Neuroscience Letters 503 (2011) 220–223

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Neuroscience Letters
journal homepage: www.elsevier.com/locate/neulet

Functional polymorphism of the GTP cyclohydrolase 1 gene affects the

personality trait of novelty seeking in healthy subjects
Ryoichi Sadahiro a , Akihito Suzuki a,∗ , Yoshihiko Matsumoto a , Naoshi Shibuya a ,
Masanori Enokido a , Mitsuhiro Kamata a , Kaoru Goto b , Koichi Otani a
a
Department of Psychiatry, Yamagata University School of Medicine, Yamagata, Japan
b
Department of Anatomy and Cell Biology, Yamagata University School of Medicine, Yamagata, Japan

a r t i c l e i n f o a b s t r a c t

Article history: GTP cyclohydrolase 1 (GCH1) is the initial and rate-limiting enzyme in the biosynthesis of tetrahydro-
Received 11 July 2011 biopterin, which is an essential cofactor for biosynthetic enzymes of dopamine, serotonin, and nitric oxide.
Received in revised form 10 August 2011 In the present study, the association of functional polymorphism of the GCH1 gene (C+243T, rs841) with
Accepted 20 August 2011
personality traits was examined in 902 healthy Japanese subjects. Personality traits were assessed by
the Temperament and Character Inventory (TCI), and the GCH1 genotype was detected by a PCR–RFLP
Keywords:
method. There were no significant main effects of the GCH1 genotype on the seven TCI dimension scores,
GCH1
but significant interaction effects between the GCH1 genotype and gender were found on the scores of
Polymorphism
Personality
novelty seeking. Post-hoc analysis revealed that males with the C/C genotype had higher scores of novelty
Novelty seeking seeking than those with the C/T genotype or those with the T/T genotype, while in females the scores of
novelty seeking were not different among the genotype groups. The present study thus suggests that the
C+243T polymorphism of the GCH1 gene affects the personality trait of novelty seeking in males.
© 2011 Elsevier Ireland Ltd. All rights reserved.

GTP cyclohydrolase 1 (GCH1) catalyzes the conversion of guano-
sine triphosphate to dihydroneopterin triphosphate, and is the
initial and rate-limiting enzyme in the biosynthesis of tetrahy-
drobiopterin (BH4) [8] (Fig. 1). BH4 is an essential cofactor for
biosynthetic enzymes of several neurotransmitters, i.e., tyrosine
hydroxylase, tryptophan hydroxylase, phenylalanine hydroxylase,
and nitric oxide (NO) synthase, which catalyze the production of
dopamine, serotonin, tyrosine, and NO, respectively [8] (Fig. 1).
It has been reported that inherited GCH1 deficient mice show
decreased levels of BH4, dopamine, serotonin, and norepinephrine
in brain [11] and decreased levels of NO in plasma [13]. Patients
with dopa-responsive dystonia or Segawa disease, which is char-
acterized by GCH1 deficiency caused by rare GCH1 gene mutations,
have been reported to show decreased levels of BH4 accom-
panied by low levels of the dopamine metabolite homovanillic
acid and the serotonin metabolite 5-hydroxyindoleacetic acid in
cerebrospinal fluid [11]. These patients also exhibit various psy-
chological symptoms such as mental retardation, depression, and
obsessive-compulsive disorders [8,10]. These studies suggest that
GCH1 activity is involved in human mental functions and behaviors.
∗ Corresponding author at: Department of Psychiatry, Yamagata University School
of Medicine, 2-2-2 Iidanishi, Yamagata 990-9585, Japan. Tel.: +81 23 628 5322;
fax: +81 23 628 5325.
E-mail address: suzukiakihito@hotmail.com (A. Suzuki).
Human GCH1 gene is located at 14q22.1–22.2 and contains 6
exons. Zhang et al. [22] reported a C+243T single nucleotide poly-
morphism (C59038T, rs841) in 3 untranslated region of the GCH1
gene. They showed that the T allele of this polymorphism caused
lower luciferase activity in transfected chromaffin cells than the C
allele [22]. Furthermore, the T allele was related to lower urinary
levels of NO, higher blood pressure and heart rate, and dysfunction
of the baroreceptor reflex in vivo [22]. This allele was also associ-
ated with lower plasma NO levels and increased risk of vascular
diseases in patients with type 2 diabetes [14]. Meanwhile, a hap-
lotype comprising 3 single nucleotide polymorphisms of the GCH1
gene, which is in strong linkage disequilibrium with the T allele
of the C+243T polymorphism [8], was associated with lower GCH1
mRNA levels [2,20], reduced BH4 levels in vascular and leucocytes
[2,20], and reduced pain sensitivity [20].
The Temperament and Character Inventory (TCI) is a compre-
hensive personality scale developed from biological perspectives
by Cloninger et al. [4]. The TCI has four temperament dimen-
sions (novelty seeking, harm avoidance, reward dependence, and
persistence), and three character dimensions (self-directedness,
cooperativeness, and self-transcendence). They hypothesized that
novelty seeking, harm avoidance, and reward dependence were
related to dopamine, serotonin, and norepinephrine, respectively
[4]. It has been suggested that several psychiatric disorders, such
as anxiety disorders, mood disorders, eating disorders, and sub-
stance dependence, are related to specific dimensions of the TCI

