Brain Struct Funct (2018) 223:897–912
https://doi.org/10.1007/s00429-017-1530-z
ORIGINAL ARTICLE
Inter-individual differences in decision-making, flexible and goal-
directed behaviors: novel insights within the prefronto-striatal
networks
Aurélie Fitoussi1,2 • Prisca Renault1,2 • Catherine Le Moine1,2 • Etienne Coutureau1,2
Martine Cador1,2 • Françoise Dellu-Hagedorn1,2,3
Received: 25 July 2015 / Accepted: 28 September 2017 / Published online: 12 October 2017
Ó Springer-Verlag GmbH Germany 2017
Abstract Inflexible behavior is a hallmark of several
decision-making-related disorders such as ADHD and
addiction. As in humans, a subset of healthy rats makes
poor decisions and prefers immediate larger rewards
despite suffering large losses in a rat gambling task (RGT).
They also display a combination of traits reminiscent of
addiction, notably inflexible behavior and perseverative
responses. The goal of the present work was twofold: (1) to
elucidate if behavioral inflexibility of poor decision-makers
could be related to a lower quality of goal-directed
behavior (action–outcome associations); (2) to uncover the
neural basis of inter-individual differences in goal-directed
behavior. We specifically assessed inter-individual differ-
ences in decision-making in the RGT, flexibility in the
RGT-reversed version and goal-directed behavior in a
contingency degradation test, i.e., response adaptation
when dissociating reward delivery from the animal’s
action. The contributions of the medial prefrontal cortex
and the dorsal striatum to action–outcome associations
were assessed using Zif268 immunodetection. Inflexible
behavior was related to a lower sensitivity to contingency
degradation in all poor decision-makers and only in a few
good decision-makers. This poorer sensitivity was associ-
ated with a lower immunoreactivity in prelimbic and
infralimbic cortices and a higher one in the dorsomedial
and dorsolateral striatum. These findings suggest that an
& Françoise Dellu-Hagedorn
francoise.dellu@u-bordeaux.fr
1
University of Bordeaux, INCIA, UMR 5287,
33000 Bordeaux, France
2
CNRS, INCIA, UMR 5287, 33000 Bordeaux, France
3
University of Bordeaux, INCIA, CNRS, UMR 5287, 146 rue
Léo Saignat, BP 31, 33076 Bordeaux Cedex, France
•
imbalanced prefronto-striatal activity could underlie inac-
curate goal representation in changing environments and
may promote maladaptive habit formation among poor
decision-makers. These data strengthen our previous work
identifying biomarkers of vulnerability to develop psychi-
atric disorders and demonstrate the relevance of inter-in-
dividual differences to model maladaptive behaviors.
Keywords Decision-making
Rat gambling task
Goal-
directed behavior
Inflexible behavior
Prefrontal cortex

Striatum
Introduction
Behavioral flexibility in a changing environment is fun-
damental in everyday life. Executive functions that allow
initiating, achieving and updating goal-directed actions
(Ernst et al. 2004; Fellows 2004; Lee et al. 2012) play a
pivotal role in flexible behaviors (Griffiths et al. 2014).
This role is exemplified by the deleterious consequences of
inflexible behavior as observed in several mental disorders
related to poor executive control (addiction, attention def-
icit-hyperactivity disorder, or obsessive–compulsive dis-
order) (Royall et al. 2002). These mental disorders are
typically characterized by poor decision-making, a deficit
that can be revealed in the Iowa gambling task (IGT) that
simulates complex and conflictual situations occurring in
real-life (Bechara et al. 1994). Importantly, poor decision-
making is also observed in a subset of healthy individuals,
for whom immediate small gratifications prevail over long-
term larger gain (Bechara and Damasio 2002; Bechara
et al. 2002).
Similarly, we identified in rodents a minority of poor
decision-makers in the rat gambling task (RGT) (Rivalan
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et al. 2009a), a task analogous to the human IGT,
demonstrating that rat behavior can reliably model
dimensions found in humans. The identification of extreme
or maladapted behaviors expressed spontaneously in rats
and comparable to those observed in the clinic has proven
to be useful to study the complexity of clinically relevant
phenotype and to discover endophenotypes, as no particu-
lar hypothesis is established about their origins (Bardo
et al. 1996; Belin and Deroche-Gamonet 2012; Blondeau
and Dellu-Hagedorn 2007; Deroche-Gamonet et al. 2004;
Kabbaj and Akil 2001; Koolhaas et al. 2007; LaPorte et al.
2010; Matzel and Kolata 2010; Rivalan et al. 2009b, 2013;
Robbins et al. 2012; Robinson et al. 2014). A thorough
appraisal of the poor decision-makers in our animal model
(i.e., behavioral, neurobiological phenotypes and their eti-
ology), could shed light on the pathogenesis of these
mental disorders and identify potential risk factors as they
may share common neuropsychological characteristics
with patients that could represent potential risk factors
(Rivalan et al. 2009b). In that line, we recently showed that
poor decision-making in rats can be accurately predicted
from a combination of behavioral and cognitive traits
reminiscent of psychiatric symptoms of decision-making-
related psychiatric disorders, namely risk-taking, reward-
seeking, motor impulsivity and behavioral inflexibility
(Rivalan et al. 2013). Behavioral inflexibility of poor
decision-makers was particularly noticeable in the RGT
reversal procedure, which requires redirecting choice on
the basis of new response-reward contingencies. Their
inflexible behavior was also associated with perseverative
and compulsive-like behaviors in different experimental
situations (Rivalan et al. 2013), suggesting an impairment
in goal-directed behavior for these individuals.
A plausible hypothesis is that this maladaptive behavior
could be related at least in part to an inefficient encoding
and updating of action–outcome (A–O) associations that
governs goal-directed behaviors. The quality of an A–O
association can be specifically assessed by degrading the
instrumental contingency, i.e., by dissociating reward
delivery from the animal’s action (i.e., lever press) (Bal-
leine and Dickinson 1998). If lever pressing is under the
control of the expected outcome (A–O), then that behavior
should progressively cease as the animal learns that lever
presses are no longer necessary to obtain rewards. Several
studies have demonstrated that the sensitivity of instru-
mental responses to contingency degradation requires the
integrity of a wide neuronal circuit which includes the
dorsal striatum (Yin et al. 2005, 2006), the mediodorsal
thalamus (Bradfield et al. 2013; Corbit et al. 2003), the
basolateral nucleus of the amygdala (Balleine et al. 2003),
and the medial prefrontal cortex (mPFC) (Corbit and Bal-
leine 2003; Coutureau et al. 2012; Naneix et al. 2009). At
present, however, it is not clear whether inter-individual
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Brain Struct Funct (2018) 223:897–912
differences in the quality of the encoding of A–O associ-
ations are related to differences in these network activities.
The aim of the present work was twofold. First, to relate
inter-individual differences in decision-making to basic
cognitive processes, we assessed, within the same sample
of individuals, decision-making performances in the RGT,
behavioral flexibility in the RGT reversal procedure and
sensitivity to contingency degradation in instrumental
conditioning. Second, we aimed to uncover the neural basis
of inter-individual differences in the quality of A–O asso-
ciations through the expression of the immediate early gene
Zif268 within prefronto-striatal networks. Zif268 was
chosen here since it is a transcription factor well charac-
terized for its role in synaptic plasticity and memory pro-
cessing (Bozon et al. 2003) and has been shown to be
crucial in instrumental learning (Maroteaux et al. 2014).
