NeuroImage 20 (2003) 1714 1722 www.elsevier.

com/locate/ynimg

Diffusion tensor imaging detects and differentiates axon and myelin

degeneration in mouse optic nerve after retinal ischemia
Sheng-Kwei Song,a,* Shu-Wei Sun,a Won-Kyu Ju,b Shiow-Jiuan Lin,c
Anne H. Cross,d and Arthur H. Neufeldb
a
Department of Radiology, Washington University School of Medicine, St. Louis, MO 63110, USA,
b
Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St. Louis, MO 63110, USA,
c
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA
d
Department of Neurology and Neurosurgery, Washington University School of Medicine, St. Louis, MO 63110, USA

Received 8 April 2003; revised 8 July 2003; accepted 14 July 2003

Abstract
Both axon and myelin degeneration have significant impact on the long-term disability of patients with white matter disorder. However,
the clinical manifestations of the neurological dysfunction caused by white matter disorders are not sufficient to determine the origin of
neurological deficits. A noninvasive biological marker capable of detecting and differentiating axon and myelin degeneration would be a
significant addition to currently available tools. Directional diffusivities derived from diffusion tensor imaging (DTI) have been previously
proposed by this group as potential biological markers to detect and differentiate axon and myelin degeneration. To further test the
hypothesis that axial () and radial () diffusivities reflect axon and myelin pathologies, respectively, the optic nerve was examined
serially using DTI in a mouse model of retinal ischemia. A significant decrease of , the putative DTI axonal marker, was observed 3 days
after ischemia without concurrently detectable changes in , the putative myelin marker. This result is consistent with histological findings
of significant axonal degeneration with no detectable demyelination at 3 days after ischemia. The elevation of observed 5 days after
ischemia is consistent with histological findings of myelin degeneration at this time. These results support the hypothesis that and hold
promise as specific markers of axonal and myelin injury, respectively, and, further, that the coexistence of axonal and myelin degeneration
does not confound this utility.
2003 Elsevier Inc. All rights reserved.

Introduction established method for detecting various central nervous

The underlying mechanisms leading to neurological dys-
function in white matter disorders may consist of myelin
dysfunction/demyelination, axonal dysfunction/injury, or
both (Li et al., 1999; McGavern et al., 1999; Perry and
Anthony, 1999; Trapp et al., 1999; van der Valk and De
Groot, 2000; Bjartmar et al., 2001; De Stefano et al., 2001;
Phillips, 2001). However, the pathological processes under-
lying neurological dysfunction in white matter diseases re-
main unclear. Magnetic resonance imaging (MRI) is a well-
* Corresponding author. Biomedical MR Laboratory, Campus Box
8227, Washington University School of Medicine, 660 S. Euclid Avenue,
St. Louis, MO 63110. Fax: 1-314-362-0526.
E-mail address: victor@wuchem.wustl.edu (S.-K. Song).
system (CNS) injuries (Arfanakis et al., 2002; De Stefano et
al., 2002; Parkinson et al., 2002). Unfortunately, conven-
tional MRI techniques, such as T1- and T2-related measure-
ments, are not capable of differentiating axonal vs myelin
injury in CNS white matter disorders (Song et al., 2002).
Diffusion tensor magnetic resonance imaging (DTI) has
been widely applied in CNS for detailed developmental and
pathological analyses (Filippi et al., 2001; Ito et al., 2001;
Maldjian and Grossman, 2001; Nusbaum et al., 2001). After
generation of the diffusion tensor matrix from a series of
diffusion-weighted images, the three eigenvalues, also re-
ferred to as diffusivities (1, 2, and 3), are calculated by
matrix diagonalization (Pierpaoli and Basser, 1996; Basser
and Pierpaoli, 1998). These are scalar indices that describe
water diffusion in a local, i.e., voxel specific, frame of

1053-8119/$ see front matter 2003 Elsevier Inc. All rights reserved.
doi:10.1016/j.neuroimage.2003.07.005
 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722 1715

Fig. 1. Relative anisotropy map of a coronal slice of mouse brain contains optic nerves (red square). The region of optic nerve is expanded to show that the
proposed measurement on mouse optic nerve is feasible (i.e., there is a sufficient voxel density within the imaged cross section of each optic nerve).
Fig. 4. H&E staining of the mouse optic nerve. Panels A and B are from the optic nerve of a contralateral eye at Day 3 after ischemia. Panels C and D are
from the optic nerve of the ipsilateral eye at Day 3 after ischemia. High magnification shows severe axonal degeneration. Panels E and F are optic nerves
at Day 7 after ischemia. (A, C and E) Scale bar, 100 m; (B, D, and F) scale bar, 20 m.