0304-3940/$ – see front matter © 2011 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.neulet.2011.08.040
 R. Sadahiro et al. / Neuroscience Letters 503 (2011) 220–223
Guanosine triphosphate
GCH1
Dihydroneopterin triphosphate
PTPS
6-pyruvoyl-tetrahydropterin Tyrosine
SR
Tetrahydrobiopterin TH
L-dopa
Dopamine
Fig. 1. Involvement of GCH1 in biosynthesis of dopamine, serotonin, and nitric oxide. GCH1, GTP cyclohydrolase 1; PTPS, 6-pyruvoyl-tetrahydropterin synthase; SR, sepiapterin
reductase; TH, tyrosine hydroxylase; TPH, tryptophan hydroxylase; PAH, phenylalanine hydroxylase; NOS, nitric oxide synthase.
[3,4,5,9]. Furthermore, prospective studies showed that high harm
avoidance predicted future depression in a general population [5,9].
Therefore, it is important to identify the factors involved in the char-
acterization of personality traits to promote understanding of the
pathogenesis of these disorders.
Taken these discussions all together, it is possible that the
C+243T polymorphism in the GCH1 gene affects personality traits.
However, there have been no studies examining the relationship
between this polymorphism and personality traits. Therefore, we
studied the association of this polymorphism with personality
traits in healthy subjects.
Originally, 995 unrelated Japanese volunteers without serious
physical diseases were recruited from medical students and the
staff at five hospitals. Forty-nine subjects with a current or past
history of psychiatric disorders according to the DSM-IV [1] were
excluded after interview by well-trained psychiatrists. In 14 of the
995 subjects, the GCH1 genotype was not determined due to fail-
ure of PCR amplification. Thirty subjects were excluded due to a
missing data. Thus, the data for 902 subjects were used for statisti-
cal analyses. Four hundred ninety seven subjects were males, and
405 were females. The mean ± SD of age was 27.1 ± 8.2 years. The
study protocol was approved by the Ethics Committee of Yamagata
University School of Medicine, and all subjects provided written
informed consent to participate.
Assessment of personality traits was performed using the
Japanese version of the TCI, which has been verified to have high
internal consistency and construct validity [12].
The 361 bp in the GCH1 gene was amplified using a set of primers
(forward; 5 - GTT GGT TGC CGA TCG TAC TG -3 and reverse; 5 -
CAG TAT ACT GGG CAC AGT TC -3 ) in 20 ␮l volume containing
100 ng of genomic DNA, 0.5 ␮M of each primer, 200 ␮M of each
dNTP, 1.5 mM of MgCl2 , and 1.25 U of HotStar Taq DNA polymerase
(Qiagen, Tokyo, Japan). After an initial denaturation step at 95 ◦ C
for 15 min, 40 cycles were performed at 94 ◦ C for 0.5 min, 54 ◦ C for
0.5 min, and 72 ◦ C for 0.5 min. Finally, an elongation step was per-
formed at 72 ◦ C for 10 min. Five ␮l of the PCR product was digested
overnight with 10 U of TaiI (Cosmo Bio Co, Tokyo, Japan). The frag-
ments were electrophoresed on a 2% agarose gel with ethidium
bromide staining and visualized using ultraviolet light. After diges-
tion by TaiI, the PCR product containing the C allele is cleaved
into 225- and 136 bp fragments, while that containing the T allele
remains intact (361 bp fragment).
The Hardy–Weinberg equilibrium for genotype distribution was
tested by the chi-square test. Zhang et al. [22] reported a significant
221
Tryptophan Phenylalanine Arginine
TPH PAH NOS
5-hydroxytryptophan Tyrosine Nitric oxide + citrulline
Serotonin
interaction effect between the GCH1 polymorphism and gender on
blood pressure. Thus, the effects of the GCH1 polymorphism on
the TCI dimension scores were tested by the two-factor analysis
of covariance with the GCH1 genotype and gender as factors and
with age as a covariate. In these analyses of seven TCI dimensions,
a p value less than 0.05/7 (two-tailed) was regarded as significant
according to Bonferroni correction. When the interaction between
the GCH1 polymorphism and gender was significant in some TCI
dimension, that dimension score was compared among the GCH1
genotype groups in males and females separately, by the one-factor
analysis of covariance with age as a covariate followed by the LSD
test. A p value less than 0.05 (two-tailed) was regarded as signif-
icant. All statistical analyses were performed using SPSS 14.0J for
Windows (SPSS Japan Inc, Tokyo, Japan).
The distribution of the GCH1 genotypes was in the
Hardy–Weinberg equilibrium (2 = 0.185, df = 1, p = 0.667)
(Table 1). There were no significant main effects of the GCH1
genotype on the seven TCI dimension scores, while significant
main effects of the gender were found on the scores of harm
avoidance (p < 0.007) and reward dependence (p < 0.007) (Table 1).
The interaction between the GCH1 genotype and gender was
significant (p = 0.014) on the scores of novelty seeking (Table 1).
In the post-hoc analysis, there were significant differences in the
scores of novelty seeking among the GCH1 genotype groups in
males (F = 6.432, p = 0.002) but not in females (F = 1.190, p = 0.305).
Males with the C/C genotype had higher scores of novelty seeking
than those with the C/T genotype (p = 0.002) or those with the T/T
genotype (p = 0.004) (Fig. 2).
In the present study, the T allele of the C+243T polymorphism
in the GCH1 gene was related to lower scores of novelty seek-
ing in males. It is described that high scorers of novelty seeking
are exploratory, curious, impulsive, extravagant, enthusiastic, and
disorderly, while low scorers are indifferent, reflective, frugal,
detached, orderly, and regimented [4]. The T allele of the GCH1
polymorphism was related to lower luciferase activity in trans-
fected chromaffin cells [22], while a GCH1 haplotype in strong
linkage disequilibrium with this allele [8] was associated with
lower GCH1 mRNA levels and reduced BH4 levels in vascular and
leucocytes [2,20]. These findings lead to the assumption that the
subjects with the T allele of the GCH1 polymorphism have lower
levels of dopamine, serotonin, and NO in brain. Thus, the present
results suggest that males with the GCH1 genotype predictive of
lower levels of these neurotransmitters have lower scores of nov-
elty seeking.
222 R. Sadahiro et al. / Neuroscience Letters 503 (2011) 220–223

