Genes, Brain and Behavior (2008) 7: 698704

# 2008 The Authors

Journal compilation # 2008 Blackwell Publishing Ltd

Association between mitochondrial DNA 10398A>G

polymorphism and the volume of amygdala
H. Yamasue*,,, C. Kakiuchi, M. Tochigi,,
H. Inoue, M. Suga, O. Abe, H. Yamada,
T. Sasaki, M. A. Rogers, S. Aoki, T. Kato
and K. Kasai

Department of Neuropsychiatry, Graduate School of Medicine,

University of Tokyo, Tokyo, Laboratory for Molecular Dynamics
of Mental Disorders, Brain Science Institute, RIKEN, Wako, and

Department of Radiology, Graduate School of Medicine, University of Tokyo, Tokyo, Japan

*Corresponding author: Hidenori Yamasue, Department of
Neuropsychiatry, Graduate School of Medicine, University of
Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan.
E-mail: yamasue-tky@umin.ac.jp

Mitochondrial calcium regulation plays a number of

important roles in neurons. Mitochondrial DNA (mtDNA)
is highly polymorphic, and its interindividual variation is
associated with various neuropsychiatric diseases and
mental functions. An mtDNA polymorphism, 10398A>G,
was reported to affect mitochondrial calcium regulation.
Volume of hippocampus and amygdala is reportedly
associated with various mental disorders and mental
functions and is regarded as an endophenotype of mental
disorders. The present study investigated the relationship
between the mtDNA 10398A>G polymorphism and the
volume of hippocampus and amygdala in 118 righthanded healthy subjects. The brain morphometry using
magnetic resonance images employed both manual tracing volumetry in the native space and voxel-based morphometry (VBM) in the spatially normalized space.
Amygdala volume was found to be significantly larger in
healthy subjects with 10398A than in those with 10398G
by manual tracing, which was confirmed by the VBM.
Brain volumes in the other gray matter regions and all
white matter regions showed no significant differences
associated with the polymorphism. These provocative
findings might provide a clue to the complex relationship
between mtDNA, brain structure and mental disorders.
Keywords: Amygdala, bipolar disorder, endophenotype, mitochondria, MRI, volumetry, voxel-based morphometry
Received 29 December 2007, revised 28 April 2008, accepted
for publication 28 April 2008

In addition to their fundamental function in energy production,

mitochondria play a role in calcium signaling (Campanella et al.

698

2004). In neurons, mitochondria distribute in dendrites and

growth cones (Li et al. 2004) and mitochondrial calcium
regulation has significant influences on a wide variety of
neuronal functions such as exocytosis (Rizzuto 2003), synaptic plasticity (Rizzuto 2001) and apoptosis (Duchen 2004).
Mitochondrial DNA (mtDNA) is highly polymorphic in comparison with nuclear genome, and interindividual variations of
mtDNA have been reported to be associated with human
health and diseases. Mitochondrial DNA polymorphisms are
reportedly associated with human functional variability such
as cognitive function (Skuder et al. 1995), personality (Kato
et al. 2004) and longevity (Tanaka et al. 2000). In addition,
mtDNA polymorphisms have been reported to be associated
with several neuropsychiatric diseases such as Alzheimers
disease, Parkinsons disease (reviewed in Howell et al. 2005),
schizophrenia and bipolar disorder (reviewed in Kato 2001).
In spite of putative roles of mtDNA in human health and
diseases, none of the association finding is conclusive
because the functional consequence of mtDNA polymorphisms has not been well studied and the association studies
of complex diseases tend to report false-positive findings
(Sullivan 2007).
With regard to the functional consequence of mtDNA
polymorphisms, a recent paper using updated calcium imaging technologies and extensive whole mtDNA sequencing
identified two closely linked mtDNA polymorphisms affecting
mitochondrial calcium level and pH. Among two closely linked
mtDNA polymorphisms, 10398A>G was regarded as having
functional significance (Kazuno et al. 2006). Although mtDNA
10398A was reportedly associated with Alzheimers disease
(van der Walt et al. 2004), Parkinsons disease (van der Walt
et al. 2003) and bipolar disorder (Kato et al. 2001; McMahon
et al. 2000), none of the finding is consistent across studies
(Kirk et al. 1999; Raule et al. 2007). One way to overcome
such inconsistency in association findings of complex neuropsychiatric diseases is to use an endophenotype, a measurable biological parameter that is determined by genetic
factors and associated with the diseases (Gottesman & Gould
2003).
Neuroanatomy measured by magnetic resonance imaging
(MRI) is one of the most promising of endophenotype
candidates. In particular, reduced volume of hippocampus
or altered volume of amygdala has been reported in various
mental disorders such as schizophrenia, major depression,
bipolar disorder and posttraumatic stress disorder (Geuze
et al. 2005; Honea et al. 2005; Strakowski et al. 2002). The
volume of these structures has also been reported to be
associated with interindividual variation of mental functions
such as intelligence (Amat et al. 2008) and personality (Omura
et al. 2005; Yamasue et al. 2007b). Previous studies have
suggested that amygdala and hippocampus are vulnerable to
doi: 10.1111/j.1601-183X.2008.00408.x

Mitochondrial DNA and brain volume

the stress at cellular level (Kubota et al. 2006; Tsuchiya et al.

