sugar diet differed substantially from rats
maintained on the standard diet in terms of their reaction to change
introduced in the experimental chamber. Subjects from the sugar-fed
Table 1
Median of Eta2 for experimental effects.
Group N Median
Trial 8 0.225
Move 8 0.158
Diet 8 0.137
Trial x diet 8 0.08
Diet x move 8 0.0
Trial x move 8 0.0
Trial x move x diet 8 0.0
8
Behavioural Brain Research 436 (2023) 114085
group increased their activity in the zone where the change was intro­
duced in the first test trial, but then the activity decreased and remained
at a low level in the third test trial. On the other hand, subjects from the
standard-diet group increased their activity after the introduction of
novelty, but the activity remained high until the end of the experiment.
All groups of rats increased their contact with the tunnels in the changed
zone after new tunnels had been introduced. However, rats from the
sugar-fed group spent more time exploring the stationary tunnels than
investigating the movable tunnels across all test trials. In addition, the
sugar-fed group spent more time exploring the stationary tunnels
compared to the standard-diet group (in the first and second test trials).
In addition, the increase in the duration of contact with the tunnels after
the introduction of novelty was much higher in the sugar-fed group than
in the standard-diet group (in the stationary tunnels setting).
Differences were also observed in the unchanged zone of the box.
Animals from the standard-diet group decreased the amount of time
spent in this zone after the novelty had been introduced in the other
zone, while their activity remained low in the subsequent trials. Animals
from the sugar-fed group maintained the same level of activity in the
unchanged zone despite the introduction of novelty in the other zone. In
general, rats from the sugar-fed group spent more time in the unchanged
zone than the rats from the standard-diet group (in the second and third
test trials). Moreover, the frequency of contacts with the tunnels in the
unchanged zone was higher in rats from the sugar-fed group (in the
second and third test trials).
Rats from different diet groups differed slightly in terms of the fre­
quency of moving between the chamber zones. Both groups decreased
the rate of moving between the zones in the first test trial and then
increased it in the second trial, but the animals from the control group
again decreased the frequency in the third trial, while the rats from the
sugar-fed group maintained the same frequency as in the second test
trial.
Furthermore, rats from the stationary tunnels setting reduced their
frequency of moving between the zones after the introduction of the
novel tunnels then increased the frequency in the second test trial and
decreased it again in the last test trial. Rats from the movable tunnels
setting did not change their behaviour in this respect after the intro­
duction of novelty, but the frequency in the third trial was higher than in
the first trial.
There was also a difference in the amount of time spent in the
transporter between the experimental setups. Rats from the stationary
tunnels group decreased the amount of time spent in the transporter in
the first trial and generally spent less time in the transporter than the rats
from the other setup.
The effect size analysis has shown that the experimental factors
directly linked to the test arena arrangement (Trial and Move) play a
major part in explaining the variability in the rats’ behaviour. This
seems in line with current knowledge about environmental control over
behaviour. The Diet factor was placed in the third position, with the
median Eta2 = 0.137. The Diet factor, together with the interaction of
Diet and Trial factors, explained more than twenty percent of behaviour
variance (Eta2 = 0.217), which should be considered to be a powerful
effect.
4. Discussion
Studying intrinsically motivated exploratory behaviour makes it
possible to formulate hypotheses about the effects of experimental
manipulation on reward-mediated neural pathways. This approach is
based on the assumption that neophilic preferences, which are the main
feature of exploration, reflect the reward value of novelty [37]. The
mesolimbic dopamine system plays a major role in the processing of the
reward value of novelty, as it does for drugs of abuse and other forms of
reinforcement [56]. Novel stimuli are known to excite dopamine neu­
rons [57] and boost signalling in brain regions receiving dopaminergic
input [58]. Therefore, the influence of factors affecting the reward
K. Modlinska et al.
system may well be detected in the analysis of reactions to novelty (cf.
[59]). As mentioned in the Introduction, excessive sugar consumption
may lead to changes in the dopamine system similar to those observed in
drug addiction (e.g. [4]). We hypothesised that the influence of a
sugar-rich diet on the brain reward system may be assessed by analysing
exploratory behaviour.
The results of our study showed differences in reactions to novelty
between different diet groups in different experimental settings. It is
known that the introduction of novelty into a familiar environment
triggers a range of investigative behaviours directed at the source of the
novel stimuli [37]. In our experiment, the animals maintained on a
sugar-rich diet responded to the change in the experimental box by
spending three times more time exploring the novel stimulus than the
animals from the standard-diet group. The amount of time spent in the
changed zone of the experimental arena was also much higher but
contrary to the standard-diet group, which spent more time in the
changed zone throughout the consecutive sessions, the rats from the
sugar-fed group spent less time in that zone in the next test session and
maintained this lower level in the third test trial. These results may
indicate a lower reward value of novelty in the sugar-fed group, which
needed to spend more time on exploration in order to reach the same
level of reward as the other group. This argument is supported by the
fact that the diminished responsiveness of the reward system was
detected in the pre-test period when the rats kept on the sugar-rich diet
increased their sugar intake in the course of eight weeks. In addiction
studies, this reaction is considered to be a sign of tolerance to substances
consumed in excessive amounts and cravings (e.g. [25], for a review, see
[4]). This strong reaction to novelty may also suggest a
cross-sensitisation effect of sugar, eliciting a stronger response to the
rewarding stimulus (cf. [60,61]). However, sensitisation often involves
increased motor activity (e.g. [4]), which was not observed in our study
as measured by the frequency of moving between the zones of the
experimental box. Both groups demonstrated similar levels as regards
this behaviour until the last test trial when the rats from the
