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Genes, Brain and Behavior (2010) 9: 103109

2009 The Authors


Journal compilation 2009 Blackwell Publishing Ltd/International Behavioural and Neural Genetics Society

The D2 dopamine receptor gene variant C957T affects


human fear conditioning and aversive priming
E. Huertas , G. Ponce, , M. A. Koeneke, ,
, , T. Palomo, ,

C. Poch , L. Espana-Serrano
,

M. A. Jimenez-Arriero
and J. Hoenicka,,
Facultad de Psicologa, Universidad Complutense de Madrid,
Campus de Somosaguas, Madrid 28223, Spain; Hospital
Universitario 12 de Octubre, Servicio de Psiquiatra, Avda. de

Cordoba
s/n, Madrid 28041, Spain; Centro de Investigacion

Biomedica
en Red de Salud Mental (CIBERSAM), Spain
*Corresponding author: J. Hoenicka, Servicio de Psiquiatra,

Hospital Universitario 12 de Octubre, AV de Cordoba


SN, Madrid
28041, Spain. E-mail: jhoenicka@gmail.com

Polymorphisms of DRD2 and ANKK1 have been associated with psychiatric syndromes where there is believed
to be an underlying learning process deficit such as
addiction, post-traumatic stress disorder and psychopathy. We investigated the effects of the DRD2 C957T and
ANKK1 Taq IA single nucleotide polymorphism (SNP),
which have been associated with psychopathic traits
in alcoholic patients, on fear conditioning and aversive
priming in healthy volunteers. We found that the DRD2
C957T SNP, but not the ANKK1 Taq IA SNP, was associated with both differential conditioning of the skin
conductance response and the aversive priming effect.
There were no differences between the genotype groups
with respect to the extinction of the skin-conductance
conditioned response. These results suggest that the
C957T SNP could be related to learning differences associated with the risk of developing psychiatric disorders
in individuals that are carriers of the C homozygous
genotype. Our genetic data raise the possibility that the
dopaminergic system functional variations determined
by this SNP could affect fear learning.
Keywords: ANKK1, aversive priming, cognitive bias, DRD2,
fear conditioning, polymorphism

Received 16 June 2009, revised 01 September 2009, 30


September 2009, accepted for publication 01 October 2009

The study of biological and environmental influences on psychopathic personality has indicated that individual differences
in antisocial behaviour are highly heritable (Fontaine et al .
2008; Viding et al . 2005). In clinical samples, the clearest link
between genetic variation and psychopathic traits has been
established with genes of the locus 11q22-q23 (Ponce et al .
2008). The most recent data show that the single nucleotide
polymorphisms (SNPs) of the D2 dopamine receptor gene
(DRD2 ) and the ankyrin repeat and kinase domain 1 (ANKK1),
doi: 10.1111/j.1601-183X.2009.00543.x