1999), which might indirectly suggest these structures would
be especially vulnerable to mtDNA variation. For example, the
neurochemical change in hippocampus of animal with mtDNA
defects has been reported in a recent study (Kubota et al. 2006).
In addition, an autopsy case of MELAS showed small necrotic
foci in amygdala and hippocampus (Tsuchiya et al. 1999).
Taken together, an association analysis between the genotype and the endophenotype, functional mtDNA polymorphism and hippocampal and amygdala volumes in this case
might be a promising strategy.
In this study, the effects of mtDNA 10398A>G polymorphism on the volume of hippocampus and amygdala were
investigated in healthy humans using a combination of
manual tracing and voxel-based morphometry (VBM). Based
on the previous literature on bipolar disorder, in which
association with mtDNA 10398A and larger volume of amygdala and smaller volume of hippocampus were reported
(reviewed in Beyer & Krishnan 2002; Hajek et al. 2005), we
predicted that 10398A might be associated with larger
amygdala and smaller hippocampus.

Materials and methods

Participants
Participants included 118 right-handed Japanese individuals (80
males/38 females), mainly college students, hospital staff and their
acquaintances. To minimize the effects of aging on brain morphology,
the age range of the participants was restricted from 21 to 55 years.
The participants were interviewed by a trained psychiatrist (H.Y. or
M.S.) to be screened for the presence or absence of neuropsychiatric
disorders through the Structured Clinical Interview for DSM-IV Axis I
Disorder, Non-patient Edition (SCID-NP) (First et al. 1997). These
interviews were performed on the same day as magnetic resonance
scanning.
Socioeconomic status (SES) and parental SES were assessed using
the Hollingshead scale (Hollingshead 1965). Handedness was assessed based on the Edinburgh Inventory (Oldfield 1971).
The exclusion criteria were neurological illness, traumatic brain
injury with any known cognitive consequences or loss of consciousness for more than 5 min and a history of psychiatric disease in
themselves or a family history of axis I disorder in their first-degree
relatives. The Ethics Committee of the University of Tokyo Hospital
approved this study. After a complete explanation of the study to the
participants, written informed consent was obtained.

Genotyping
Blood samples were collected from all individuals, and DNA was
extracted by standard methods. The mtDNA 10398A>G and
10410T>C polymorphisms were genotyped by sequencing. The
mt10410 polymorphism, which does not have neuropsychiatric
associations, was genotyped to test the specificity of association
between mt10398 polymorphism and regional brain volume.

MRI acquisition
The method of 1.5-mm-slice, high-spatial-resolution MRI acquisition was
the same as those of our previous studies (Yamasue et al. 2003, 2004,
2007a). Briefly, the MRI data were obtained using a 1.5-Tesla scanner
(General Electric Signa Horizon Lx version 8.2; GE Medical Systems,
Milwaukee, WI, USA). Three-dimensional Fourier transform spoiled
gradient recalled acquisition with steady state was used because it
affords excellent contrast between the gray matter and the white matter
Genes, Brain and Behavior (2008) 7: 698704

in the evaluation of brain structures. The repetition time was 35 milliseconds, the echo time 7 milliseconds with one repetition, the nutation
angle 30 degrees, the field of view 24 cm and the matrix 256
256
(192)
124. A trained neuroradiologist (H.Y. or O.A.) evaluated the MRI
scans and found no gross abnormalities in any of the participants.

Manual tracing for hippocampus and amygdala

volumetry
The amygdala and hippocampus gray matter regions of interest (ROIs)
were outlined manually by one rater (H.I.) using a software package
for medical image analysis (3D SLICER 1.2.2, software available at http://
www.slicer.org) without knowledge of group status or genotype. By
applying the software, we can view under tracing ROI in orthogonal
slices and edited in every slice. The landmarks to delineate the ROIs in
the present study were refined from those in our previous studies
(Kasai et al. 2004; Yamasue et al. 2004). As described in detail below,
to accurately measure the volume of these structures in the present
study, we developed an additional protocol for delineating the anterior
boundary of amygdala and boundary between amygdala and hippocampus similar to those described in previous literature (Schumann
et al. 2004; Van Erp et al. 2002).
Dentate gyrus, CA fields, subiculum, presubiculum and parasubiculum were referred to as the hippocampus, while the fornix, fimbria
and alveus were not included in the volumetric measurements. The
tracing of hippocampus was mainly performed in the sagittal plane,
although we found it essential to trace on axial and coronal planes for
cases in which the borders were ambiguous on sagittal slices. Tracing
began on the slice in the most lateral extent of the ventricular temporal horn on which the hippocampus was first visible. The outline
represents the inferior border (determined by drawing a line through
the white matter separating the hippocampus from the parahippocampal and fusiform gyri), superior border (determined by drawing
a line through the alveus separating the hippocampus from the lateral
ventricles) and the posterior border (determined by following the
structure to its furthest posterior extent). More medially, the anterior
hippocampus was separated from amygdala by a thin line of white
matter between the two structures (alveus). Once drawn, hippocampal ROIs could be viewed in any plane and as a three-dimensional
object, for any further editing.
For the tracing of amygdala boundaries, the initial tracing process
involved defining the borders in coronal sections starting with the
most caudal level in which the amygdala was visible. At its caudal
extent, the amygdala is bordered dorsally by the substantia innominata, laterally by the putamen and ventrally by the temporal horn of
the lateral ventricle. The medial surface of the amygdala abuts the
optic tract. Proceeding rostrally, it is bordered dorsally by fibers of the
anterior commissure as well as the substantia innominata. The lateral
border is formed by white matter of the temporal lobe. The ventral
surface is formed by the temporal horn of the lateral ventricle.
However, because the hippocampus often appears to be fused with
the ventral surface of the amygdala, a more reliable boundary is the
alveus, the white matter that forms the dorsal surface of the
hippocampus. In more rostral sections, the hippocampus decreases
in size and the entorhinal cortex begins to form part of the medial
surface of the amygdala. At this point, a thin band of white matter
separates the amygdala from the entorhinal cortex. In most rostral
sections, the dorsomedial surface of the amygdala forms a portion of
the medial surface of the brain. The amygdala is bordered laterally by
white matter of the temporal lobe, ventrally by the temporal horn of
the lateral ventricle and by subamygdaloid white matter and ventromedially by the entorhinal cortex. At the rostral pole of the amygdala,
the outlining rules are very similar to what has just been described
above. However, the gray matterwhite matter boundaries are more
difficult to delineate. Therefore, it was necessary to confirm the
rostral boundary of the amygdala by reviewing the outlines in sagittal
images.
For inter-rater reliability, two raters (H.I. and M.A.R.) blind to group
membership, independently drew ROIs. Ten cases were selected at
random, and the raters drew ROIs on every slice. The intraclass
correlation coefficient was 0.87/0.85 for left/right amygdala and 0.93/
0.94 for left/right hippocampus. Intra-rater reliability, computed