Materials and methods
Animals
Thirty male Wistar Han rats (Charles River, Lyon, France)
aged from 13 to 15 weeks were used. They were housed in
groups of four in a temperature-controlled room (22 °C) on
an inverted 12-h light/dark cycle (light on at 8:00 PM).
Tests were conducted during the dark phase of the cycle.
The rats had free access to water and were moderately food
deprived (95% of free feeding weight) throughout the
experiments. All procedures were conducted in strict
accordance with the 2010-63-EU and with approval of the
Bordeaux University Animal Care and Use Committee
(Permit number: 5012087-A).
Behavioral apparatus and procedures
Experimental design and principle of the RGT and RGT-
reversed version are given in Fig. 1a.
Four rats were exposed to the same behavioral tests as
the others except for the contingency degradation test
during which they served as control rats (CONT = non-
degraded condition). To compare the scores of exactly the
same rats through all the experiments, scores of these
control rats were excluded from the data analysis of the
behavioral tests.
Decision-making in the rat gambling task
Principle The RGT requires the rat to deduce, by trial and
error, among four options, the two that are the more
rewarding on the long-term and tracks the continuous and
dynamic process of deduction and readjustment of choice
(de Visser et al. 2011; Rivalan et al. 2009a). The principle
Brain Struct Funct (2018) 223:897–912 899

a RGT RGT-reversed version

choice reward penalty
food
dispenser
D C D very short (1/2) D C
C short (1/4)
B B
A B very long (1/4)
A long (1/2)
A
holes for choices
Choice of an advantageous opon Choice of an advantageous opon

Operant training RGT RGT-rev

5d 1d 5d 1d

b Conngency degradaon paradigm

Operant training Conngency degradaon test

retraining
Operant training INFLEX-I
CD test1 RR10 CD test2
FLEX-S
FR1 RR5 RR10 1 month 6d 3d +60 min
RR10 CONT
2d 3d 5d
6d RR10 RR10
1d 1d

Fig. 1 Experimental designs and principles of the behavioral tasks: a
the RGT and RGT-reversed version and b the contingency degrada-
tion task. In the RGT, choices A and B allowed delivery of two pellets
immediately, but were followed by long and unpredictable penalties
(time-out). By contrast, C and D choices allowed the delivery of only
one pellet immediately but were followed by shorter penalties. The
time-out could occur according to a low probability (1/4) for B and C
choices or a high probability (1/2) for A and D. Thus, A and B choices
were equally disadvantageous whereas C and D were equally
advantageous. Advantageous choices could provide up to fivefold
more pellets than disadvantageous choices. In the RGT-reversed
version (RGT-rev), A–B and C–D associated outcomes were spatially
reversed. b In the contingency degradation paradigm, rats were first
trained under a fixed-ratio (FR1) and random ratio (RR5 and 10)
schedules of reinforcement. Contingency degradation (CD) test
consisted in delivering pellets independently of lever presses. Rats
were killed 60 min after the end of the second test for Zif
immunoreactivity quantification and comparisons between rats sen-
sitive to contingency degradation (FLEX-S) and those less sensitive
(INFLEX-I). They were compared to control rats (CONT) that only
had 2 RR10 sessions, a week apart

of the task tightly mimics that of the human IGT: two
options (holes chosen by nose-poking) that steadily offer
bigger immediate food reward, are disadvantageous in the
long run due to higher unpredictable penalties (frustrating
time-outs occurring after food delivery, during which no
reward can be obtained). Conversely, the two advantageous
options steadily offer smaller reward, but unpre-
dictable penalties that can follow are shorter. A choice also
results in the deactivation of all stimulus-lights except for
the chosen hole, until the reward is collected or the penalty,
if it occurs, ends up. This is particularly important during
the time-out periods to facilitate the association between
hole-response and its consequences. Rats are free to make a
new choice whenever the reward has been collected or
whenever the penalty finishes (no inter-trial interval).
Apparatus The experiments were performed in twelve
polyvalent conditioning boxes (Imetronic, Pessac, France;
28 9 30 9 34 cm). Boxes were equipped with four nose-
poke holes, dimly illuminated within the hole with a white
LED. These holes were located on a curved wall on one
side of the box, equidistant to a food magazine situated on
the opposite wall. Each hole was equipped with an infrared

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detector connected to an external dispenser delivering food
pellets (45 mg, formula P, Sandow scientific, USA). Data
collection was made using a control software (Imetronic,
Pessac, France) running on a computer outside the testing
room. At least 30 min before each session, the rats were
placed in the light-attenuated and temperature-controlled
(23 °C) experimental room.
Training Procedures were previously described (Rivalan
et al. 2009a, 2013). The training phase consisted in learn-
ing to associate two consecutive nose-pokes in one of the
four illuminated holes with the delivery of one or two food
pellets in the magazine. For this purpose, rats had first to
associate a single nose-poke in any of the four illuminated
holes with the delivery of one food pellet in the magazine.
After a nose poke, only the selected hole remained illu-
minated, but all were inactivated until the rat collected the
food reward. After food collection, all the holes were
lightened and activated again. This procedure continued
daily until rats obtained 100 pellets within a session
(30 min cut-off). Then two consecutive nose pokes in the
same hole were required to obtain food, to ensure that the
selection of the hole was a voluntary choice. After reaching
the same criterion, rats were submitted to two final 15 min
training sessions. In the first session, two pellets were
delivered after a choice was made (maximum 50 pellets).
Then, within a second session, one pellet is delivered at a
time (maximum 50 pellets). These sessions habituated the
rats to the quantity of pellets that could be obtained during
the test. The training phase usually lasted 5–7 days and
tests were performed the following day.
Test Rats could freely choose between four nose-poke
holes (A–B–C–D) during a 1-h test session (or max. 250
pellets obtained). Choices C and D vs A and B led to the
immediate delivery of one vs two pellets, but A and B
choices could be followed by longer, unpre-
dictable penalties (222 and 444 s time-outs) compared to C
and D choices (12 and 6 s). Penalties occurred at a low
probability (1/4) for B and C choices, and at a high prob-
ability (1/2) for choices A and D. A time-out could not
occur before the third or fourth choice, to favor sampling of
the different conditions at the beginning of the test. During
the penalty, all lights were switched off and nose-poke
holes were disabled, but the chosen hole remained illumi-
nated to facilitate association between each choice and its
consequences. A brief extinction of this light (1 s) signaled
the end of the time-out. The theoretical maximum gain was
the same for advantageous C and D choices, and five times
higher than for disadvantageous A and B choices. We used
the same criteria as in our previous works with this task, to
select animals according to their performance (Fitoussi
et al. 2015; Rivalan et al. 2009a, 2013). Good (GD) and
poor (PD) decision-makers were differentiated on the basis
of the percentage of advantageous choices ([ 70 and
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Brain Struct Funct (2018) 223:897–912
\ 30% respectively) during the last 20 min of test. The
remaining rats with intermediate scores were classified as
undecided (between 30 and 70% of advantageous choices).
These rats (n = 4) were not further studied because they
are too few in this experiment and overall only represent
16% of the population as shown in a previously published
meta-analysis (Rivalan et al. 2013). Only rats that sampled
both advantageous and disadvantageous options during the
first 20 min of the test were considered. The mean latency
to collect food pellets was recorded as a motivational food
reward index.