C957T (rs6277) and Taq IA (rs1800497), respectively, are


epistatically associated with psychopathic traits in alcoholic
patients (Ponce et al . 2008). The DRD2 gene (OMIM #
126450) encodes a G protein-coupled receptor which plays
a central role in reward and learning mechanisms (Hoenicka
et al . 2007), and its C957T SNP has also been associated with
schizophrenia (Hoenicka et al . 2006; Lawford et al . 2005) and
post-traumatic stress disorder (PTSD) (Voisey et al . 2009).
The ANKK1 gene (OMIM # 608774), which is adjacent to
DRD2, codes for a novel predicted kinase (Neville et al . 2004).
The ANKK1 Taq IA SNP, which is also a DRD2 gene marker,
has been widely associated with addiction, especially with
severe and antisocial alcoholism (Noble 2000; Ponce et al .
2003), thus suggesting a role for this SNP in reward and
learning systems. These DRD2 and ANKK1 gene variants
have also been studied in different learning paradigms. In
healthy volunteers, C957T has been linked to reinforcement
learning (Frank et al . 2007). Taq IA has been associated with
high reward sensitivity (Lee et al . 2007), the capacity of
learning from errors (Klein et al . 2007), a greater sensitivity
to negative feedback in neuropsychological tasks (Althaus
et al . 2009) and reversal learning (Jocham et al . 2009).
Psychopathy has been associated with significant impairment in several types of learning. Individuals with psychopathy show impaired stimulus-reinforcement learning in
the context of aversive conditioning (Birbaumer et al . 2005;
Flor et al . 2002). However, Ishikawa et al . (2001) reported
increased cardiovascular reactivity in stressed successful
psychopaths and they suggest that this higher response
could help to prevent risk behaviours. Thus, psychopathy could essentially entail a specific deficit in emotional
response to the harm suffered by others and not a general
hyporeactivity to threat.
These data, together with the molecular findings at locus
11q22-q23 in clinical and control populations, suggest that
some aversive learning paradigms could be endophenotypes
linked to DRD2 and ANKK1 gene variants. For example, fear
conditioning and aversive priming are potential endophenotypes that could be involved in the pathophysiology of
certain psychiatric disorders associated with learning abnormalities, such as psychopathy and PTSD. Fear conditioning is
a basic form of associative learning involved in the emotional
response to threats, and aversive priming is a bias favouring identification of threats after an aggression. Because
genetic, neuroimaging and clinical evidence indicates that
ANKK1 and DRD2 polymorphisms may play an important
role in affective information processing, reactivity to stress
and fear learning, we investigated the relationships between
both DRD2 C957T and ANKK1 Taq IA, and psychophysiological and behavioural indices of fear to examine whether these
SNPs are related to fear conditioning and aversive priming.

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Methods
Participants
Sixty-three healthy undergraduate students (32 females) at the Universidad Complutense de Madrid, Madrid, Spain, were recruited for
the study. Their age range was 1927 years and they received 20
as a monetary incentive. Data from one participant were excluded
because the genetic analysis could not be completed, and data from
a further two participants were excluded because of extreme values
on the recognition time task. The study was approved by the Ethics
Committee of Hospital 12 de Octubre, Madrid. Participants signed
two informed consent forms, one for the experimental procedure
and one for the genetic study.

Aversive learning experiment


The experimental study was carried out in the Faculty of Psychology
of the Universidad Complutense de Madrid. The participant was
seated in a sound-attenuated and tenuously illuminated experimental
room that was connected to an adjacent control room.

Stimulus materials
During the conditioning phase, four pictures of neutral black-andwhite faces (2 male, 2 female) from the Eckman and Friesen set
(numbers 21, 41, 65 and 99) were presented to each participant. Half
of the participants were presented the two male faces as conditioned
stimuli (CS+ and CS) and the two female faces served as additional
faces to familiarize the participant with them. For the remaining
participants, the faces were presented in the opposite manner.
The face that was coupled to the CS+ was also balanced among
participants. During the priming phase, these four faces were used
as those to be recognized (old faces). In addition, 10 neutral faces
from the Eckman and Friesen set (numbers 6, 13, 28, 33, 47, 56, 72,
83, 92 and 110) were used as distractors (distractor-faces). The faces
were projected from the control room on a screen located about
110 cm in front of the participant. The image size was 22 30 cm.
Electric shocks were generated by a Mark 100 stimulator (Farrall
Instruments, Inc., Grand Island, NE, USA) fed by batteries and
optically isolated from the computer. The electric shocks were applied
to the internal surface of the forearm through a Tursky concentric
electrode. The intensity of the shock was fixed by the participant
following the instruction it must be clearly uncomfortable but not
painful. During the conditioning phase, shocks were applied to the
dominant forearm for 200 milliseconds, and during the priming phase
they were applied to the non-dominant forearm for 180 milliseconds.
Neutral tones were 1000 Hz and the intensity was set by the
participant following the instruction you must hear it clearly, but it
must not be unpleasant at all. The tones were presented for 200
milliseconds during the conditioning phase and for 180 milliseconds
during the priming phase.