699

Yamasue et al.
by using all the slices from one randomly selected brain and measured
by one rater (H.I.) at two separate times (approximately 2 months
apart), was >0.95 for all structures.
Intracranial content (ICC), total gray matter, white matter and
cerebrospinal fluid volumes were calculated from the VBM procedure
using SPM2 (Good et al. 2001). Then, ICC was calculated by summing
up total gray matter, white matter and cerebrospinal fluid volume. To
validate this method, the ICCs of an independent sample of 50 adult
participants were measured by both the VBM and the intensity-based
semiautomated segmentation procedure using ANALYZE PC 3.0 (Yamasue et al. 2003). We then confirmed that the calculated intraclass
correlation coefficient for ICCs was adequate (0.96).

adopting relative volume as the dependent variable, genotype as

the between-subject factor and hemisphere as the within-subject
factor, separately for amygdala and hippocampus. Individual difference in ICC is considered to be one of the major contributors to
individual differences in regional brain volume. Therefore, relative
volumes [(1000
absolute ROI volume)/ICC] were used as the
dependent variable. Of note, the statistical conclusions reported
below remained the same when analysis of covariance (ANCOVA) with
absolute volume as the dependent variable and ICC as the covariate
were used. Statistically significant level was set at P < 0.05.
For testing regional specificity, volumes of total gray matter, total
white matter, CSF and ICC were also compared between genotypes
using independent t-tests.

Image processing for VBM

Statistical analysis of VBM

Image processing for VBM (Ashburner & Friston 2000; Good et al.
2001), a fully automatic technique for computational analysis of
differences in local brain tissue volume throughout the entire brain,
was conducted using SPM 2 (Institute of Neurology, London, UK). This
method, also applied in our recent study (Yamasue et al. 2007a),
involves the following steps: (1) spatial normalization of all images to
a standardized anatomical space by removing differences in overall
size, position and global shape; (2) extraction of gray and white matter
from the normalized images and (3) analysis of differences in local
gray and white matter volume across the whole brain (Ashburner &
Friston 2000). The spatial normalization to the standard anatomical
space was performed in a two-stage process. In the first step, each
image was registered to the International Consortium for Brain
Mapping template (Montreal Neurological Institute, Montreal, Canada),
which approximates Talairach space. This step applied a 12-parameter
affine transformation to correct for image size and position. Regional
volumes were preserved while corrections for global differences in
whole brain volume were made. The normalized images of all
participants were averaged and smoothed with a Gaussian kernel of
8-mm full width at half maximum (FWHM) and then used as a new
template with reduced scanner- and population-specific bias. In the
second normalization step, we locally deformed each image of our
entire group to the new study-specific template using a nonlinear
spatial transformation. This accounts for the remaining shape differences between the images and the template and improves the
overlap of corresponding anatomical structures. Finally, using a modified mixture model cluster analysis, normalized images were corrected for nonuniformities in signal intensity and partitioned using
study-specific customized prior probability map into gray and white
matter, cerebrospinal fluid (CSF) and background. To remove unconnected nonbrain voxels (e.g. rims between brain surface and meninges), a series of morphological erosions and dilations to the
segmented images were applied. In an intensity modulation step,
voxel values of the segmented images were multiplied by the
measure of warped and unwarped structures derived from the
nonlinear step of the spatial normalization (Jacobian determinant).
This step converts relative regional gray matter density to absolute
gray matter density expressed as the amount of gray matter per unit
volume of brain tissue prior to spatial normalization. The resulting
modulated gray matter images were smoothed with a Gaussian
kernel of 12-mm FWHM.

Statistical analyses
Group comparison for demographic and clinical variables
and 10398A>G frequencies
t-Tests were performed to assess group differences in age, handedness, self SES and parental SES between participants with 10398A
and 10398G. Chi-squared tests were employed to test group differences in sex ratio of 10398A>G groups.