Behavioral flexibility in the rat gambling task-reversed
version
In a second stage, 5 days later, A–B and C–D associated
outcomes were spatially reversed to assess behavioral
flexibility (Rivalan et al. 2013). The test again consisted in
a single 60 min test session. Distinct behaviors can be
observed during reversal, as previously shown (Rivalan
et al. 2013), that are very distinct from behaviors in the
RGT. For example, a rat can gradually reverse its choices
towards the new location of its favorite options (flexible
rat) or a rat can remain during all the session on the options
that it preferred during test (inflexible rat). An index of
flexibility was calculated as the mean percentage for the
options preferred during the RGT, whether advantageous
or disadvantageous. Behaviors were differentiated on the
basis of the time-course of choices and flexibility and were
classified into two categories: flexible with gradual rever-
sion towards the new location of their favorite options
([ 60% of reversed chosen options during the last 20 min)
and inflexible with perseveration of previously learned
choices (\ 40% of choices). The use of a different criteria
to distinguish between flexible and inflexible behaviors
(see Rivalan et al. 2013), is justified by the nature of the
task which is radically different (learning vs reversal). Rats
with flexible behavior cannot reach a high level of pref-
erence as fast as in the RGT, because of the difficulty of
reversing choices.
Goal-directed behavior during contingency degradation
Experimental design and principle of the contingency
degradation procedures are given in Fig. 1b. Two contin-
gency degradation experiments were made with the same
rats, a month apart. The first was used to describe inter-
individual differences in performance, notably the sessions
necessary to reveal the largest variations in behavior. The
second was a control of the reliability of the measure and
was used for Zif268 immunodetection. The best condition
for this assessment (third session) was chosen according to
the previous experiment and to the performances obtained
Brain Struct Funct (2018) 223:897–912
during this test. The control group (CONT) was made by
pooling two undecided and two good decision-makers and
was tested under un-degraded condition.
Apparatus Animals were trained in eight identical con-
ditioning chambers (32 cm wide 9 32 cm deep 9 22 cm
high, Imetronic), each located inside a sound and light-
attenuating chamber. Boxes were constructed from white
plastic panels with a Plexiglas door and were equipped
with a fan providing a background sound. Each box was
permanently illuminated by a diffuse 2 lux light source
located at the middle of the ceiling. The floor consisted of
stainless steel bars 5 mm in diameter spaced 1.5 cm apart.
One stainless steel lever protruded horizontally 1 cm from
the wall situated at the left of the door, 6 cm above the grid
floor. A tray was situated centrally on the opposite wall.
Food pellets (45 mg, Formula P, Sandow scientific, USA)
were delivered in the magazine from a food dispenser. The
magazine was equipped with infrared cells to detect the
animal’s visits. A program (Imetronic, Pessac, France)
controlled the chambers and collected the data on a
microcomputer (outside the testing room).
Instrumental training Rats were trained to press a lever
to obtain a reward during daily training sessions. Different
reinforcement schedules were used. The rats first received
training under a continuous reinforcement, fixed ratio (FR)
1 schedule, for 2 days (each lever press was rewarded).
Animals were then shifted to a random ratio (RR) 5
schedule for 3 days; each response was rewarded with a
probability of 0.2 on average, meaning five lever presses
rewarded on average) and then to a random ratio 10
schedule for 4 days (RR-10, each response was rewarded
with a probability of 0.1 on average, meaning 10 lever
presses rewarded on average). Each session ended when
rats had earned 100 pellets or when 30 min had elapsed.
Contingency degradation The test consisted in six daily
consecutive sessions. Animals received response-indepen-
dent pellet delivery on a random time schedule so that the
instrumental contingency was degraded: pellet deliveries
occurred independently of lever presses. Response-inde-
pendent reward deliveries were yoked to reward deliveries
earned during the last RR-10 session for each rat (see
Table 1) and was used as a reference for the individual
delivery program, independently of animal lever presses,
during the testing days. This procedure reduces the influ-
ence of variation in levels of activity and results in a
similar frequency of reward delivery during the transition
between training and test. For the CONT (undecided and
good decision makers), the A–O contingency was
unchanged. Each session ended when rats had received 100
pellets or when 30 min had elapsed. Behavioral data were
displayed as the percentage of lever presses during test
compared to the last training session. An index of general
901
activity was measured by the sum of lever presses and
visits to the magazine per min.
Individual sensitivity to contingency degradation
Training One month later, rats were trained again under the
RR-10 training schedule during six consecutive sessions
for the experimental group, and only one session for the
control group (CONT) so that lever response rate could be
comparable between groups during testing days.
Test Rats were given a second contingency degradation
test, similar as the one previously used. Animals were
tested during three consecutive days, except the CONT
group which was run on the corresponding first and last
testing days. Given the results of the first experiment, we
expected that the third session would be appropriate to
ensure large inter-individual differences. After the last
session, animals were moved to a quiet room and killed
60 min later.
Zif immunohistochemistry and cell counting
Because this study is based on inter-individual differences,
we had to reduce, whenever possible, any possible source
of variation in Zif268 quantification. After the last behav-
ioral testing (30 min), rats had to be in quiet conditions
(1 h apart in a quiet room) before being killed for Zif268
analyses. We also had to organize the experiment in a way
to minimize variability related to time-schedule (morning
sessions) and to housing (4 animals per cage): four rats
were tested at a time (1 home cage), then, placed in their
home cage in a quiet room for 1 h and then, we proceeded
to four perfusions at the same time. The following groups
were tested later, with a delay allowing the best technical
conditions. For these reasons, only 12 rats per day for
2 days were used for Zif268 quantification (n = 24).
Rats were deeply anesthetized with sodium pentobar-
bital (Céva Santé Animale, France) and perfused through
the heart with 200 ml of NaCl 0.9%, followed by 250 ml of
paraformaldehyde (PFA) 4% in 0.1 M phosphate buffer
(PB). Brains were dissected out, post-fixed 24 h in PFA 4%
and transferred into a PB 0.1 M/sucrose 30% solution for
48 h at 4 °C. Brains were frozen and stored at -80 °C until
use. Serial coronal sections (50 lm) were cut on a freezing
microtome (Leica SM 2400), collected, and stored in a
cryoprotectant solution.