Psychophysiological and behavioural recordings


The skin conductance response (SCR) was recorded using two
AgAgCl Beckman electrodes (8-mm diameter), filled with a 0.05 M
NaCl gel. Electrodes were fastened by adhesive collars to the palmar
surface of the second phalanges of the first and second fingers of the
non-dominant hand. The signal was amplified by an S7122 bioamplifier (Coulbourn Instruments, Whitehall, PA, USA) and sampled at
50 Hz. The SCR was scored as the maximum increase in conductance between the first and the fourth second after the onset of the
stimulus. The SCR value was considered zero if changes were lower
than 0.01 S or no increase was observed. The reaction time (RT)
was registered by a locally built console with two buttons, marked
yes and no.
An IBM-compatible personal computer was used to monitor the
presentation of stimuli, and to acquire and store the SCR and RT data.
It was equipped with a PCL-812 PG Multi-Lab card and a PCLD-785
Relay Output Board card (both from Advantech Co., Cincinnati, OH,
USA). The software was developed by the Technical Service of the
Faculty of Psychology of the Universidad Complutense de Madrid.

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Procedure
The experiment had four phases: presentation of distractors,
habituation, conditioning and aversive priming.
Upon arrival, the participants were reminded that they could leave
the experiment at any time. They were fitted with skin conductance
electrodes, and then they viewed each of the 10 faces that would
operate as distractors in the priming phase two times. Each picture
was presented for 8 seconds and the inter-trial interval (ITI) was
4 seconds. Next, the shock electrodes were fitted, and the intensity
of the shock and the tone were set. During the breaks in the
experiment, participants were asked again about the unpleasantness
of the stimuli and the intensity was modified when necessary. The
participant could also ask to change the intensity at any time.
Next, the participants were informed that they would view four
new faces several times each, and that one of the faces would
sometimes be paired with the shock (CS+) and the other would
sometimes be paired with the tone (CS). During the habituation
phase, the procedure included a non-reinforced presentation of
each of the CS+, CS and the two other old faces. During the
acquisition phase, the procedure included eight presentations of
the CS+ paired with the aversive shock and eight presentations
of the CS paired with the neutral tone (paired trials), three
presentations of each of the CS+ and the CS alone (test
trials), and four presentations of each of the other two old
faces.
The set habituation-conditioning was organized in six blocks with
a total of 34 trials and a single break after the 20th trial. The first
block was the habituation phase. The second block consisted of two
paired trials of the CS+ and two paired trials of the CS. Blocks 3, 4
and 5 consisted of two paired trials of each of the CS+ and the CS,
one test trial of each of the CS+ and the CS, and one presentation
of each of the other two faces. The sixth block consisted of one
presentation of each of these last two faces. Within the blocks,
the trials of each type were presented in semi-random order, with
the restriction that none of the faces appeared in more than three
consecutive trials. The presentation time for the faces was 8 seconds,
the interval between the appearance of the face and the onset of
shock or tone [stimulus onset asynchrony (SOA)] was 3 seconds and
the duration of the shock and the tone was 200 milliseconds. The ITI
varied randomly between 16 and 20 seconds.
Next, the aversive priming (Huertas-Rodrguez 1991) phase began.
Participants were informed that they would see several faces and
should indicate as quickly as possible, but trying not to make
mistakes, if the face was one of the four faces presented during
the conditioning phase or one of the faces seen at the beginning
of the experiment. It was therefore a recognition task, and not
only a familiarity-judgement task. Participants were warned that any
face could be preceded by the neutral tone, the aversive shock or
neither of them. The recognition decision was made by pressing the
corresponding buttons on the console. By pressing the button, the
face disappeared. When the participant did not press any button in a
period of 3 seconds after the face onset, it disappeared automatically.
The SOA was 190 milliseconds. The duration of the prime (shock or
tone) was 180 milliseconds. The ITI varied randomly between 19 and
21 seconds.
Participants underwent two presentations of the CS+ and two of
the CS, seven presentations of the other two old faces and 10
presentations of the distractors. The CSs were preceded once by the
aversive prime, and once by the neutral one. The rest of the faces
were preceded four times by the aversive prime, seven times by the
neutral prime and six times by neither of them. There was one presentation of each primeCS sequence, because the aversive-prime/CS
sequence and neutral-prime/CS+ sequence could operate as counterconditioning trials, thus reducing or eliminating the expected effect in
subsequent presentations. After trial 14, a break of 4 min was taken.
The four critical prime/CS sequences were presented in trials 9, 13,
17 and 20. There were four presentation orders for these sequences,
according to the eight possible permutations that fulfill the requirement for an alternate conditioned stimulus (CS+ and CS) and prime
(aversive and neutral) presentations. The presentation order for each
participant was random.
The priming phase finished with a presentation of each CS+
and CS without being preceded by shock or tone. The order of
presentation was counterbalanced between the participants.
Genes, Brain and Behavior (2010) 9: 103109