Effects of 10398A>G genotype on manually traced

volume
The effects of 10398A>G genotype on manually traced volumes were
assessed using repeated measures analysis of variance (ANOVA)

700

Statistical analyses of VBM were performed using an ANCOVA model

(Friston et al. 1990). To account for global anatomical variations, the
ICC was treated as a confounding covariate. To detect the neuroanatomical correlates of 10398A>G genotype, statistical analysis treated ICC as confounding covariate and 10398A>G genotype as
condition. To test hypotheses with respect to regionally specific
association with 10398A>G genotype, the estimates were compared
using two linear contrasts. The resulting set of voxel values for each
contrast constituted a statistical parametric map of the t-statistic
(SPM{t }). The SPM{t }s were displayed at an uncorrected threshold of
P < 0.005 for graphical reporting. We only discuss results in the text
and in tables that survive a correction at 0.05 for the search volumes.
The statistics in the tables are transformed to a z score to make them
more intuitive. The significance of each region was corrected for
multiple comparisons using false discovery rate (FDR) (Genovese
et al. 2002). The statistical significance level was set at FDR-corrected
P < 0.05. While significant effects were explored throughout the
entire gray matter regions, small volume correction was employed in
hippocampus and amygdala based on previous literature (Beyer &
Krishnan 2002; Hajek et al. 2005). In contrast to the whole gray matter
exploration, family-wise error (FWE)-corrected P was conservatively
employed to detect findings within the searched volumes in the
hypothesized regions. The searched volumes were defined using
WFU_PICKATLAS (a software available at http://fmri.wfubmc.edu/cms/
software#PickAtlas).

Results
Demographic and clinical variables and 10398A>G
No significant group difference between 10398A>G polymorphism was found in age, handedness, self SES, parental
SES or sex ratio (P > 0.36).

Effects of 10398A>G genotype on manually traced

volumetric measures
The repeated measures ANOVA showed a significant
10398A>G genotype effect on amygdala volume
(F1,116 4.77, P 0.031), while the interaction between
genotype and hemisphere on amygdala volume was not
significant (F1,116 0.28, P 0.60). These results indicated
that amygdala (left right) volume was significantly larger in
participants with 10398A than in those with 10398G (effect
size: total, 0.44; left, 0.36; right, 0.42) (% increase: total,
5.2%; left, 4.7%; right, 5.6%) (Fig. 1, Table 1). The
10398A>G genotype effect on manually traced amygdala
volume was preserved after controlling age (F1,115 4.75,
P 0.031), sex (F1,115 3.66, P 0.058) and both effects
(F1,114 3.63, P 0.059). In contrast, there was no significant genotype main effect (F1,116 0.83, P 0.37) (effect
Genes, Brain and Behavior (2008) 7: 698704

Mitochondrial DNA and brain volume

size: total, 0.17; left, 0.17; right, 0.16) or interaction

(F1,116 0.0, P 0.98) between genotype and hemisphere
for hippocampus volume (Table 1).
Total gray matter, total white matter, CSF and ICC volumes
were not significantly different between genotypes (P >
0.22) (Table 1).

Effects of 10398A>G genotype on regional brain

volume analyzed by VBM

Figure 1: Manually traced amygdala volume and 10398A>G

genotype. Means and distributions of left and right hemisphere
for relative manually traced amygdala volume [(absolute volume

1000)/ICC] in which individuals with 10398A genotype exhibited
bilateral enlargement compared with individuals with 10398G
genotype (F1,116 4.77, P 0.031). Means are represented by
solid horizontal lines drawn on each groups distribution.

The VBM showed significantly larger regional gray matter

volume in left amygdala (peak co-ordinate [26, 2, 14],
z 3.01, FWE-corrected P 0.01 with 1.76 ml searched
volume, cluster size 880 mm3) of individuals with 10398A
than in those with 10398G after correction for multiple
comparisons. The additional VBM analyses also revealed
significantly preserved 10398A>G genotype effect on
regional volume in left amygdala with age ([26, 4, 14],
z 3.36, FWE-corrected P 0.004), sex ([26, 4, 14],
z 3.32, FWE-corrected P 0.004) and both ([26, 4,
14], z 3.32, FWE-corrected P 0.004) as confounding
covariate. Regional brain volumes in other gray matter regions

Table 1: Subject characteristics and volumetric measures

10398A (n 35)
Variable
Demographic variables
Male/female
Age (range)
Handedness (range)*
SES
Parental SES
Volumetric measures
Absolute volume (ml)
Total gray matter
Total white matter
CSF
ICC
Left amygdala
Right amygdala
Left hippocampus
Right hippocampus
Relative volume (0/00)
Left amygdala
Right amygdala
Left hippocampus
Right hippocampus

Mean

21/14
30.7 (2255)
96.3 (64100)
1.6
2.1

10398G (n 83)
SD

8.0
7.4
0.6
0.7

Mean

59/24
30.5 (2154)
97.8 (35100)
1.6
2.2

SD

6.7
7.8
0.7
0.8

741.5
446.5
369.9
1557.9
1.55
1.63
2.73
2.79

81.4
50.8
67.3
173.7
0.24
0.23
0.40
0.44

756.8
458.1
373.9
1588.8
1.51
1.57
2.70
2.75

68.7
45.6
62.5
154.5
0.22
0.22
0.39
0.41

1.00
1.05
1.76
1.79

0.13
0.13
0.22
0.20

0.95
0.99
1.71
1.74

0.12
0.12
0.27
0.28

Group comparison

w2 1.39, P
t(116) 0.15, P
t(107) 0.92, P
t(116) 0.17, P
t(112) 0.58, P

t(116)
t(116)
t(116)
t(116)

1.05, P
1.22, P
0.31, P
0.96, P

0.24
0.88
0.36
0.87
0.56

0.30
0.22
0.76
0.34

F1,116 4.77, P 0.031

F1,116 0.83, P 0.37

*Determined using Edinburgh Inventory (Oldfield 1971): scores greater than 0 indicate right-handedness. A score of 100 indicates strong righthandedness.