Table 1
Effects of the GCH1 polymorphism and gender on TCI dimension scores.

No. Novelty Harm Reward Persistence Self-directedness Cooperativeness Self-transcendence

seeking avoidance dependence

Males, mean ± SD
C/C 185 22.9 ± 4.8 18.1 ± 6.2 15.5 ± 3.7 4.3 ± 1.7 28.9 ± 6.1 28.1 ± 4.9 8.7 ± 4.8
C/T 237 21.4 ± 5.1 18.1 ± 6.1 15.8 ± 3.6 4.4 ± 1.8 29.6 ± 6.2 27.8 ± 5.6 8.7 ± 4.6
T/T 75 21.0 ± 4.6 18.2 ± 5.5 15.7 ± 3.9 4.7 ± 1.8 29.4 ± 6.0 27.5 ± 5.3 8.9 ± 4.5
Females, mean ± SD
C/C 139 20.8 ± 5.1 20.0 ± 5.8 16.9 ± 3.3 3.9 ± 1.5 27.5 ± 7.0 28.9 ± 4.6 9.7 ± 4.1
C/T 191 21.4 ± 5.5 19.8 ± 5.9 16.8 ± 3.3 4.3 ± 1.8 28.3 ± 6.6 28.7 ± 4.6 9.4 ± 4.8
T/T 75 22.1 ± 5.8 19.3 ± 6.2 16.5 ± 3.4 4.3 ± 1.7 29.0 ± 6.5 28.7 ± 4.7 8.8 ± 4.5
Main effects, F
Genotype 0.801 0.133 0.223 4.020 1.765 0.336 0.370
Gender 0.430 12.817b 18.857b 4.501 4.810 6.971 2.739
Interaction effects, F
Genotype × Gender 6.155a 0.240 0.461 0.770 0.252 0.079 0.877

GCH1, GTP cyclohydrolase 1; TCI, Temperament and Character Inventory. P values were adjusted by Bonferroni correction for multiple testing.
a
p = 0.014.
b
p < 0.007.