reference that coincides with the geometry of white matter
tracts.
Parameters derived from the three eigenvalues include
the trace of the diffusion tensor, Tr(D) 1 2 3
3 ADC (apparent diffusion coefficient), the relative an-
isotropy (RA, the anisotropy index, defined as the standard
deviation of the three eigenvalues, normalized by the ADC),
and the fractional anisotropy (FA, anisotropy index normal-
ized to the magnitude of diffusion tensor). These secondary
parameters are reference frame independent and prove to be
sensitive to pathology (Horsfield et al., 1998; Werring et al.,
1999; Filippi et al., 2001). As secondary parameters that
allow a simplified expression of water diffusion, neither RA
(or FA) nor Tr(D) is suited for a pathology-specific analysis.
1716 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722
Fig. 2. Time course of averaged DTI parameters of optic nerves from five
mice. The open symbols are the data from the control optic nerve and the
filled symbols represent data from the injured nerve. (A) RA, (B) Trace,
(C) , and (D) . Trace, , and are in units of m2/ms. RA has no
units. Error bars are standard deviations.
The current study proposes a more analytical approach to
obtain specific information defining the underlying pathol-
ogy in white matter injury.
The eigenvalues (1, 2, and 3) derived by diffusion
tensor matrix diagonalization can be separated into compo-
nents parallel (1) and perpendicular (2 and 3) to the
axonal tract (Basser, 1995; Basser et al., 2000; Xue et al.,
1999). The axial diffusivity, 1 2, 3, is defined as
the magnitude of water diffusion parallel to the tract within
the voxel of interest. Similarly, the radial diffusivity,
(2 3)/2, describes the mean ADC magnitude of water
molecules diffusing perpendicular to the tract (Song et al.,
2002).
We have hypothesized that the underlying pathology
might be discerned by evaluating directional diffusivities.
We previously showed that in dysmyelinated shiverer
mouse CNS white matter, where there is little or no myelin
sheath surrounding intact axons, was not affected (Song
et al., 2002). In contrast, the magnitude of increased,
reflecting the increased freedom of motion perpendicular to
Fig. 3. H&E staining of the normal retina (A) and the retina on Day 7 after ischemia (B). There is marked loss of the IPL and GCL in the retina after ischemia.
Abbreviations: ONL, outer nuclear layer; OPL, outer plexiform layer; INL, inner nuclear layer; IPL, inner plexiform layer; and GCL, ganglion cell layer.
Scale bar, 20 m.
axons due to the lack of myelin (Song et al., 2002). It is also
our hypothesis that axonal degeneration without (or prior to)
demyelination will result in a decrease in both and .
The degree of decrease in is expected to be relatively
much smaller than that in and is less likely to be detect-
able experimentally since is 10 30% of in magni-
tude. Thus, we propose that axonal degeneration in the
absence of demyelination will result in a detectable decrease
of .
To test our hypothesis, we used a mouse model of retinal
ischemia that causes an acute inner retinal degeneration
(Kawai et al., 2001; Rosenbaum et al., 2001) with axonal
degeneration in optic nerve. Subsequent to this retinal de-
generation, secondary myelin fragmentation occurs (Wal-
lerian degeneration) (Adachi et al., 1996). A longitudinal
DTI examination of the optic nerve in this mouse model was
conducted and correlated with neurofilament and myelin
basic protein (MBP) immunostaining (Kim et al., 2001) at 3
and 7 days after the injury.
Materials and methods
Mouse model of retinal ischemia
Male Swiss Webster mice, 6 8 weeks of age (1520
g) were used. All procedures were in compliance with the
National Institute of Health Guide for the Care and Use
of Laboratory Animals and the ARVO Statement on the
Use of Animals in Ophthalmic Research. Under anesthe-
sia (a mixture of ketamine HCl, 80 mg/kg, Fort Dodge
Animal Health, Fort Dodge, IA, and xylazine HCl, 12
mg/kg, Butler Co., Columbus, OH), unilateral retinal
ischemia was introduced in mice. The intraocular pres-
sure (IOP) of the left eye of each mouse was raised above
systolic blood pressure by cannulation of the anterior
chamber with a 32-gauge needle connected to a saline
 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722
reservoir placed above the eye to produce a pressure of
100 120 mm Hg (Kawai et al., 2001). The elevated IOP
was maintained for 65 min (Adachi et al., 1996). Isch-
emia was confirmed by ophthalmoscopic observation of
the blanched fundus. The contralateral eye, which served
as the control, was not cannulated. Five male Swiss
Webster mice underwent IOP elevation and longitudinal
DTI examination at Days 1, 3, 5, and 7 after ischemia.
Separate groups of mice were selected at Days 3 and 7
after ischemia for cross-sectional DTI and matched his-