Assessed using the Hollingshead scale (Hollingshead 1965). Higher scores indicate lower educational and/or occupational status.

Degrees of freedom varied because of unavailability of data on some subjects.

Significant main effect of A10398G genotype was found, although the interaction between genotype and hemisphere was not significant
(F1,116 0.28, P 0.60). These results indicate total amygdala (left right) enlargement in subjects with 10398A than in those with 10398G.
Genes, Brain and Behavior (2008) 7: 698704

701

Yamasue et al.

and all white matter regions did not reach statistical significance after correction for multiple comparisons (Fig. 2).

Effects of 10410T>C genotype on regional brain

volume
To examine the specificity of association between amygdala
and mt10398T>C polymorphism, the association between
amygdala volume and mt10410 polymorphism was examined
using both the manual tracing and the VBM. Both the
methodologies showed no significant effect of mt10410
polymorphism on the amygdala volume (manually traced
volume: F 0.72, P 0.398; VBM: no suprathreshold voxel
even at very liberal threshold, uncorrected P > 0.05).

Discussion
The present study revealed significantly larger amygdala
volume in healthy subjects with 10398A than in those with
10398G, which was confirmed by two independent methodologies, VBM and manual tracing.
Increased amygdala volume has been reported in several
mental diseases such as borderline personality disorder
(Minzenberg et al. 2008) and bipolar disorder (Altshuler
et al. 2007; Strakowski et al. 1999), while reduced amygdala
volume has been reported in major depression (Strakowski

et al. 2002), schizophrenia (Honea et al. 2005), dissociative

identity disorder (Vermetten et al. 2006) and conduct disorder
(Sterzer et al. 2007). Larger amygdala volume was also
associated with lower scores of neuroticism (Omura et al.
2005). All these findings are still not conclusive, and the
results are conflicting. The reason for such inconsistency not
yet understood. Amygdala volume may be related to
response to stress or early adversity rather than being directly
related to diagnostic category (Bremner 2006) or may be
affected by other behavioral factors such as aggression
(Krishnan 1999).
The different methods used to measure amygdala volume
among previous studies, such as image resolution, number of
slices measured and the definition of amygdala, might also
contribute to the inconsistency (Brierley et al. 2002). Although
many recent studies used only coronal slices to trace amygdala, such approaches are not optimal, particularly for the
anterior boundary of the amygdala and the boundary between
the amygdala and the hippocampus, which are difficult to
discern accurately from coronal slices. In the present study,
a more accurate tracing method was used, in which every
slice was checked in each of three orthogonal planes.
The 10398A>G polymorphism has been reported to be
associated with human health and disease. The 10398A is
reportedly associated with neuropsychiatric disorders such as
Alzheimers disease (van der Walt et al. 2004), Parkinsons
disease (van der Walt et al. 2003) and bipolar disorder (Kato

Figure 2: Regional brain volume revealed by VBM and 10398A>G genotype.

The gray matter regions that showed
associations with 10398A>G genotype
are rendered in the Montreal Neurological
Institute (MNI) space. Left: Statistical
parametric map in the three orthogonal
projections
shows
voxels
where
10398A>G polymorphism was associated with regional gray matter volume
(voxel threshold: uncorrected P <
0.005). Right: After FWE correction for
multiple comparisons, those gray matter
regions showing statistically significant
associations with 10398A>G genotype
were rendered onto the averaged images
of the whole sample (N 118) (voxel
threshold: FWE-corrected P < 0.05). Left
amygdala (peak co-ordinate [26, 2,
14], z 3.01, FWE-corrected P 0.01
with 1.76 ml searched volume, cluster
size 880 mm3).

702

Genes, Brain and Behavior (2008) 7: 698704

Mitochondrial DNA and brain volume

et al. 2001; McMahon et al. 2000), as well as various cancers

(Canter et al. 2005; Darvishi et al. 2007). The 10398G was
reported to be associated with longevity (Niemi et al. 2005).
However, all these associations are still controversial. The
10398 A/G characterizes the European haplogroups I, J and K
(van der Walt et al. 2004) and Asian-specific super-haplogroup
M (Sudoyo et al. 2002). The majority of Europeans have
10398A, while 10398G is more frequent than 10398A in
Asians (Kazuno et al. 2006).
As summarized above, both amygdala volume and the
10398A>G polymorphisms are reportedly associated with
various neuropsychiatric phenotypes, but there has been no
study to test the relationship between amygdala volume and
mtDNA 10398A>G polymorphism. Because mtDNA
10398A>G polymorphism alters mitochondrial calcium levels
and mitochondrial calcium regulation plays a role in neuroplasticity and apoptosis, mtDNA 10398A>G polymorphism
might also affect the response of neurons to neuroplastic
changes or vulnerability to cellular stress. It has been hypothesized that stress affects neurons in cerebral cortex differently
from those in the amygdala, as suggested by findings in
patients with post-traumatic stress disorder (PTSD) (Bremner
2006) and animal experiments (Mitra et al. 2005; Radley et al.
2006; Vyas et al. 2002). Taken together, we can hypothesize
that mtDNA 10398A>G polymorphism might have caused an
enlargement of amygdala through the modulation of stressinduced synaptic plasticity, although such a preliminary hypothesis should be confirmed in a large prospective study. Replication in populations of different ethnicity will be important
before drawing conclusions.
Recently developed voxel-by-voxel computational morphometry has become the gold standard to comprehensively
explore brain structural correlates of a certain phenotype
(Ashburner & Friston 2000). However, it has been argued
that computational morphometry should be used with caution
when examining anatomically localized structures because
the spatial normalization process could reduce information of
individual difference in brain structure, especially in highly
localized structures (Bookstein 2001). Therefore, the advantage of manually traced volumetry has recently been recognized for examination of small brain structures such as
hippocampus and amygdala. It has been suggested that
manually traced volumetry and VBM would provide different
aspects of information and should thus be used in tandem
(Giuliani et al. 2005).