It has been suggested that dopaminergic neurotransmission
is involved in novelty seeking [4]. Specifically, the relationship
between lower novelty seeking and higher synaptic dopamine lev-
els was supported by the genetic association studies on the tyrosine
hydroxylase [16] and monoamine oxidase A [19] polymorphisms.
Meanwhile, Reif et al. [15] showed that the males with the NO
synthase genotype predictive of higher NO levels exhibited lower
scores of excitement seeking of the NEO personality questionnaire
[7], which correspond to lower scores of novelty seeking of the
TCI [4]. These studies showing that either low dopamine or NO
levels induce higher scores of novelty seeking are opposite to the
present results. Since there have been no studies examining the
effects of the GCH1 polymorphism on these neurotransmitter lev-
els in brain, we cannot specify the exact mechanism. However, one
plausible possibility is that lower levels of dopamine, NO, and sero-
tonin caused by the GCH1 polymorphism may induce low scores of
novelty seeking. In fact, it has been reported that NO exerts a neg-
ative control over the levels of dopamine and serotonin [21], while
serotonin counteracts dopaminergic transmission by negative
feedback [6]. Thus, it is possible that these interactions among
dopamine, NO, and serotonin may be implicated in the results
observed in the present study.
The significant association between the GCH1 polymorphism
and novelty seeking was observed only in males, and none of the TCI
dimensions was affected by the GCH1 polymorphism in females.
It was reported that expression levels of GCH1 mRNA in brain
monoaminergic neurons were higher in male mice than in female
mice [18], probably because of modulating effects of sex hormones
on GCH1 expression [17]. Therefore, the sex-specificity observed in
the present study may be due to the effects of sex hormones on the
regulation of GCH1 activity.
In conclusion, the present study suggests that the C+243T poly-
morphism in the GCH1 gene affects the personality trait of novelty
seeking in males.
Conflict of interest

The authors declare no conflict of interest.

p=0.004
30
p=0.002 n.s. Acknowledgements

This study was supported by a Grant-in-Aid from the Global Cen-

ter of Excellence Program of the Japan Society for the Promotion of
Science, Japan, and by a funding from the Ministry of Education,
25
Culture, Sports, Science and Technology of Japan.
Novelty seeking (scores)

References

[1] American Psychiatric Association, Diagnostic and Statistical Manual of Mental