After the initial rinsing (3 9 10 min in PBS), all the
steps of the immunohistochemistry procedure were sepa-
rated by 3 9 10 min rinsing step with PBS (0.1 M pH 7.4)
under agitation. A rabbit affinity purified polyclonal anti-
Zif268 antibody (Sc-189, Santa-Cruz Biotechnology Inc,
CA, USA) was used as a primary antibody (1:3000 in 0.3%
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Table 1 Parameters of the

RGT FLEX Rat Random time schedule (RT) (s) Mean RT (s)
random time schedule during
contingency degradation test. PD INFLEX 3 10 \ 12 \ 15 12
Random time schedules (RT) in
seconds and mean RT are PD INFLEX 9 24 \ 30 \ 35 30
calculated for each rat, to be PD INFLEX 17 12 \ 15 \ 18 15
yoked to reward deliveries PD INFLEX 20 15 \ 19 \ 23 19
during the last random-ratio 10
PD INFLEX 25 12 \ 15 \ 18 15
session
PD INFLEX 26 12 \ 15 \ 18 15
PD INFLEX 30 12 \ 15 \ 18 15
GD INFLEX 11 12 \ 15 \ 18 15
GD INFLEX 19 12 \ 15 \ 18 15
GD INFLEX 21 24 \ 30 \ 35 30
GD INFLEX 23 10 \ 12 \ 15 12
GD INFLEX 28 12 \ 15 \ 18 15
GD FLEX 1 12 \ 15 \ 18 15
GD FLEX 5 12 \ 15 \ 18 15
GD FLEX 13 12 \ 15 \ 18 15
GD FLEX 14 15 \ 19 \ 23 19
GD FLEX 16 15 \ 19 \ 23 19
GD FLEX 18 12 \ 15 \ 18 15
GD FLEX 22 24 \ 30 \ 35 30
GD FLEX 24 12 \ 15 \ 18 15
GD FLEX 4 15 \ 19 \ 23 19
GD FLEX 6 15 \ 19 \ 23 19
GD FLEX 8 10 \ 12 \ 15 12
GD FLEX 12 10 \ 12 \ 15 12
GD FLEX 15 12 \ 15 \ 18 15
Random time schedules (RT) in seconds and mean RT calculated for each rat, for contingency degradation
test

PBS-triton/3% normal donkey serum). Free-floating sec-
tions were incubated with the primary anti-Zif268 antibody
overnight at room temperature (20 °C) under agitation.
After rinsing, sections were then incubated with a
biotinylated Donkey anti-rabbit secondary antibody
(1:2000 in PBS-triton 0.3%, GE Healthcare, UK) for 2 h at
room temperature, followed by the avidin–biotin–peroxi-
dase complex (1:800 in PBS, Vector Laboratories) for 2 h
at room temperature. The final staining was obtained fol-
lowing incubation with diaminobenzidine (DAB, Sigma-
Aldrich 0.2 mg/ml)-Nickel (0.04 mg/ml) in Tris-buffered
saline (pH 7.6) containing 0.003% H2O2 (Sigma). Sec-
tions were finally rinsed three times with 0.1 M Tris buffer,
disposed onto gelatin-coated slides, and mounted in Eukitt
after 24 h of air-drying.
For quantification, neurons with nuclei exhibiting
Zif268 immunolabeling were counted bilaterally in sec-
tions taken through prefrontal and striatal areas according
to the rat atlas of Paxinos and Watson (1997). Details of
sub-regions and anatomical coordinates (indicated from
Bregma) were chosen as follow: the prelimbic cortex (PL;
? 3.20 mm), the infralimbic cortex (IL; ? 3.20 mm), the
anterior cingulate cortex (CgA; ? 1.60 mm), the anterior
and posterior parts of the dorsomedial striatum (DMSa and
DMSp, respectively; ? 1.60/0.20 mm) and the anterior and
posterior part of the dorsolateral striatum (DLSa and
DLSp; ? 1.60/0.20 mm). Anterior and posterior striatal
areas were distinguished since functional differences
between these subdivisions were previously described (Yin
et al. 2005). For each area, counting was performed within
the entire region in three consecutive sections per rat by an
experimenter blind to the experimental groups. Zif268-
positive neurons were quantified using the Mercator soft-
ware (Explora Nova, La Rochelle, France). Results were
expressed as number of zif268-positive nuclei per
mm2 ± SEM.
General data analysis
In both the RGT and the RGT-reversed version, behavioral
scores, i.e., percentage of advantageous choices
(mean ± SEM) were compared to chance (50%) using a

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two-tailed one-sample t test for subgroups (Good and Poor
decision-makers). Comparisons of proportions of individ-
uals with analogous preference during training and RGT
were conducted using the non-parametric Fisher’s exact
test (StatXact 9). Two-way ANOVAs were used to com-
pare groups [repeated measures (RM) for time or structure
and group factors] followed by simple main effects (SME)
or post hoc Newman–Keuls (NK) tests, when required
(Statistica, Statsoft 7.1). Controls of normality distribution
and homogeneity of variance were made. Comparisons of
the number of Zif268-positive nuclei between groups were
made for subcortical and cortical areas separately. Corre-
lations between behavioral scores and region-specific
immunoreactivity levels were made using the parametric
Bravais-Pearson’s correlation test (Statistica, Statsoft 7.1).
Results
All poor decision-makers and only some good
decision-makers demonstrate behavioral inflexibility
Inter-individual differences in decision-making
during the RGT
The RGT measures, across successive trials, the ability to
make the most advantageous choices. In this task, choices
associated with a higher immediate gain are disadvanta-
geous in the long run due to higher unpredictable penalties.
As previously shown, we found distinct patterns of choice
preference within a single RGT session (Fig. 2a). A ran-
dom selection of the options was necessary to avoid a
preference bias in choices. All rats initially selected both
kinds of options randomly before developing a preference
except one that did not chose all the options and was dis-
carded. Rats (n = 25) clustered into two main categories
depending on their final preference (Fig. 2b). A majority of
good decision-makers (n = 18, 64%) that displayed a
strong preference for advantageous options ([ 70% pref-
erence) and a minority of poor decision-makers (n = 7,
25%) that chose disadvantageous options ([ 70%
preference).
We first verified if choices during the RGT were unre-
lated to a side preference eventually developed during
training. Thus, for each individual, we compared the
options preferred during the last day of training and those
preferred during test. A preference was noticed when the
options were chosen more than 70% of the total number of
choices (spatially nearby A and B: AB or spatially nearby
C and D: CD). No preference was noted when options AB
and CD were chosen less than 70% (ABCD). Given that
three kinds of preference could be made (preference for
options CD, for options AB or no preference ABCD),
903
chance level to choose the same options is 1/3. Nine rats
out of 25 had similar preference (36%), a proportion that
does not differ from 33% (Fisher exact test, p = 1; ns).
Thus, choices made during the test were not influenced by
those eventually developed during training. Good and poor
decision-makers also differed on their within-session pat-
tern of choices. Whereas good decision-makers tended (on
average) to develop progressively their preference toward
the best options, all poor decision-makers quickly dis-
played a preference for disadvantageous options, probably
because of a preference for the higher amount of immediate
reward and lack of penalty during initial sampling. Con-
sequently, pellet consumption (Fig. 2c) differed greatly
between groups (F1,23 = 15.23, p \ 0.001) and was much
lower for poor decision-makers over time (interaction
group 9 sessions, F5,115 = 17.53, p \ 0.001). As previ-
ously shown, poor decision makers also displayed a shorter
latency to collect food, that we consider as a higher
motivation for the reward (t = 2.4, df = 22, p \ 0.05)
(Fig. 2d).
Behavioral flexibility during the RGT-reversed version
Reversing contingencies in the RGT measures subjects’
behavioral flexibility in shifting their behavior when
advantageous and disadvantageous choices were spatially
reversed. Persistence to choose the same location reveals
behavioral inflexibility (INFLEX), whereas changes in
choices reflect detection of the change and behavioral
flexibility. Detection of the change was demonstrated by
undecided behavior and then reversed choices (FLEX).
100% of poor (n = 7) vs only 28% (n = 5) of good
decision makers were INFLEX and thus, chose the same
hole location (i.e., inflexible behavior) (Fig. 2e, f). Most
good decision-makers (72%, n = 13) were FLEX. In a
previous study (Rivalan et al. 2013), we obtained very
similar results that corroborate the present findings (all
poor decision-makers (100%; 6/6) vs only a third (36%;
5/14) of good decision-makers were inflexible).