standard-diet group reduced the frequency of moving between the zones
of the experimental arena, but the rats from the sugar-fed group
exhibited a sustained high frequency of this behaviour. This effect,
however, seems to be consistent with the overall higher demand for
stimulation in the sugar-fed group.
Enhanced reactions to the introduction of novelty may also be
explained in terms of heightened impulsivity in the sugar-fed group (cf.
[62]). Impulsive behaviours in chronic drug abusers have been found to
be associated with reduced dopamine D2/3 receptors in the striatum (e.
g. [63]), and a low D2/3 receptor availability was correlated with lower
latency to come into contact with the novel object, which is consistent
with higher novelty-reactivity and potentially higher impulsivity [64].
Again, these insights point to the dopaminergic system being involved in
establishing differences in behaviour between the different diet groups.
Reaction to novelty is not the only element of exploratory behaviour
that can be empirically observed. As proposed by Berlyne in his classical
work [65], exploration involves not only specific exploration directed
towards a specific source of stimulation but also diversive exploration
comprising responses directed at a wide range of stimuli. This type of
behaviour enables an ongoing revision of mental maps and enhanced
certainty as to the information obtained from the environment, which
seems to be a crucial adaptive function [66]. In our study, rats from the
sugar-fed group reacted to the introduction of novelty with a high rate of
object exploration, but at the same time, they did not reduce the time
spent in the unchanged zone of the experimental box and maintained a
relatively high rate of contacts with the unchanged tunnels in the test
trials. This may indicate a higher level of diversive exploration in this
group, suggesting a higher demand for updated information about the
environment. Although the neural substrates of this distinctive type of
exploration are yet to be identified, it may be hypothesised that the
hippocampal regions involved in memory processing are employed.
Based on our findings, it is unclear if this behaviour relates to memory
9
Behavioural Brain Research 436 (2023) 114085
malfunction in sugar-fed rats. It is also likely that this kind of “broad”
exploration constitutes, to some degree, a source of reward to the ani­
mals and that it is influenced by the same changes in the reward system
as the reaction to the object’s novelty. Some support for this point of
view is provided by Ricker and colleagues [67], who claim that a
self-directing choice behaviour can lead to the choice itself becoming a
reinforcing factor.
However, these findings hold true only with regard to the stationary
tunnels’ setting. Novelty may also involve changes in the complexity of
the environment, with more complex objects eliciting a stronger
exploratory reaction [43,54]. The magnitude and scope of this reaction
depend on the degree of change in the environment. It is characterised
by the discrepancy between a single exposure to a stimulus and subse­
quent exposures. Therefore, an increase in complexity should lead to
increased exploration. In our study, the introduction of a movable tunnel
did not elicit an enhanced exploratory response in the sugar-fed group,
and the rats’ activity was similar to the activity of the standard-diet
animals. It is possible that the movable object (placed on top of the
two-level tunnel structure and moving unexpectedly like a seesaw when
a rat interacted with it or climbed on top of it) may have been a source of
stress. Even though the animals could recognise the stimulus as novel,
they may have chosen not to approach or investigate it. Neophobia (fear
of novelty) can suppress neophilia if the degree of novelty is too high or
if some other factors generate high levels of fear, as demonstrated in the
theoretical analysis by R. Hughes [37]. It seems that this fear reaction
was more pronounced in the sugar-diet group, resulting in the reward
value of novelty being reduced. The reasoning about the fear response to
a movable novel object is also supported by the differences in time spent
in the starting box between the groups encountering different types of
novelty. The starting box is a safe, familiar environment, and the time
spent in it may be considered a measure of anxiety (cf. [68]). In our
study, the rats exposed to novelty in the form of a seesaw tunnel spent
more time in the starting box in all test sessions, which may suggest that
this change increased the level of neophobia. However, the fear response
hypothesis does not support the earlier assumption about increased
impulsivity in the sugar-fed group. Stressful situations should lead to
more impulsive behaviours and higher novelty/sensation seeking in this
group (cf. [69]).
Another aspect of neural changes resulting from excessive sugar
consumption, which can be discussed by analysing exploratory behav­
iour, is memory processing. Numerous studies point out to the hippo­
campus as the region significantly affected by sugar-rich diet (e.g., [6,
11–16]. It has been suggested that relationships between hippocampal
memory processes and neotic preferences may specifically involve
novelty detection in which current experiences are compared with
encoded information about past events [70–72]. If such comparisons
result in a discrepancy between the stored data and the current event,
the detection of novelty would be registered and addressed with specific
behaviour [73]. Hippocampus impairment may lead to interference with
information comparison and thus with the ability to detect or respond to
novelty [74,75]. Another sign of properly functioning memory is the
relatively quick pace of habituation to change. Our analyses have not
shown any negative changes in memory processing. Habituation to the
experimental setup was similar in animals from both diet groups, as
measured by the reduction in the time spent investigating the novel
tunnels in the successive test trials. The rapid decrease in time spent in
the changed zone of the experimental arena seems to support this
conclusion. It is not necessarily in conflict with previous findings, as the
memory functions of the hippocampus are mainly associated with its
dorsal region, in contrast to the regulation of fear or anxiety-like
response associated with the ventral region of the hippocampus [75].
It may be speculated that different regions of the hippocampus respond
differently to excessive sugar consumption (cf. [76]). However, not all
studies on the effect of sucrose consumption have detected impairments
in all memory functions [77,78]. What is more, some findings indicate
that sucrose overconsumption may lead to improved learning in the
K. Modlinska et al. Behavioural Brain Research 436 (2023) 114085