20 Disorders, 4th ed., American Psychiatric Association, Washington, DC, 1994.
[2] C. Antoniades, C. Shirodaria, T. Van Assche, C. Cunnington, I. Tegeder, J. Lötsch,
T.J. Guzik, P. Leeson, J. Diesch, D. Tousoulis, C. Stefanadis, M. Costigan, C.J.
Woolf, N.J. Alp, K.M. Channon, GCH1 haplotype determines vascular and plasma
biopterin availability in coronary artery disease effects on vascular superoxide
production and endothelial function, J. Am. Coll. Cardiol. 52 (2008) 158–165.
[3] J. Audrain-McGovern, D. Rodriguez, V. Patel, M.S. Faith, K. Rodgers, J. Cuevas,
15 How do psychological factors influence adolescent smoking progression? The
evidence for indirect effects through tobacco advertising receptivity, Pediatrics
: Males 117 (2006) 1216–1225.
: Females [4] C.R. Cloninger, T.R. Przybeck, D.M. Svrakic, R.D. Wetzel, The Temperament and
Character Inventory (TCI): A Guide to its Development and Use, Center for
Psychobiology of Personality, Washington University, St Louis, MI, 1994.
10 [5] C.R. Cloninger, D.M. Svrakic, T.R. Przybeck, Can personality assessment predict
C/C C/T T/T C/C C/T T/T future depression? A twelve-month follow-up of 631 subjects, J. Affect. Disord.
92 (2006) 35–44.
GCH1 genotypes [6] J.R. Cooper, F.E. Bloom, R.H. Roth, The Biochemical Basis of Neurophamacology,
8th ed., Oxford University Press, Oxford, 2003.
Fig. 2. Post-hoc analysis of the effects of the GCH1 genotypes on the scores of novelty [7] P.T. Costa Jr., R.R. McCrae, The NEO-PI/NEO-FFI manual supplement, Odessa,
seeking in males and females. Error bars indicate SD of the mean. Psychol. Assess. Resour. (1989).
 R. Sadahiro et al. / Neuroscience Letters 503 (2011) 220–223
[8] A. Doehring, C. Antoniades, K.M. Channon, I. Tegeder, J. Lotsch, Clinical genetics
of functionally mild non-coding GTP cyclohydrolase 1 (GCH1) polymorphisms
modulating pain and cardiovascular risk, Mutat. Res. 659 (2008) 195–201.
[9] R.F. Farmer, J.R. Seeley, Temperament and character predictors of depressed
mood over a 4-year interval, Depress. Anxiety 26 (2009) 371–381.
[10] N. Gordon, Segawa’s disease: dopa-responsive dystonia, Int. J. Clin. Pract. 62
(2008) 943–946.
[11] K. Hyland, R.S. Gunasekara, T.L. Munk-Martin, L.A. Arnold, T. Engle, The hph-1
mouse: a model for dominantly inherited GTP-cyclohydrolase deficiency, Ann.
Neurol. 54 (suppl 6) (2003) 46–48.
[12] N. Kijima, R. Saito, M. Suzuki, A. Yoshino, Y. Ono, M. Kato, T. Kitamura,
Cloninger’s seven factor model of temperament and character and Japanese
version of Temperament and Character Inventory (TCI), Jpn. J. Psychiatr. Diagn.
7 (1996) 379–399 (Japanese).
[13] A.A. Lam, K. Hyland, S.J. Heales, Tetrahydrobiopterin availability, nitric oxide
metabolism and glutathione status in the hph-1 mouse; implications for the
pathogenesis and treatment of tetrahydrobiopterin deficiency states, J. Inherit.
Metab. Dis. 30 (2007) 256–262.
[14] Y.F. Liao, T.S. Zeng, L.L. Chen, Y.M. Li, F. Yu, L.J. Hu, L. Yue, Association of a
functional polymorphism (C59038T) in GTP cyclohydrolase 1 gene and type 2
diabetic macrovascular disease in the Chinese population, J. Diabetes Compli-
cations 24 (2010) 313–319.
[15] A. Reif, E. Kiive, T. Kurrikoff, M. Paaver, S. Herterich, K. Konstabel, T. Tulviste,
K.P. Lesch, J. Harro, A functional NOS1 promoter polymorphism interacts with
adverse environment on functional and dysfunctional impulsivity, Psychophar-
macology 214 (2011) 239–248.
[16] R. Sadahiro, A. Suzuki, N. Shibuya, M. Kamata, Y. Matsumoto, K. Goto, K. Otani,
Association study between a functional polymorphism of tyrosine hydroxylase
223
gene promoter and personality traits in healthy subjects, Behav. Brain Res. 17
(2010) 209–212.
[17] L.I. Serova, M. Filipenko, N. Schilt, M. Veerasirikul, E.L. Sabban, Estrogen-
triggered activation of GTP cyclohydrolase 1 gene expression: role of estrogen
receptor subtypes and interaction with cyclic AMP, Neuroscience 140 (2006)
1253–1263.
[18] M. Shimoji, K. Hirayama, K. Hyland, G. Kapatos, GTP cyclohydrolase I gene
expression in the brains of male and female hph-1 mice, J. Neurochem. 72
(1999) 757–764.
[19] H. Shiraishi, A. Suzuki, T. Fukasawa, T. Aoshima, Y. Ujiie, G. Ishii, K. Otani,
Monoamine oxidase A gene promoter polymorphism affects novelty seeking
and reward dependence in healthy study participants, Psychiatr. Genet. 16
(2006) 55–58.
[20] I. Tegeder, M. Costigan, R.S. Griffin, A. Abele, I. Belfer, H. Schmidt, C. Ehnert, J.
Nejim, C. Marian, J. Scholz, T. Wu, A. Allchorne, L. Diatchenko, A.M. Binshtok, D.
Goldman, J. Adolph, S. Sama, S.J. Atlas, W.A. Carlezon, A. Parsegian, J. Lötsch, R.B.
Fillingim, W. Maixner, G. Geisslinger, M.B. Max, C.J. Woolf, GTP cyclohydrolase
and tetrahydrobiopterin regulate pain sensitivity and persistence, Nat. Med. 12
(2006) 1269–1277.
[21] G. Wegener, V. Volke, R. Rosenberg, Endogenous nitric oxide decreases hip-
pocampal levels of serotonin and dopamine in vivo, Br. J. Pharmacol. 130 (2000)
575–580.
[22] L. Zhang, F. Rao, K. Zhang, S. Khandrika, M. Das, S.M. Vaingankar, X. Bao,
B.K. Rana, D.W. Smith, J. Wessel, R.M. Salem, J.L. Rodriguez-Flores, S.K.
Mahata, N.J. Schork, M.G. Ziegler, D.T. O’Connor, Discovery of common human
genetic variants of GTP cyclohydrolase 1 (GCH1) governing nitric oxide, auto-
nomic activity, and cardiovascular risk, J. Clin. Invest. 117 (2007) 2658–
2671.