Behavioral inflexibility is associated with a lower
sensitivity to the contingency degradation procedure
Operant training
The four groups of animals (INFLEX poor and good
decision-makers, FLEX good decision-makers and CONT)
acquired the instrumental response (operant training phase)
at the same rate (Fig. 3a). A mixed ANOVA with session
and group as factors showed an effect of session
(F9,243 = 39.23; p \ 0.001), but no effect of group
(F3,27 = 0.65; ns) nor any significant group 9 session
interaction (F27,243 = 0.46; ns), indicating that all groups
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Fig. 2 Inter-individual differences during the RGT and RGT-re-
versed version. a Time-course of advantageous choices (%) during
the rat gambling task of good decision-makers (GD; n = 18) and poor
decision-makers (PD; n = 7); b individual RGT score during the last
20 min for GD and PD; c cumulated number of pellets earned during
the task; d mean latency to collect food during the RGT; e time-
course of advantageous choices during the RGT-reversed version of
PD and GD. Inflexible rats displayed less than 40% of reversed option
similarly acquired the instrumental response. Mean
response rates of INFLEX poor and good decision-makers,
FLEX good decision-makers and CONT at the completion
of training were 38.1, 38.4, 39.7, and 40.1, respectively.
Contingency degradation
Figure 3b shows the relative rate of responses (i.e., fre-
quency of lever presses during the testing sessions as
compared to the last training session) for all the groups,
with marked differences. No significant difference between
groups was observed in the mean RT applied [INFLEX PD:
17.3 ± 2.25; INLEX GD: 17.4 ± 3.2; FLEX GD:
16.9 ± 1.29; F(2,22) = 0.017, ns]. The FLEX group dis-
played an important decrease in lever pressing, indicating
that these animals correctly adapted their operant responses
to the changes in action–outcome contingency. Interest-
ingly, the two INFLEX groups had a much slower decrease
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Brain Struct Funct (2018) 223:897–912
preferences (INFLEX PD or GD), flexible rats that reversed their
choices (FLEX) displayed more than 60% during the last 20 min. f
Proportions of GD and PD according to their performance in the
RGT-reversed version. Dotted line represents chance level (50%).
Comparisons with chance level, Student’s t test (a, e); comparisons
between groups (c, d) NK post hoc analysis and Student’s t test,
respectively, #p \ 0.05, *p \ 0.01
in lever pressing (mean of 51.0 and 48.9, respectively on
the 6th day of training, compared to 23.3 for FLEX),
indicating that these groups had an impairment in adapting
their behavioral responses to action–outcome contingency
changes. The ANOVA with Session and degraded Groups
as factors confirms the dissociation between INFLEX and
FLEX. It reveals a significant effect of session
(F5,110 = 41.24, p \ 0.001) and group (F2,22 = 11.52,
p \ 0.001) and a significant interaction between session
and group, (F10,110 = 2.72, p \ 0.01). Specifically, the
INFLEX groups did not differ from each other (SME;
F1,22 = 0.36, ns). These INFLEX groups significantly
differed from FLEX group during testing days (INFLEX
poor decision makers vs FLEX; SME; F1,22 = 14.09,
p \ 0.001; INFLEX good decision makers vs FLEX;
F1,22 = 16.11, p \ 0.001). On the other hand, the level of
presses remained high and stable during the whole testing
for rats run in the non-degraded schedule [Comparisons
Brain Struct Funct (2018) 223:897–912
Fig. 3 Individual-based contingency degradation test. a Rate of lever
presses per min during training sessions for good decision-makers
(GD) with flexible reversed choices (FLEX) and with inflexible
choices (INFLEX GD and INFLEX PD); b relative responses rate
(ratio of lever press frequencies during the testing sessions and during
between sessions: Newman–Keuls (NK), ns]. Finally,
flexibility in the RGT-reversed version was inversely cor-
related with the sensitivity to the degradation contingency
procedure (index of flexibility vs relative response rate
during the fourth testing day: r = - 0.59; n = 24;
p \ 0.01).
Figure 3c shows the frequency of visits to the empty
food magazine. The more the animals learn that food pel-
lets are delivered independently of their action, the more
frequently they will check the food magazine. An ANOVA
comparing the rate of visits to the magazine between
groups confirms these behavioral differences (F3,27 = 4.05,
p \ 0.02) and reveals differences between groups within
the testing session (ANOVA group 9 sessions,
F15,135 = 2.06, p \ 0.02). FLEX group significantly
increased its visits to empty magazine per min across
sessions (F1,5 = 8.74; p \ 0.001). This increase was sig-
nificantly compared to the first session as early as the 3rd
session (NK, p \ 0.05).
Sensitivity to contingency degradation
and prefronto-striatal circuits
Here, we were interested in identifying the neural corre-
lates of inter-individual differences in the sensitivity to
the contingency degradation procedure by the measure of
Zif268 expression. For this purpose, rats were re-tested in
the contingency degradation procedure. Twenty-four rats
were used for Zif268 immunodetection during the con-
tingency degradation test: 10 FLEX rats, 10 INFLEX rats
and 4 CONT. Zif268 expression analyses of four rats
could not be exploited for technical reasons (absence of
905
the last training session) during the individual-based contingency
degradation test; and c frequency of visits to the empty magazine per
min. Comparisons between FLEX and INFLEX groups, NK post hoc
analysis, **p \ 0.01; *p \ 0.05; #p \ 0.05
correct labeling for reliable quantification). To study the
neurofunctional bases of inter-individual differences in
sensitivity to contingency degradation, rats were split into
two groups depending on their sensitivity to the procedure
during the 3rd day of testing, when brains were removed
for Zif268 immunochemistry processing. The criterion
chosen was the median score of relative response rate
(44%). Thus, one group (n = 8) included rats sensitive
(S) to contingency degradation with a score lower than the
median score. Because all these rats were also flexible in
the RGT-reversed task, they were named the FLEX-S
group. The other group (n = 8) included rats with a lower
sensitivity (insensitive, I) to contingency degradation,
with a score higher than the median. Most of these rats
were also inflexible in the RGT-reversed task and were
named the INFLEX-I group. The CONT group (n = 4)
was used for the non-degraded condition during testing
sessions 1 and 3.
Operant re-training
The previous contingency degradation procedure facili-
tated training since both groups reached much sooner (third
session) the same level of lever presses as at the end of the
first training phase. A mixed ANOVA with session and
group as factors showed an effect of session
(F4,56 = 11.05, p \ 0.001), but no effect of group
(F1,14 = 0.15, ns) nor any significant group 9 session
interaction (F4,56 = 0.71, ns), indicating that both groups
reacquired similarly the instrumental response during the
operant training.
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Contingency degradation before Zif268 analysis
Again, large inter-individual differences in sensitivity to
the contingency degradation procedure were observed
during the test sessions, notably during the third session
where FLEX-S and INFLEX-I displayed very different
performances. Figure 4a shows the effect of the contin-
gency degradation procedure on instrumental responses for
FLEX-S and INFLEX-I during the three testing days. As
expected, a marked difference distinguished FLEX-S and
INFLEX-I for lever pressing during all testing days [a
mixed ANOVA with days and group as factors showed an
effect of group (F1,14 = 17.08, p \ 0.001)]. Unlike the first
round of experiment, we report here a significant difference
between groups during the first day (NK, p \ 0.02), which
persisted during the following days (NK, p \ 0.01).