DRD2 C957T SNP affects fear learning

Genotyping
Samples from epithelial cheek cells were collected with buccal
swabs. Genomic DNA extraction and amplification were performed
using the illustra GenomiPhi V2 DNA Amplification Kit (GE Healthcare Europe, Barcelona, Spain) following the manufacturers protocol.
Genotyping was performed by Taqman assays, which were designed
to run on an ABI 7900HT machine with Sequence Detection System
software (Applied Biosystems, Foster City, CA, USA). In a previous
work, we evaluated the non-random association of the two SNPs
and found a low degree of linkage disequilibrium between Taq IA
and C957T in the control population (D = 0.58, r 2 = 0.14) (Ponce
et al . 2008).
The sequence-specific primers for ANKK1 Taq IA (5 CTGCCTCGAC
CAGC 3 and 5 CTGCCTTGACCAGC 3 ) were designed for the
C (A2) and T (A1) alleles, respectively. A common reverse primer
(5 -GCAACACAGCCATCCTCAAAG-3 ) was used. Five samples for
each genotype were confirmed by direct sequencing analysis. The
resulting genotypes for ANKK1 Taq IA were clustered according to
the presence of at least one A1 allele (A1+ genotype, A1 allele
homozygous and heterozygous; A1 genotype, homozygous for
the A2 allele) as previously described (Blum et al . 1990). DRD2
C957T was analysed as previously described by Hoenicka et al .
(2006). Briefly, the sequence-specific primers for the Taqman assays
(5 -CTGTCGGGAGTGCTG-3 and 5 -CTGTCAGGAGTGCTG-3) were
used for the C and T alleles, respectively, as was the common
reverse primer 5 -GCCCATTCTTCTCTGGTTTGG-3 . The genotypes
obtained were grouped assuming a recessive model for the C957
allele: homozygous individuals for the C allele vs. heterozygous and
homozygous individuals for the T allele.
We performed the HardyWeinberg equilibrium test using the
Haploview software (version 3.2; Whitehead Institute for Biomedical
Research; http://www.broad.mit.edu/mpg/haploview/index.php) and
we found no deviation from either in ANKK1 Taq IA (P = 0.44) or in
DRD2 C957T (P = 0.83). In addition, we observed in this sample the
same ANKK1 and DRD2 genotype distributions that were previously
found for both Spanish (Ponce et al . 2008) and European (NCBI:
http://www.ncbi.nlm.nih.gov/) healthy populations.