References
Altshuler, L.L., Bartzokis, G., Grieder, T., Curran, J. & Mintz, J. (2007)
Amygdala enlargement in bipolar disorder and hippocampal reduction in schizophrenia: an MRI study demonstrating neuroanatomic
specificity. Arch Gen Psychiatry 55, 663664.
Amat, J.A., Bansal, R., Whiteman, R., Haggerty, R., Royal, J. &
Peterson, B.S. (2008) Correlates of intellectual ability with morphology of the hippocampus and amygdala in healthy adults. Brain
Cogn 66, 105114.
Ashburner, J. & Friston, K.J. (2000) Voxel-based morphometry: the
methods. Neuroimage 11, 805821.
Beyer, J.L. & Krishnan, K.R. (2002) Volumetric brain imaging findings
in mood disorders. Bipolar Disord 4, 89104.
Bookstein, F.L. (2001) Voxel-based morphometry should not be used
with imperfectly registered images. Neuroimage 14, 14541462.
Genes, Brain and Behavior (2008) 7: 698704

Bremner, J.D. (2006) Traumatic stress: effects on the brain. Dialogues

Clin Neurosci 8, 445461.
Brierley, B., Shaw, P. & David, A.S. (2002) The human amygdala:
a systematic review and meta-analysis of volumetric magnetic
resonance imaging. Brain Res Rev 39, 84105.
Campanella, M., Pinton, P. & Rizzuto, R. (2004) Mitochondrial Ca2
homeostasis in health and disease. Biol Res 37, 653660.
Canter, J.A., Kallianpur, A.R., Parl, F.F. & Millikan, R.C. (2005)
Mitochondrial DNA G10398A polymorphism and invasive breast
cancer in African-American women. Cancer Res 65, 80288033.
Darvishi, K., Sharma, S., Bhat, A.K., Rai, E. & Bamezai, R.N. (2007)
Mitochondrial DNA G10398A polymorphism imparts maternal
Haplogroup N a risk for breast and esophageal cancer. Cancer Lett
249, 249255.
Duchen, M.R. (2004) Roles of mitochondria in health and disease.
Diabetes 53, S96S102.
First, M.B., Spitzer, R.L., Gibbon, M. & Williams, J.B.W. (1997)
Structured Clinical Interview for DSM-IV Axis I Disorders: Nonpatient Edition. Biometrics Research Department. New York State
Psychiatric Institute, New York. (Japanese translation: Kitamura, T.
& Okano, T. (2003) Nihon Hyoron-sha publishers, Tokyo).
Friston, K.J., Frith, C.D., Liddle, P.F., Dolan, R.J., Lammertsma, A.A.
& Frackowiak, R.S. (1990) The relationship between global and local
changes in PET scans. J Cereb Blood Flow Metab 10, 458466.
Genovese, C.R., Lazar, N.A. & Nichols, T. (2002) Thresholding of
statistical maps in functional neuroimaging using the false discovery rate. Neuroimage 15, 870878.
Geuze, E., Vermetten, E. & Bremner, J.D. (2005) MR-based in vivo
hippocampal volumetrics: 2. Findings in neuropsychiatric disorders.
Mol Psychiatry 10, 160184.
Giuliani, N.R., Calhoun, V.D., Pearlson, G.D., Francis, A. & Buchanan,
R.W. (2005) Voxel-based morphometry versus region of interest:
a comparison of two methods for analyzing gray matter differences
in schizophrenia. Schizophr Res 74, 135147.
Good, C.D., Johnsrude, I., Ashburner, J., Henson, R.N., Friston, K.J. &
Frackowiak, R.S. (2001) Cerebral asymmetry and the effects of sex
and handedness on brain structure: a voxel-based morphometric
analysis of 465 normal adult human brains. Neuroimage 14,
685700.
Gottesman, I.I. & Gould, T.D. (2003) The endophenotype concept in
psychiatry: etymology and strategic intentions. Am J Psychiatry
160, 636645.
Hajek, T., Carrey, N. & Alda, M. (2005) Neuroanatomical abnormalities
as risk factors for bipolar disorder. Bipolar Disord 7, 393403.
Hollingshead, A.B. (1965) Two-factor Index of Social Position. Yale
University Press, New Haven, CT.
Honea, R., Crow, T.J., Passingham, D. & Mackay, C.E. (2005)
Regional deficits in brain volume in schizophrenia: a meta-analysis of
voxel-based morphometry studies. Am J Psychiatry 162, 22332245.
Howell, N., Elson, J.L., Chinnery, P.F. & Turnbull, D.M. (2005) mtDNA
mutations and common neurodegenerative disorders. Trends
Genet 21, 583586.
Kasai, K., McCarley, R.W., Salisbury, D.F., Onitsuka, T., Demeo, S.,
Yurgelun-Todd, D., Kikinis, R., Jolesz, F.A. & Shenton, M.E. (2004)
Cavum septi pellucidi in first-episode schizophrenia and first-episode affective psychosis: an MRI study. Schizophr Res 71, 6576.
Kato, T. (2001) The other, forgotten genome: mitochondrial DNA and
mental disorders. Mol Psychiatry 6, 625633.
Kato, T., Kunugi, H., Nanko, S. & Kato, N. (2001) Mitochondrial DNA
polymorphisms in bipolar disorder. J Affect Disord 62, 151164.
Kato, C., Umekage, T., Tochigi, M., Otowa, T., Hibino, H., Ohtani, T.,
Kohda, K., Kato, N. & Sasaki, T. (2004) Mitochondrial DNA polymorphisms and extraversion. Am J Med Genet B Neuropsychiatr
Genet 128, 7679.
Kazuno, A.A., Munakata, K., Nagai, T., Shimozono, S., Tanaka, M.,
Yoneda, M., Kato, N., Miyawaki, A. & Kato, T. (2006) Identification
of mitochondrial DNA polymorphisms that alter mitochondrial
matrix pH and intracellular calcium dynamics. PLoS Genet 2, e128.
Kirk, R., Furlong, R.A., Amos, W., Cooper, G., Rubinsztein, J.S.,
Walsh, C., Paykel, E.S. & Rubinsztein, D.C. (1999) Mitochondrial
genetic analyses suggest selection against maternal lineages in
bipolar affective disorder. Am J Hum Genet 65, 508518.
Krishnan, K.R. (1999) Brain imaging correlates. J Clin Psychiatry 60,
5054.