Whereas INFLEX-I did not decrease their response rate
compared to training during the first testing day (93.6%;
t = 0.87, df = 7, ns), the response rate of FLEX-S was
decreased by almost half (53.5%; t = 3.63, df = 7,
p \ 0.01). Then, the decrease in response rate followed a
similar time-course for the two groups (interaction
group 9 session, (F2,28 = 0.68, ns). During the 3rd day of
testing (testing day of interest for the analysis of Zif
expression), the response rate of INFLEX-I was only
decreased by 28%, whereas it was reduced by about 79% in
FLEX-S. Consequently, the response rate of the control
group in the non-degraded condition that remained
stable compared to training (100.2%), was much higher
Fig. 4 Individual-based contingency degradation test: second mea-
sure to assess neural markers of inter-individual differences. a
Relative response rates of flexible rats, sensitive to contingency
degradation (FLEX-S) and inflexible rats rather insensitive (INFLEX-
I) and control (CONT) groups during the individual-based contin-
gency degradation test; and b frequency of visits to the empty
magazine in number per min of FLEX-S and INFLEX-I and CONT
groups. For CONT group, values of the 1st session (training) and the
2nd session (test) are given. Comparisons between groups, NK post
hoc analysis, **p \ 0.01; *p \ 0.05
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Brain Struct Funct (2018) 223:897–912
than the FLEX-S under the degraded condition, but did not
significantly differ from INFLEX-I (F2,18 = 14.8
p \ 0.001; CONT vs FLEX-S, NK, p \ 0.001; CONT vs
INFLEX-I p = 0.07). An ANOVA comparing the rate of
magazine visits between the two groups during the three
testing days similarly showed marked differences
(F1,14 = 13.68, p \ 0.01) (Fig. 4b). During the 3rd day of
testing, the number of visits to the empty magazine of
CONT was similar to that of INFLEX-I (effect of CONT,
FLEX-S INFLEX-I groups, F2,18 = 6.18, p \ 0.01; NK,
ns) but was lower than that of FLEX-S (NK, p \ 0.02),
indicating that these animals have learnt that food pellets
are delivered independently of their action, since they
check more frequently for food in the magazine.
Region-specific differences in Zif268 positive-nuclei
within the prefronto-striatal circuits
Our data emphasize specific changes in Zif268 expression
related to the sensitivity to the contingency degradation
(last session) within the mPFC and dorsal striatal areas. We
focused on these specific networks since pilot works did
not show any difference in other brain regions. Specifi-
cally, animals which exhibited low Zif268 expression in
the PFC expressed a high expression in striatal regions and
reciprocally, suggesting that activities in these areas were
inversely related. Figure 5a, b illustrates the pattern of
Zif268 expression for FLEX-S and INFLEX-I groups.
Among brain areas, we report differences in Zif268
expression in two areas of the mPFC: the infralimbic cortex
(IL) and the prelimbic cortex (PL) as well as in the dorsal
striatum. FLEX-S rats displayed a higher density of
Zif268-positive nuclei than INFLEX-I in IL and PL (Two-
way RM-ANOVA, F1,14 = 6.30; p \ 0.05 for group fac-
tor; post hoc Fisher’s LSD, PL and IL, p \ 0.05) and a
lower number in dorsal striatal areas, notably in DLSa,
DLSp and DMSp (Two-way RM-ANOVA, F1,14 = 6.64,
p \ 0.05 for group factor; post hoc Fisher’s LSD,
p \ 0.05). Despite a clear tendency, we did not report any
significant difference in the cingulate cortex and DMSa
(post hoc Fisher’s LSD, ns).
It is noteworthy that significant correlations between level
of Zif268-positive neurons and behavioral sensitivity to
contingency degradation were reported. Negative correla-
tions were reported between the mPFC (both PL:
r = - 0.49, n = 16, p = 0.05 and IL: r = - 0.55, n = 16,
p \ 0.05) and relative response rate during contingency
degradation on the third session. Conversely, in the DLSa
and DMSp, we observed a positive correlation between the
density of positive-Zif268 nuclei and behavioral data
(r = 0.64, n = 16, p \ 0.01 and r = 0.55, n = 16,
p \ 0.05, respectively). Interestingly, these opposite trends
were also illustrated by significant negative correlations
Brain Struct Funct (2018) 223:897–912
Fig. 5 Quantification of Zif268 immunoreactivity in a the medial
prefrontal cortex and b the dorsal striatum in FLEX-S and INFLEX-I
groups. Results are expressed as the mean ± SEM of the number of
Zif268-positive neurons per mm2. Statistical differences between
groups, post hoc Fisher’s LSD analysis: *p \ 0.02. Dotted lines
represent the mean level of 4 control rats. Immunoreactivity
correlations with sensitivity to contingency degradation during the
third session are shown for the ratios between c the Prelimbic area and
between mPFC and striatal Zif268 ratios and behavioral
sensitivity to contingency degradation (see Fig. 5c–h),
whereas no significant correlation was found between any of
these ratios and the index of activity (index of activity vs
ratios PL/DLSa and DLSp: r = - 0.25 and - 0.33; n = 20,
907
the Dorsolateral striatum anterior part, d the Prelimbic area and the
Dorsolateral striatum posterior part; e the infralimbic area and the
dorsolateral part of the striatum, anterior part and f the infralimbic
area and the dorsomedian striatum posterior part. PL prelimbic cortex,
IL infralimbic cortex, CgA anterior cingulate cortex, DMS dorsome-
dian striatum, DMSa anterior part, DMSp posterior part, DLS
dorsolateral striatum, DLSa anterior part, DLSp posterior part
ns; vs ratios IL/DLSa and DSMp: r = 0.42 and - 0.29,
n = 20, ns). A positive correlation was observed between the
density of positive-Zif268 nuclei in DLSa and DMSa for
FLEX-S rats (r = 0.72; df = 7, p \ 0.05), whereas it was
not significant for INFLEX-I rats (r = 0.23, df = 7, ns).
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Discussion
The current results suggest that inflexible behavior and
poor decision-making in the RGT may be linked to a
failure in updating the A–O association that mediates goal-
directed behavior as measured during the contingency
degradation paradigm. This disability appears associated
with a specific imbalanced activity within the prefronto-
striatal networks.
How behavioral inflexibility may influence decision-
making
Like most of the currently available RGTs, our task inte-
grates multiple cognitive abilities involved in executive
functioning (de Visser et al. 2011; Fitoussi et al. 2015;
Rivalan et al. 2013; van den Bos et al. 2014; Zeeb et al.
2009). Additionally, we show that this complex task
reveals poor versus good decision-makers within a healthy
population, these behaviors being highly reproducible and
stable across time (Fitoussi et al. 2015; Rivalan et al.
2009a, 2011, 2013). We previously showed that poor
decision-making in the RGT is not related to a slower
learning, since one session is long enough for the behavior
to stabilize and to reveal option preference. Indeed, when a
second test is performed the following day, good and poor
decision-makers maintain the same preference developed
during the first test, throughout the entire session (Fitoussi
et al. 2015). Moreover, poor decision-making persisted
despite prior knowledge about the option values, learnt
separately in a task variant, indicating that they did not
choose the disadvantageous options because they failed to
acquire relevant information about the task (Fig. 5e; Riv-
alan et al. 2009a).