Statistical analysis
All SCR magnitudes were range corrected, by dividing each value
by the mean of the participants three maximal responses and
multiplying the result by 100 (PC SCR). A square-root transformation
was also applied to help to normalize the distribution [SQRT (1 + PC
SCR)]. In the case of the RTs, data were log transformed. However,
means are presented as milliseconds to allow a better interpretation.
The incorrect recognition responses (4.6% of total data) and RT
values over the grand mean 2.5 SD (1.7% of total data) were
replaced by the mean of the participant.
A repeated-measures general linear model (SPSS 15.0 for
Windows, SPSS Inc., Chicago, IL, USA) was used for each SNP,
with SCR and RT as consecutive dependent variables. We used a
two-tailed t -test for post hoc comparisons between genotypes, and
a one-tailed t -test for the comparisons for which we had a specific
prediction a priori. Given the unequal size of the genetically defined
groups (C957T SNP: CC group = 9, CT/TT group = 51; Taq IA SNP:
A1 = 18, A1+ = 42), additional MannWhitney tests were used
for post hoc comparisons between genotypes. This analysis gave
rise to an identical pattern of results to that of the t -test.

C957T genotype. Only the CS test-trial genotype linear


interaction achieved significance (F1.58 = 5.687; P = 0.020)
(Fig. 1b). Next, we analysed the test-trial genotype linear
interaction for the CS+ and CS separately. Only the interaction for the CS+ was significant (F1.58 = 8.401; P = 0.005;
partial 2 = 0.127). The CT/TT group exhibited a lower SCR
in the third trial when compared with the first trial (t = 2.975;
df = 50; P = 0.003), whereas the SCR in the CC group not
only failed to show a decrement but rather tended to increase
(P = 0.073). Thus, differential conditioning of SCR was higher
in the CC carriers during the acquisition phase. In these individuals, the response to CS+ did not decrease during the
conditioning process. Moreover, this divergence between
the genotype groups was not because of a difference in the
differential SCR during the habituation phase (P = 0.490),
or a difference in the SCR to the unconditioned stimulus
(P = 0.794), or a difference in electrodermal responsivity
(P = 0.453).
With respect to the Taq IA polymorphism, the repeated
measures ANOVA did not show a CS test-trial genotype
interaction (P = 0.688) (Fig. 1c).

Extinction
The CS genotype interaction analysis showed no significant
differences between groups in the extinction of the SCR
for the C957T SNP (P = 0.782) (Fig. 1b) or for the Taq I-A
(P = 0.147) (Fig. 1c).

Relationship between the C957T and TaqI-A SNPs


and aversive priming
We first confirmed an overall aversive priming effect. When
all the data were taken into account, there was a significant
prime CS interaction (F1.59 = 7.867; P = 0.007). The RT in
the aversive-prime/CS+ condition was shorter when compared with each of the other three conditions (P < 0.001).
When the C957T genotype was included in the model,
a significant prime CS C957T interaction pattern was
identified (F1.58 = 4.337; P = 0.042) (Fig. 2b). In the aversiveprime/CS+ condition, but not in the other three conditions,
the CC carriers showed a shorter RT than the CT/TT group
(t = 2.392, df = 58, P = 0.02; Cohen s d = 0.879). Thus,
after the differential conditioning process, the presence of an
aversive primer facilitates CS+ recognition, and this effect is
more prominent in the CC carriers.
When these analyses were performed by grouping according to the Taq IA SNP genotype, we did not find a significant
prime CS Taq IA interaction (P = 0.538) (Fig. 2c).

Results
Discussion
Relationship between the C957T and TaqIA SNPs,
and fear conditioning
Acquisition
The SCR analysis showed a significant CS test-trial C957T
interaction (F2.116 = 3.154; P = 0.046). This interaction effect
was subsequently parsed into orthogonal linear and quadratic
contrasts to evaluate the effect of the test-trial on the differential response to the CS+ and the CS depending on the
Genes, Brain and Behavior (2010) 9: 103109

We found that DRD2 C957T, but not ANKK1 Taq IA, was
associated with both SCR differential conditioning and
aversive priming in healthy participants. Specifically, during
acquisition, the CT/TT group exhibited a continued decrease
in the SCR to the CS+ with repeated presentation of the
stimulus, whereas the response of the C homozygous
carriers was maintained or tended to increase. On the

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Huertas et al.