tology. Three mice at each time point were examined in
this group.
Tissue preparation
Animals were killed with intraperitoneal injection of
pentobarbital (Euthasol; Delmarva Laboratories, Inc.,
Midlothian, VA) after cross-sectional DTI examinations
at Days 3 and 7 after ischemia. Immediately after eutha-
nasia, both eyes were enucleated and the anterior seg-
ments were removed. The posterior segments were fixed
by immersion in 4% paraformaldehyde in 0.1 M phos-
phate buffer (PB, pH 7.4) for 30 min. The retinas were
dissected from the choroids and treated with the same
fixative for 2 h at 4C. After several washes in PB, small
pieces were cut out from the retina and then transferred to
70% ethanol overnight. The optic nerves were dissected
and prepared as described above. The tissues were dehy-
drated with graded ethanol and then embedded in wax.
DTI of mouse optic nerve
Mice were anesthetized with isoflurane/O2 (5% for in-
duction and 1.5% for maintenance). Core temperature was
maintained at 36.5C using circulating warm water. After
the appropriate anesthesia plane was reached, judging by the
loss of response to tail pinch, mice were placed in a custom-
made magnetic resonance-compatible stereotaxic device to
immobilize the head. A 1.5-cm outer diameter, circular
surface coil was placed on top of the head to serve as the
receiver for the MR signal. The combined apparatus was
placed in a custom-made cradle permitting the brain of each
mouse to be placed at the center of the magnet inside a 9-cm
inner diameter Helmholtz coil (the radio frequency transmit
coil).
The above described arrangement was positioned in a
second cradle that fits into an Oxford Instruments 200/330
(4.7 T, 33-cm clear bore) magnet equipped with a 15-cm
inner diameter actively shielded Oxford gradient coil (20
G/cm, 200 s rise time). The magnet, gradient coil, and
Oxford gradient power supply are interfaced with a Varian
UNITY-INOVA console controlled by a Sun Microsystems
Ultra-60 Sparc workstation.
1717
A conventional multislice spin-echo imaging sequence,
modified by adding the Stejskal-Tanner diffusion-sensitiz-
ing gradient pair, was employed for acquisition of the re-
quired series of diffusion-weighted images (DWI). The dif-
fusion-weighted images were acquired with repetition
period (TR) 0.75 s, spin-echo time (TE) 50 ms, time be-
tween application of gradient pulses () 25 ms, and diffu-
sion gradient duration () 10 ms. The slice thickness was 0.5
mm, field of view (FOV) was 1.5 1.5 cm2, and data
matrix was 128 128 (zero-filled to 256 256). Diffusion-
sensitizing gradients were applied along six directions: [Gx-
,Gy,Gz] [1,1,0], [1,0,1], [0,1,1], [1,1,0], [0,1,1], and
[1,0,1]. Two diffusion-sensitizing factors, b values 0
and 0.785 ms/m2, were used for the acquisition of the
DWI series (Basser and Pierpaoli, 1998). Five coronal slices
of mouse brain were collected between 0.5 to 2 mm of
bregma.
DTI data analysis
The six independent elements of the diffusion tensor
were calculated from each diffusion-weighted image. The
resulting tensor element maps were used to derive the eig-
envalues and eigenvectors of the diffusion tensor by matrix
diagonalization. The quantitative indices, including Tr(D),
RA, , and were derived using software written in
Matlab (MathWorks, Natick, MA). Regions of interest
(ROI), the cross section of optic nerves, were selected based
on RA, , and T2-weighted (T2W) images.
Histological analyses
Myelin basic protein immunostaining was performed on
7-m-thick, rehydrated, transverse wax sections of optic
nerves. Sections were incubated with solutions in the fol-
lowing order: 10% normal goat serum (Vector, Burlingame,
CA) in 0.01 M phosphate-buffered saline (PBS, pH 7.4) for
1 h at 22C; rabbit polyclonal antibody against MBP
(Zymed Laboratories, South San Francisco, CA) diluted
1:100 in PBS for 16 h at 4C; goat anti-rabbit IgG conju-
gated with Oregon Green (Molecular Probes, Eugene, OR)
diluted 1:800 in PBS for 30 min at 22C.
Similarly prepared sections were used for neurofilament
immunostaining (SMI-31). Sections were incubated with
10% normal donkey serum (Vector) in PBS for 1 h at 22C;
mouse monoclonal antibody against SMI-31 (Sternberger
Monoclonals, Lutherville, MD) diluted 1:2000 in PBS for
16 h at 4C; and finally with donkey anti-mouse IgG con-
jugated with rhodamine (TRITC) (Jackson Laboratories,
West Grove, PA) diluted 1:200 in PBS for 30 min at 22C.
To detect structural change, both retina and optic nerve
sections were stained with hematoxylin and eosin (H&E).
Representative sections of all samples were processed and
stained simultaneously for consistency. Control sections
were prepared by eliminating primary antibody from the
incubation medium. Slides were examined using a micro-
1718 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722