As previously shown (Rivalan et al. 2009a), choices
during the RGT was unrelated to a side preference even-
tually developed during training since no bias effect of
location preference eventually developed during training
was observed: the proportion of rats with identical prefer-
ence during training and test did not differ from chance.
Uncontrolled differences in the level of food restriction
or consumption cannot explain behavioral differences,
since increasing the level of food restriction by decreasing
body weight from 0 to 20% had no significant impact on
either the proportions of good and poor decision makers or
the evolution of their behavior (Rivalan et al. 2009a). The
use of a palatable food pellet (Formula P, Sandow) cer-
tainly contributes to this effect.
Importantly, we previously showed that the contrast
between one pellet (favorable options) vs two (unfavorable
options) is a crucial aspect of the test since differences
between PD and GD in the amount of work they are able to
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Brain Struct Funct (2018) 223:897–912
provide for food is observed for obtaining two pellets at a
time, but not for one (Rivalan et al. 2009a). Conversely, all
rats behave similarly in a version of the task where all
options provide the same number of rewards, but deliver
the four different penalties. That is, they prefer the shorter
penalty (6 s), followed by the 12 s long one and avoid the
long penalties.
The apparent large difference between short and long
penalties only results in a ratio 5 between the two kinds of
options. When the duration of the longer penalties is
decreased (Rivalan et al. 2009a) reducing the ratio to 4 or
3, poor decision-maker behavior is unchanged, suggesting
that the contrast between the rewards is fundamental and
drives their behavior. Interestingly, good decision-makers
take more time to select advantageous options because of
an increased difficulty of the task. As a whole, the com-
bination of the parameters used in the standard RGT allows
obtaining very similar results to those obtained in the
human IGT (Bechara et al. 2002).
In fact, poor decision-making in rats can be explained by
their particular combination of behavioral and cognitive
traits recently demonstrated: risk-taking, reward-seeking,
motor impulsivity and behavioral inflexibility (Rivalan
et al. 2013). This combination is reminiscent of several
mental disorders such as pathological gambling and
addiction (van den Bos et al. 2013). It appears to be a
consequence of an over-valuation of high-reward-high-risk
options in the task that cannot be modulated by learning
because of their inflexible behavior. Thus, it demonstrates a
strong alteration in long-term outcome evaluation and/or
insensitivity to future long-term consequences as proposed
by Damasio and Bechara in humans (Bechara et al. 1997;
Fitoussi et al. 2015; Rivalan et al. 2009a, 2011, 2013).
Our study shows that all poor decision-makers in the
RGT exhibit behavioral inflexibility, as well as a small
fraction of good decision-makers, confirming previous data
(Rivalan et al. 2013). Behavioral flexibility was tested in a
RGT-reversed version, during which advantageous and
disadvantageous options were spatially swapped. In this
RGT variant, shifting behavioral responses based on pre-
vious learned associations is necessary for adaptation to
changes, such as in standard simple reversal procedures
(Floresco et al. 2009). Cognitive processes engaged for
RGT solving can be highly predicted by a combination of
several core behavioral features, including not only
behavioral flexibility but also motor impulsivity, risk-tak-
ing and reward-seeking (Rivalan et al. 2013). This may
explain why poor performers in the RGT-reversed version
may not also be poor performers in the RGT: some
inflexible good decision-makers, being for example less
impulsive, or less sensitive to risk or reward. It may explain
why a relationship between IGT performances and reversal
Brain Struct Funct (2018) 223:897–912
learning in humans has only been sparsely reported (Clark
et al. 2004; Toplak et al. 2010).
Here, we show an additional cognitive process related to
inflexible behavior and its putative contribution for deci-
sion-making. Inflexible rats were characterized by an
altered efficiency in goal-directed behavior during a con-
tingency degradation procedure, independently of learning
abilities during training. The less flexible it was in shifting
choices in the RGT-reversed version, the less sensitive to
the contingency degradation the animal was. This rela-
tionship was observed for both inflexible good and poor
decision makers, suggesting that their inflexible behavior in
the reversal RGT procedure may have similar origins.
Thus, it seems reasonable to propose that during the
reversal, poor decision makers failed to adapt to contin-
gency reversal rather than suddenly preferring the advan-
tageous options. Inter-individual differences in the
sensitivity to contingency degradation could be revealed by
ensuring reduction of a non-specific parameter: variations
in general motor activity. For this purpose, a strict adjust-
ment of test conditions according to individual level of
motor activity during training was applied (see ‘‘Materials
and methods’’).
In the present study, we also assessed rats’s ability to
adapt their responding to a change in instrumental contin-
gency (Balleine and Dickinson 1998) using a random food
delivery procedure (Coutureau et al. 2012; Yin et al. 2006),
yoked to training food delivery to avoid a bias related to
inter-individual differences in activity levels. Under these
circumstances, some rats showed a marked decrement of
responding suggesting that they were sensitive to the
consequences of their action. Such a decrease in lever press
performance is generally assumed to result from a
decreased A–O contingency computation. It must be
acknowledged, however, that this suppression of lever
pressing may also occur due to acquisition of reward-re-
lated approach behavior to the magazine, as our results
suggest. By contrast, however, inflexible rats maintain a
high level of operant behavior throughout the contingency
changes, suggesting that they rely on habitual (S-R)
responding. Such an instrumental profile may be related to
the ‘‘illusion of control’’ that human pathological gamblers
show (Orgaz et al. 2013). It may also be related to perse-
verative and compulsive-like behaviors observed in
inflexible rats using different impulsivity schedules. Inap-
propriate compulsive behaviors (Dalley et al. 2011) may
result from attributing excessive incentive value to reward-
associated stimuli (Berridge and Robinson 1998; Flagel
et al. 2009).
909
Ability to update the A–O association and its
neurobiological correlates
In the second part of our experiment, we aimed to identify
the neural correlates of inter-individual differences in
sensitivity to contingency degradation using Zif268 as a
marker of neuronal reactivity. We report opposing corre-
lations between the ability to update the A–O association
during contingency degradation and Zif268 expression in
the mPFC and the dorsal striatum. A poor sensitivity to
contingency degradation was associated with a lower
expression of Zif268 in the mPFC (IL and PL) and a higher
one in the dorsal striatum (DLS and DMS). As illustrated
by the ratio of reactivity between the two types of struc-
tures, this interplay would play an essential role for
response adaptation in a changing environment. No clear
dissociation could be observed between activation of IL
and PL nor between activations of the lateral and the
medial parts of the dorsal striatum, related to the ability to
update A–O association, although their respective roles
have often been dissociated.
It is possible that differences in motor activity between
groups may have contributed to these differences in brain
activities. However, the absence of correlation between
Zif268 expression ratios between prefrontal and striatal
structures, and motor index, in contrast to sensitivity to
degradation, indicates that the immunoreactivity differences
cannot solely be attributed to unspecific motor output.