operant bar-pressing task [79], which may be associated with a higher
reward value in sugar-fed subjects.
To sum up, the results of our study seem to corroborate previous
findings about the significant influence of intermittent sucrose con­
sumption on the brain reward system. Despite a subtle manipulation of
the feeding protocol, which can be roughly compared to humans eating
a few sweets every day, the observed behavioural changes in the
experimental group seem very significant. As illustrated by effect size
analyses, the diet factor is responsible for over 20 % of the behavioural
variance. All animals were kept in very similar conditions. There were
no differences in the animals’ weight and overall characteristics. It
looks, therefore, that even in the test situation, where the proximate/
situational factors play major roles, subjects following the sugar-rich
diet behaved significantly differently compared to their counterparts
maintained on the standard rat diet. On the other hand, we have not
found any evidence for memory and/or learning impairments in rats on
a sugar-rich diet. It may be suggested that the dosage of sucrose pro­
posed in our feeding scheme was too low or that sucrose consumption
has a limited damaging effect on the hippocampus. Nonetheless,
studying exploratory behaviour allows us to assess the impact of a
sucrose-rich diet on a broad spectrum of behaviours related to the
general functioning of the organism in its environment. Our study has
contributed to the existing body of knowledge about the influence of diet
on cognitive and emotional development.
Yet, the presented above analyses of results have some important
limitations. Our study aimed to pinpoint the relation between sugar
overconsumption and its behavioural correlates. Further, we attempted
to interpret this result and suggest possible neural mechanisms.
However, it should be noted that by proposing hypotheses about
changes in neural functioning we built on the previous extensive liter­
ature on the subject. We are aware that this approach does not exhaust
the subject and requires further research into neural processes and
changes involved in excessive sugar consumption.
CRediT authorship contribution statement
KM - Conceptualization; Funding acquisition; Methodology; Project
administration; Resources, Supervision; Validation; Writing – original
draft; Writing – review & editing; WP - Conceptualization; Data cura­
tion; Formal analysis; Methodology; Validation; Visualization; Writing –
original draft; Writing – review & editing; AC and KG - Conceptualiza­
tion; Investigation. All authors reviewed and accepted the manuscript.
Declaration of Competing Interest
The authors declare no competing interests.
Data availability
Data will be made available on request.
Acknowledgments
This research project was supported by the National Science Centre,
Poland – grant No. 2015/19/D/HS6/00781.