Figure 1: Conditioning of the skin conductance response as a function of genotype. (a) The differential conditioning paradigm;
(bc) Relationship between the genotypes (DRD2 C957T SNP ; ANKK1 Taq IA SNP) and the magnitude of the SCR to the CS+ and
CS in the three acquisition test trials and the extinction trial. Asterisks indicate significant differences: *P < 0.05; **P < 0.01. SQRT
[1 + PC SCR]: squared root of 1 plus the percentage of the skin conductance response respect to the mean of the participants three
maximal responses; 1,2,3: test trials of acquisition; e: extinction.

contrary, when we examined the extinction of the CS+


response, we found no differences between the genotype
groups. Thus, the higher physiological response did not seem
to be associated with a learning impairment; instead, it seems
that the C homozygous carriers did not adapt to the CS+
exposure, as long as this stimulus represented a threat, thus
producing a sustained emotional response.
With respect to aversive priming, that has a clear similarity to cognitive biases associated with anxiety, our paradigm
assesses the speed of selective attention towards the threatening stimulus. After the differential conditioning process,
the C homozygous group had a shorter recognition time
in the aversive-prime/CS+ condition, thus reflecting that
either the CS+ had become more threatening, or that priming
is facilitated.
Fear conditioning and aversive priming usually have an
adaptive function. Normal fear to a threat warns the person of danger and the aversive priming facilitates a fast
and accurate perception of real threats. However, excessively intense fears could have maladaptive consequences
and enhanced cognitive-affective biases can increase the
likelihood of processing an unreal threat that in turn would
increase anxiety. Therefore, our study may also have clinical

106

implications. Indeed, our results are in line with data and theories pertaining to psychiatric disorders associated to the C957
homozygous genotype such as the PTSD (Voisey et al . 2009),
schizophrenia (Hoenicka et al . 2006; Lawford et al . 2005)
and substance abuse (Perkins et al . 2008). For instance,
individuals with PTSD seem to be more conditionable than
trauma-exposed individuals without PTSD (Orr et al . 2000).
Further, several studies suggest that there is an increased
likelihood of flashbacks and intrusive phenomena related
to the traumatic experience in these patients as a consequence of a subsequent increase in arousal. This retroactive
effect occurs even if the arousal is artificially triggered [e.g.
(Bremner et al . 1997; Nixon and Bryant 2005)]. A similar phenomenon has been found in schizophrenia where stressful
situations raise the likelihood of hallucinations (Freeman and
Garety 2003). In the case of drug abuse, situations leading
to the increment of the arousal also increase the likelihood
of images and thoughts related to the substance of abuse
(Sinha 2008). Indeed, in the case of nicotine addiction, the
smokers homozygous for the C957 allele took more cigarette
puffs during negative vs. positive mood (Perkins et al . 2008).
On the contrary, our results in the C957 homozygous, a risk
genotype for psychopathic traits (Ponce et al . 2008), seem
not to be consistent with one of the most replicated findings
Genes, Brain and Behavior (2010) 9: 103109

DRD2 C957T SNP affects fear learning

Figure 2: Recognition time in the priming task as a function of genotype. (a) The priming procedure in the four conditions:
aversive-prime/CS+ (Av/CS+), aversive-prime/CS (Av/CS), neutral-prime/CS + (Ne/CS+) and neutral-prime/CS (Ne/CS); (bc)
Relationship between the genotypes (DRD2 C957T SNP; ANKK1 TaqIA SNP) and the recognition time of the CSs in the Av/CS+,
Av/CS, Ne/CS+ and Ne/CS sequences. Asterisks indicate significant differences: *P < 0.05.