Within the dorsal striatum, the DLS and DMS differ in their
interconnectivity, receptor distribution and mechanisms of
plasticity (Yin and Knowlton 2006). The DMS receives multi-
modal information from the medial PFC and CgA, whereas
the DLS predominantly, but not exclusively, receives senso-
rimotor information from the sensorimotor cortex (Yin and
Knowlton 2006). The DMS (specifically the posterior region)
is more traditionally linked with goal-directed behavior effi-
ciency (Yin et al. 2005), specifically when considering the
causal relationship between an action and its outcome;
whereas the DLS (notably the posterior part) is often linked to
habits formation (Tricomi et al. 2009) and thus, specific
stimuli-response (S-R) association. However, some studies
found difficulties in dissociating the contribution of the DMS
vs DLS, e.g., goal-directed vs habitual-like behaviors (Stal-
naker et al. 2010) and consequently, their respective roles have
been reconsidered. A hypothesis is that a coexistence of S-R
and A–O information processing within these two striatal
subparts may occur (Stalnaker et al. 2010; Williams and
Eskandar 2006; Yin and Knowlton 2006). This hypothesis is
supported by our data in which DMS and DLS show similar
patterns of activation in our subgroups.
The general decrease in instrumental responses
throughout the contingency degradation test sessions may
reflect flexible, goal-directed behavior. However, this
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decrease is much slower in rats that are less sensitive to
contingency degradation and we cannot exclude that these
rats, that exhibited a high level of lever presses, were
partially driven by a habit mode, classically attributed to
DLS activity. Thus, a transition from a habit-becoming
behavior toward goal-directed behavior could occur during
the contingency degradation task, explaining a progressive
disengagement of DLS as operant behavior decreases in
rats with sensitivity to contingency degradation. Indeed,
the higher the sensitivity to contingency degradation, the
lower the Zif268 reactivity in DLSa. Differences were also
significant in posterior parts of DLS and DMS according to
this sensitivity, indicating a disengagement of the dorsal
striatum once the A–O association has been updated. The
similar level of control rat’s activity in the DLS to that of
sensitive individuals corroborates this hypothesis. These
results are in line with recent data showing the role of DMS
Zif268 activation in the transition from goal-directed
responding to habit-based actions (Maroteaux et al. 2014).
Indeed, Zif268 was shown to decrease in DMS once the
task performed was learned, but remained high in DLS.
Thus, the higher DMS activation observed in less sensitive
rats could reflect the active transition from habit-based
behavior to goal-directed responding, whereas the higher
DLS activation would reflect the largest habit component
of their behavior.
Interestingly, our data indicate that an efficient update of
A–O contingency is associated with a higher engagement
of the mPFC, notably in PL and IL. These two PFC areas,
strongly interconnected (Vertes 2004), are crucial for
coordinating actions and habits. Higher PL activation in
FLEX-S could, therefore, mediate rats’ ability to detect A–
O contingency variations (Corbit and Balleine 2003;
Coutureau et al. 2012; Naneix et al. 2009). Indeed, PL is a
key component of the neural circuit regulating goal-di-
rected behavior in rodents and primates, both for encoding
A–O contingency and for the representation of the outcome
as a goal (Balleine and O’Doherty 2010; Killcross and
Coutureau 2003). At the same time, the higher IL activa-
tion in rats sensitive to contingency degradation could
mediate the suppression of learned A–O association
required to extinguish behavior. This is in accordance with
recent views proposing that IL plays a key role in the
regulation of flexible behaviors and in inhibiting estab-
lished A–O association, when no more reward is delivered,
i.e., in extinction processes (Millan et al. 2011; Peters et al.
2008). Thus, the involvement of IL in the contingency
degradation paradigm may share some similarities with
that of standard extinction tests. It is noticeable that PL and
IL activities do not rely on activity levels, since active
control and FLEX-S rats with a low level of activity have
similar IL and PL activations.
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Brain Struct Funct (2018) 223:897–912
Finally, it is worth noting that human studies have
reported individual differences in cortico-striatal connec-
tivity related to the balance between goal-directed and
habitual actions (de Wit et al. 2012). Vulnerability to
habitual actions was related to premotor cortex—posterior
putamen connectivity, whereas flexible goal-directed
action was predicted by medio-ventral PFC—caudate
connectivity. It could, therefore, be hypothesized that our
results in rats may also originate from differences in
functional connectivities that remain to be explored.
Implication for decision-making-related disorders
Poor decision-makers are characterized by behavioral
inflexibility in the RGT reversal procedure, as well as
perseverative and compulsive-like behaviors (Rivalan et al.
2013), some behavioral traits that are reminiscent of
symptoms of several executive disorders. Indeed, perse-
verative responses are typically observed after acute
administration of psychostimulants (Evenden and Ko 2005)
and inflexible and compulsive behaviors are observed in
drug addiction (Calu et al. 2007; Jentsch et al. 2002),
pathological gambling (Goudriaan et al. 2006) and obses-
sive–compulsive disorder (OCD) (DSM-V 2013; Gillan
et al. 2011). Here, we show that inflexible behavior of poor
decision-makers (but also some good decision-makers)
could be related to a less-efficient encoding of goal-di-
rected action–outcome (A–O) associations in a changing
environment that could then contribute to the etiology of
these disorders. This deficit, related to mPFC/dorsal striatal
imbalance highlights key cognitive mechanisms that could
play a pivotal role in several psychiatric disorders. For
example, the critical transition from goal-directed to
habitual behaviors is a predominant hypothesis that may
explain, at least in part, addictive behavior (Everitt and
Robbins 2013) or OCD (Gillan et al. 2011). We hypothe-
sise that this lower cognitive efficiency—subserved by a
specific mPFC/dorsal striatal imbalanced activation, and/or
malfunction—may promote maladaptive habit formation
among poor decision-makers (Killcross and Coutureau
2003). As such, it could represent a key marker of vul-
nerability for decision-making-related disorders, such as
addiction.
Conclusion
In this study, we found that inflexible behavior as shown
predominantly in poor decision-makers is related to an
impaired ability to update an A–O association that medi-
ates goal-directed behavior. Taken together, our results
demonstrate that inter-individual differences in the transi-
tion between goal-directed and habit-based behaviors can
Brain Struct Funct (2018) 223:897–912
be revealed in a normal population of rats and offer
important and novel insights on how information is pro-
cessed through the cortico-striatal network. Although these
inter-individual differences in behavior are subtle, unlike
comparisons between standard experimental and control
groups, the differential approach tends to bring together
animal research and human psychological concepts (Ri-
valan et al. 2009b). By focusing on healthy individuals in a
non-invasive manner, this work contributes with others
(Bardo et al. 1996; Belin and Deroche-Gamonet 2012;
Blondeau and Dellu-Hagedorn 2007; Deroche-Gamonet
et al. 2004; Kabbaj and Akil 2001; Koolhaas et al. 2007;
LaPorte et al. 2010; Matzel and Kolata 2010; Rivalan et al.
2009b; Robbins et al. 2012; Robinson et al. 2014) to
demonstrate the relevance for identifying potential
biomarkers that could not only promote decision-making-
related disorders such as addiction, and OCD, but also
neural developmental disorders (i.e., developmental coor-
dination disorder) in which transitions between goal-di-
rected behavior and habits are disrupted. This work is in
line with the recent proposal by Robbins et al. (2012) to
undertake a more objective description of psychiatric dis-
orders through predisposing traits and neurocognitive
endophenotypes, thereby explaining the high level of
comorbidities between mental disorders.
Acknowledgements This work was supported and funded by the
Centre National de la Recherche Scientifique, the University of
Bordeaux and the Conseil Régional d’Aquitaine. We thank B. Lor-
geoux, S. Lelgouach, A. Fayoux for technical assistance, A. Marc-
hand, S. Parkes and F. Naneix for their helpful comments on the
manuscript and editorial assistance.
Compliance with ethical standards
Conflict of interest The authors declare no potential conflicts of
interest.
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