Appendix 1. Descriptive data for analysed variables

Trials

H T1 T2 T3

Diet Exp. Condition Mean Stdev Mean Stdev Mean Stdev Mean Stdev

Frequency of moving between the chamber zones

Standard ADD 20.400 2.187 17.733 2.890 20.733 2.865 17.200 3.707
MoveAdded 21.536 3.374 18.565 3.553 18.87 3.152 20.000 3.931
Sugar ADD 19.750 1.545 17.083 1.832 19.917 1.832 18.917 1.443
MopveAdded 18.556 2.185 17.667 2.309 19.75 2.527 20.833 2.406
Frequency of contact with the tunnels in the changed zone of the chamber
Standard ADD 7.178 1.068 8.600 2.131 8.267 2.052 8.600 1.724
MoveAdded 7.449 1.569 9.435 3.188 8.957 2.931 9.087 3.356
Sugar ADD 6.917 0.944 7.833 2.552 7.667 1.435 8.750 1.712
MoveAdded 7.444 2.066 8.250 1.658 7.667 2.934 9.750 3.223
Time spent on contact with the tunnels in the changed zone of the chamber
Standard ADD 64.795 13.608 140.413 32.392 125.827 22.969 156.588 33.762
MoveAdded 59.337 11.622 154.358 42.258 126.236 42.180 121.450 32.069
Sugar ADD 62.019 5.870 144.61 48.740 121.804 44.846 126.015 23.954
MoveAdded 62.307 9.412 130.749 26.083 92.479 24.701 99.255 26.859
Time spent in the changed zone of the chamber
Standard ADD 111.766 20.659 187.21 35.122 167.045 33.808 197.975 39.986
MoveAdded 102.868 20.377 194.264 41.981 163.389 34.944 163.793 38.668
Sugar ADD 104.241 11.622 182.926 50.334 160.503 26.949 179.519 36.307
MoveAdded 107.185 8.466 174.568 33.82 131.589 29.525 148.020 36.577
Time spent in the transporter
Standard ADD 73.150 6.676 55.018 27.64 54.177 25.573 52.716 24.202
MoveAdded 63.836 19.604 49.943 25.379 43.53 24.185 53.683 27.678
Sugar ADD 82.032 9.782 40.899 10.346 51.839 17.098 56.514 20.581
MoveAdded 80.382 16.921 52.996 20.152 56.511 13.592 63.932 20.596
Frequency of contact with the tunnels in the unchanged zone of the chamber
Standard ADD 7.511 0.958 6.267 1.387 6.733 1.280 5.600 1.682
MoveAdded 8.101 3.300 6.435 2.041 7.000 1.977 7.261 2.200
Sugar ADD 6.389 1.213 6.333 1.614 6.833 2.290 6.583 1.443
MoveAdded 6.694 1.291 6.667 1.670 8.333 1.723 6.917 1.621
Time spent on contact with the tunnels in the unchanged zone of the chamber
Standard ADD 60.774 8.279 49.524 15.589 57.449 17.947 50.298 14.215
MoveAdded 67.623 17.945 45.216 13.505 62.753 19.929 61.409 21.762
Sugar ADD 64.304 5.562 59.125 17.794 59.767 10.726 52.178 9.942
(continued on next page)

10
K. Modlinska et al. Behavioural Brain Research 436 (2023) 114085

(continued )
Trials

H T1 T2 T3

Diet Exp. Condition Mean Stdev Mean Stdev Mean Stdev Mean Stdev

MoveAdded 59.649 9.541 62.519 17.162 73.202 17.971 53.658 14.851

Time spent in the unchanged zone of the chamber
Standard ADD 105.452 7.018 81.414 17.742 94.524 25.874 78.136 14.166
MoveAdded 118.819 25.327 79.374 17.279 100.191 24.397 103.125 29.011
Sugar ADD 103.887 12.496 99.215 30.240 112.452 30.020 90.758 19.918
MoveAdded 99.685 13.345 98.27 26.794 125.517 29.801 93.716 25.249
Time spent in the central zone of the chamber
Standard ADD 133.02 11.135 98.601 25.95 103.692 26.823 88.886 34.661
MoveAdded 131.969 23.068 98.644 42.319 112.854 42.065 100.497 28.440
Sugar ADD 130.427 14.864 95.896 31.172 90.914 22.605 100.066 20.096
MoveAdded 130.826 20.128 90.48 28.395 104.578 37.489 116.273 26.472

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