Fig. 5. Neurofilament (SMI-31) labeling of mouse optic nerve. Panels A and B are from the contralateral optic nerve of a control eye at Day 3 after ischemia.
Panels C and D are from the ipsilateral optic nerve of the ischemic eye at Day 3 after ischemia. Panels E and F are from the ipsilateral optic nerve on Day
7 after ischemia. High magnification shows changes in labeling intensities of the optic nerves on Days 3 and 7 after ischemia. (A, C, and E). Scale bar, 100
m; (B, D, and F) scale bar, 20 m.

scope (AX70; Olympus, Tokyo, Japan). Fluorescence and Results

differential interference contrast (DIC) images were re-
corded by digital photomicrography (Spot; Diagnostic In-
struments, Sterling Heights, MI) and images were stored in
Photoshop files (Adobe Photoshop; Adobe, San Jose, CA).
Statistical analyses
Diffusion tensor parameters obtained from both eyes
were compared using a t test. All results are expressed as
mean standard deviation (SD). The longitudinal effect of
ischemia on DTI parameters was analyzed using analysis of
variance (ANOVA). Difference was considered statistically
significant at P 0.05.
DTI of mouse optic nerve
The optic nerve is readily visible in the typical RA map
of a normal mouse brain (Fig. 1). Serial DTI measurements
obtained from the five mice at 1, 3, 5, and 7 days after
reperfusion are presented in Fig. 2 with statistical compar-
isons of ischemic and control eyes summarized in Table 1.
There were no discernible differences in all DTI parameters
between the control and the injured optic nerves on Day 1
after ischemia. Axial diffusivity (), RA, and Tr(D) all
decreased significantly in the injured optic nerve by Day 3
after ischemia with the largest change seen in axial diffu-
 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722 1719

Fig. 6. MBP labeling of mouse optic nerve. Panels A and B are from the contralateral optic nerve of a control eye at Day 3 after ischemia. Panels C and
D are from the ipsilateral optic nerve of the ischemic eye at Day 3 after ischemia. Panels E and F are from the ipsilateral optic nerve on Day 7 after ischemia.
Changes in labeling intensities of the optic nerves are not apparent until Day 7 after ischemia. (A, C, and E) Scale bar, 100 m; (B, D and F) scale bar, 20
m.