703

Yamasue et al.
Kubota, M., Kasahara, T., Nakamura, T., Ishiwata, M., Miyauchi, T. & Kato,
T. (2006) Abnormal Ca2 dynamics in transgenic mice with neuronspecific mitochondrial DNA defects. J Neurosci 26, 1231412324.
Li, Z., Okamoto, K., Hayashi, Y. & Sheng, M. (2004) The importance of
dendritic mitochondria in the morphogenesis and plasticity of
spines and synapses. Cell 119, 873887.
McMahon, F.J., Chen, Y.S., Patel, S., Kokoszka, J., Brown, M.D.,
Torroni, A., DePaulo, J.R. & Wallace, D.C. (2000) Mitochondrial
DNA sequence diversity in bipolar affective disorder. Am J Psychiatry 157, 10581064.
Minzenberg, M.J., Fan, J., New, A.S., Tang, C.Y. & Siever, L.J. (2008)
Frontolimbic structural changes in borderline personality disorder.
J Psychiatr Res 42, 727733.
Mitra, R., Jadhav, S., McEwen, B.S., Vyas, A. & Chattarji, S. (2005) Stress
duration modulates the spatiotemporal patterns of spine formation in
the basolateral amygdala. Proc Natl Acad Sci U S A 102, 93719376.
Niemi, A.K., Moilanen, J.S., Tanaka, M., Hervonen, A., Hurme, M.,
Lehtimaki, T., Arai, Y., Hirose, N. & Majamaa, K. (2005) A
combination of three common inherited mitochondrial DNA polymorphisms promotes longevity in Finnish and Japanese subjects.
Eur J Hum Genet 13, 166170.
Oldfield, R.C. (1971) The assessment and analysis of handedness: the
Edinburgh inventory. Neuropsychologia 9, 97113.
Omura, K., Todd Constable, R. & Canli, T. (2005) Amygdala gray
matter concentration is associated with extraversion and neuroticism. Neuroreport 16, 19051908.
Radley, J.J., Rocher, A.B., Miller, M., Janssen, W.G., Liston, C., Hof,
P.R., McEwen, B.S. & Morrison, J.H. (2006) Repeated stress
induces dendritic spine loss in the rat medial prefrontal cortex.
Cereb Cortex 16, 313320.
Raule, N., Sevini, F., Santoro, A., Altilia, S. & Franceschi, C. (2007)
Association studies on human mitochondrial DNA: methodological
aspects and results in the most common age-related diseases.
Mitochondrion 7, 2938.
Rizzuto, R. (2001) Intracellular Ca(2) pools in neuronal signalling.
Curr Opin Neurobiol 11, 306311.
Rizzuto, R. (2003) Calcium mobilization from mitochondria in synaptic
transmitter release. J Cell Biol 163, 441443.
Schumann, C.M., Hamstra, J., Goodlin-Jones, B.L., Lotspeich, L.J.,
Kwon, H., Buonocore, M.H., Lammers, C.R., Reiss, A.L. & Amaral,
D.G. (2004) The amygdala is enlarged in children but not adolescents with autism; the hippocampus is enlarged at all ages. J
Neurosci 24, 63926401.
Skuder, P., Plomin, R., McClearn, G.E., Smith, D.L., Vignetti, S.,
Chorney, M.J., Chorney, K., Kasarda, S., Thompson, L.A., Detterman,
D.K., Petrill, S.A., Daniels, J., Owen, M.J. & Mcguffin, P. (1995) A
polymorphism in mitochondrial DNA associated with IQ? Intelligence
21, 111.
Sterzer, P., Stadler, C., Poustka, F. & Kleinschmidt, A. (2007) A
structural neural deficit in adolescents with conduct disorder and its
association with lack of empathy. Neuroimage 37, 335342.
Strakowski, S.M., DelBello, M.P., Sax, K.W., Zimmerman, M.E.,
Shear, P.K., Hawkins, J.M. & Larson, E.R. (1999) Brain magnetic
resonance imaging of structural abnormalities in bipolar disorder.
Arch Gen Psychiatry 56, 254260.
Strakowski, S.M., Adler, C.M. & DelBello, M.P. (2002) Volumetric
MRI studies of mood disorders: do they distinguish unipolar and
bipolar disorder? Bipolar Disord 4, 8088.
Sudoyo, H., Suryadi, H., Lertrit, P., Pramoonjago, P., Lyrawati, D. &
Marzuki, S. (2002) Asian-specific mtDNA backgrounds associated
with the primary G11778A mutation of Lebers hereditary optic
neuropathy. J Hum Genet 47, 594604.