in criminal psychopaths, i.e. electrodermal hyporeactivity in


the anticipation of aversive stimuli (Fowles 2000). However,
we must remember that our study was performed in healthy
volunteers. Heightened autonomic stress reactivity has also
been observed in successful psychopaths when compared
wiht both unsuccessful psychopaths and controls (Ishikawa
et al . 2001). On the other hand, in our previous work on alcoholic patients, the C homozygous genotype was associated
with psychopathy exclusively when the ANKK1 Taq IA A1
allele was present. Thus, the effect of DRD2 C957T effect
on learning processes may vary according to the biological
background. Poor modulation of the electrodermal response
has been related to a deficit in inhibitory control, a clinical trait
that may be linked to a variety of psychiatric disorders marked
by impulsivity or disinhibition (Taylor et al . 1999) . Thus, the
C homozygous genotype, which was linked to an autonomic
over-reaction in this study, could be related to lesser control
and greater reactive aggressiveness, which are characteristic
of addictive and antisocial personality disorders.
An increased conditioned behavioural response to threats
during conditioning acquisition has also been observed in
mice when amphetamine was withdrawn (Pezze et al . 2002).
This heightened response may correlate with differences
in the functioning of the nucleus accumbens (NAc),
which is involved not only in reward learning but also in
Genes, Brain and Behavior (2010) 9: 103109

aversive conditioning (Pezze et al . 2001). Although aversive


conditioning has mostly been related to functioning of the
amygdala and prefrontal cortex, it has been showed that the
NAc also has a relevant role. Elevated dopamine levels have
been observed in the NAc in response to aversive outcomes
(Kalivas and Duffy 1995; Robinson et al . 1987; Young 2004),
CSs that are predictive of such outcomes or exposure to the
conditioning context (Pezze et al . 2002; Young 2004; Young
et al . 1993, 1998). Dopamine signalling in the amygdala and
prefrontal cortex is mediated mainly by the D1 dopamine
receptor, whereas D2 is highly expressed in the striatum.
The C957T SNP has been associated with mRNA stability
and protein synthesis in vitro (Duan et al . 2003), as well as
with an increased D2 receptor density in several brain areas
(Hirvonen et al . 2009). Furthermore, in mice, overexpression
of the D2 receptor in the striatum has been found to affect
dopamine functioning in the prefrontal cortex, thus leading to
working memory deficits (Kellendonk et al . 2006). Therefore,
learning differences related to cortical function may be a
reflection of a primary striatal variation.
The limitations of this study are related to the nature
of association studies; therefore, our results must be
interpreted with caution. The strength of our results is
tempered by a relatively small sample size, especially in some
genotype subgroups. Thus, to validate our results, it would

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Huertas et al.

be desirable to replicate our genetic studies in independent


samples. On the other hand, our approach to measure the evidence of extinction is somewhat atypical, because it involved
the last two trials of the priming phase and the need for the
participant to press the button on the recognition test. Extinction tests generally involve presentation of the CS to the participant with no additional tasks that would increase the SCR
to both the CS+ and the CS. Despite these limitations, we
believe that our study has identified a genetic variant of the
dopaminergic system that is related to aversive learning, and
that this variant deserves greater attention. Future research
should clarify to what extent this increased differential SCR
was because of a lower inhibitory control of emotional
response to danger rather than to fear-enhanced learning.
We cannot disregard the potential role of ANKK1 Taq IA
in other learning processes related to psychopathic traits.
This polymorphism has been associated with reward-related
personality traits (Lee et al . 2007) and different learning
paradigms (Althaus et al . 2009; Klein et al . 2007). In any case,
the study of other aversive learning paradigms would shed
light on the role, if any, of the epistatic association between
C957T and Taq IA in learning impairment in psychopathy.
In summary, our results suggest that functional variations
in D2 signalling determined by the C957T SNP could be
related to learning differences associated with the risk of
developing psychiatric disorders in carriers of the C homozygous genotype.

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Acknowledgments
This work has been supported by the Fondo para Investigaciones
Sanitarias (FIS), Madrid, Spain, grants no. 05/0731 and 08/0529.
CIBERSAM is an initiative of the Instituto de Salud Carlos III.

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