Beruflich Dokumente
Kultur Dokumente
698
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.
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).
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.
700
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).
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
*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.
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).
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
702
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
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
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.