704

Sullivan, P.F. (2007) Spurious genetic associations. Biol Psychiatry 61,

11211126.
Tanaka, M., Gong, J., Zhang, J., Yamada, Y., Borgeld, H.J. & Yagi, K.
(2000) Mitochondrial genotype associated with longevity and its
inhibitory effect on mutagenesis. Mech Ageing Dev 116, 6576.
Tsuchiya, K., Miyazaki, H., Akabane, H., Yamamoto, M., Kondo, H.,
Mizusawa, H. & Ikeda, K. (1999) MELAS with prominent white
matter gliosis and atrophy of the cerebellar granular layer: a clinical,
genetic, and pathological study. Acta Neuropathol 97, 520524.
Van Erp, T.G., Saleh, P.A., Rosso, I.M., Huttunen, M., Lonnqvist, J.,
Pirkola, T., Salonen, O., Valanne, L., Poutanen, V.P., StandertskjoldNordenstam, C.G. & Cannon, T.D. (2002) Contributions of genetic
risk and fetal hypoxia to hippocampal volume in patients with
schizophrenia or schizoaffective disorder, their unaffected siblings, and healthy unrelated volunteers. Am J Psychiatry 159,
15141520.
Vermetten, E., Schmahl, C., Lindner, S., Loewenstein, R.J. & Bremner,
J.D. Hippocampal and amygdalar volumes in dissociative identity
disorder. Am J Psychiatry 163, 630636.
Vyas, A., Mitra, R., Shankaranarayana Rao, B.S. & Chattarji, S. (2002)
Chronic stress induces contrasting patterns of dendritic remodeling
in hippocampal and amygdaloid neurons. J Neurosci 22,
68106818.
Yamasue, H., Kasai, K., Iwanami, A., Ohtani, T., Yamada, H., Abe, O.,
Kuroki, N., Fukuda, R., Tochigi, M., Furukawa, S., Sadamatsu, M.,
Sasaki, T., Aoki, S., Ohtomo, K., Asukai, N. & Kato, N. (2003)
Voxel-based analysis of MRI reveals anterior cingulate gray-matter
volume reduction in posttraumatic stress disorder due to terrorism.
Proc Natl Acad Sci U S A 100, 90399043.
Yamasue, H., Iwanami, A., Hirayasu, Y., Yamada, H., Abe, O., Kuroki,
N., Fukuda, R., Tsujii, K., Aoki, S., Ohtomo, K., Kato, N. & Kasai, K.
(2004) Localized volume reduction in prefrontal, temporolimbic, and
paralimbic regions in schizophrenia: an MRI parcellation study.
Psychiatry Res 131, 195207.
Yamasue, H., Abe, O., Kasai, K., Suga, M., Iwanami, A., Yamada, H.,
Tochigi, M., Ohtani, T., Rogers, M.A., Sasaki, T., Aoki, S., Kato, T. &
Kato, N. (2007a) Human brain structural change related to acute
single exposure to sarin. Ann Neurol 61, 3746.
Yamasue, H., Abe, O., Suga, M., Yamada, H., Inoue, H., Tochigi, M.,
Rogers, M., Aoki, S., Kato, N. & Kasai, K. (2007b) Gender-common
and -specific neuroanatomical basis of human anxiety-related
personality traits. Cereb Cortex 18, 4652.
van der Walt, J.M., Nicodemus, K.K., Martin, E.R. et al. (2003)
Mitochondrial polymorphisms significantly reduce the risk of Parkinson disease. Am J Hum Genet 72, 804811.
van der Walt, J.M., Dementieva, Y.A., Martin, E.R., Scott, W.K.,
Nicodemus, K.K., Kroner, C.C., Welsh-Bohmer, K.A., Saunders,
A.M., Roses, A.D., Small, G.W., Schmechel, D.E., Murali Doraiswamy,
P., Gilbert, J.R., Haines, J.L., Vance, J.M. & Pericak-Vance, M.A.
(2004) Analysis of European mitochondrial haplogroups with
Alzheimer disease risk. Neurosci Lett 365, 2832.

Acknowledgments
This study was supported in part by grants-in-aid for scientific
research (No. 18019009 to K.K. and No. 17-5234 to M.A.R.) from
Japan Society for the Promotion of Science and the Ministry of
Education, Culture, Sports, Science and Technology, Japan, and
by grants-in-aid (H17-Kokoro-Ippan 009 to K.K.) from the Ministry
of Health, Labor and Welfare, Japan.

Genes, Brain and Behavior (2008) 7: 698704