sivity. There was no detectable change in between con- Day 3 and stayed at this level on Day 5 after ischemia. It
trol and injured optic nerves until Day 5 after ischemia. Fig. then reversed the course and returned to control value by
2 plots the evolution in DTI parameters in the ischemic and Day 7 after ischemia (Fig. 2B). The value of reached its
control eyes. RA reached its lowest value on Day 5 and minimum on Day 3 after ischemia and remained constant on
maintained this level on Day 7 after ischemia (Fig. 2A). Days 5 and 7 after ischemia (Fig. 2C). The upward trend
Differences in RA between Days 5 and 7 were not statisti- between Days 5 and 7 of was not statistically significant.
cally significant. Tr(D) decreased to a minimum value on There was no detectable change in until Day 5 after
ischemia. It continued to increase significantly at Day 7
after ischemia (Fig. 2D).
Table 1
Summary of paired t test P values for ischemic and control optic nerves
Histological findings
RA Tr(D)
Day 1 0.88 0.75 0.76 0.77 Optic nerves and eyes were excised at the conclusion of
Day 3 0.04 0.001 0.001 0.22 the cross-sectional in vivo DTI examinations at Days 3 and
Day 5 0.001* 0.001 0.001 0.03* 7 after ischemia (three pairs at each time point). Retinal
Day 7 0.0001 0.06 0.001 0.01*
degeneration, with severe damage of the inner plexiform
* Unequal variance. layer (IPL) and ganglion cell layer (GCL) caused by the
1720 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722
ischemic insult, was demonstrated by H&E staining on Day
7 (Fig. 3B). Optic nerve damage was also observed with
H&E staining at Days 3 and 7 after ischemia (Fig. 4).
Noticeable atrophy of the injured optic nerve was observed
at Day 7 after ischemia. Visible diminution in neurofilament
immunostaining at Days 3 and 7 was apparent (Fig. 5),
confirming the presence of axonal loss. (Recall that changes
in also began at Day 3 after ischemia.) No discernible
change in MBP immunostaining at Day 3 after ischemia
(Fig. 6) confirmed that myelin remained intact at that time,
as predicted by no change in at this time. However,
decreased MBP intensity was apparent at Day 7 after isch-
emia as predicted by the increase in at 7 days after
ischemia.
Discussion
Both RA (or FA) and Tr(D) (or ADC) are widely used as
summary parameters in assessing neurodegenerative dis-
eases using DTI. The sensitivities of these parameters are
well documented in both clinical and research settings
(Horsfield et al., 1998; Werring et al., 1999; Filippi et al.,
2001). However, these parameters do not permit specific
assessment of the underlying mechanisms of white matter
injury. For example, the decrease in RA and elevation of
ADC observed in the normal appearing white matter from
multiple sclerosis (MS) patients do not distinguish axonal
injury vs demyelination (Horsfield et al., 1998; Werring et
al., 1999; van der Valk and De Groot, 2000; Filippi et al.,
2001; Maldjian and Grossman, 2001).
Although CNS demyelination is considered the patho-
logic hallmark in MS, historical and recent studies have
documented early pathological changes in axons. Loss of
myelin in CNS white matter may not interrupt conduction
and has the potential for repair (remyelination). Accumu-
lating evidence suggests that axon loss may be a principal
reason for irreversible neurological disability in MS patients
(Trapp et al., 1999; van der Valk and De Groot, 2000; De
Stefano et al., 2001, 2002; Phillips, 2001). If proven in the
clinical setting, a noninvasive means to differentiate these
pathologies such as we describe here would present a valu-
able therapeutic opportunity to target the specific origin of
the neurological dysfunction.
In a recent study, we demonstrated that was signifi-
cantly higher in shiverer mouse brain white matter com-
pared with age-matched controls, reflecting the lack of my-
elin and the increased freedom of cross-fiber diffusion in the
dysmyelinated white matter (Song et al., 2002). was not
different between shiverer and control mice, consistent with
the presence of intact axons. Based on these previous find-
ings, we hypothesize that changes in and may poten-
tially be used to differentiate myelin loss vs axonal injury.
To further test our hypothesis, a mouse model of tran-
sient retinal ischemia was used in the present studies to
determine the effects of axonal degeneration and demyeli-
nation on or . Herein, we demonstrated that the optic
nerve indeed undergoes the predicted distinct patterns (Ada-
chi et al., 1996) of axonal degeneration followed by myelin
degeneration (Figs. 2, 5, and 6), after transient retinal isch-
emia. Thus, the presentation of RA, Tr(D), , and under
the influence of axonal degeneration alone or with com-
bined axonal and myelin injury can be examined using this
model.
Effects of retinal ischemia on the optic nerve are re-
flected as early and prolonged decrease in RA. The temporal
evolution of Tr(D) after retinal ischemia is similar to ADC
changes reported in stroke. A sharp decrease in Tr(D) is
seen early after ischemia followed by a gradual increase
toward the control level at Day 7. Distinct evolution patterns
of (early and prolonged decrease) and (delayed and
sustained elevation) during the progression of optic nerve
degeneration were observed (Fig. 2). These findings are
consistent with the expected and histologically documented
evolution of injury to the optic nerveaxonal degeneration
first, followed later by demyelination. While the integrity of
the axon and the myelin sheath are not the sole determinants
of diffusion anisotropy, our findings offer strong evidence
that quantifying changes in and may provide a means
to distinguish and monitor axonal and/or myelin pathology.
The biphasic changes of ADC in rodent brains of stroke
have been reported (Fiebach et al., 2002; Helenius et al.,
2002; Sotak, 2002). The initial decrease in ADC may be
caused by cytotoxic edema (Fiebach et al., 2002) and other
cellular changes. The elevation of ADC at a later time to its
normal or supranormal value is attributed to tissue loss
following disintegration caused by the ischemic insult.
However, this line of interpretation may not apply to the
injured optic nerve from mice following retinal ischemia.
Our results show the combined changes of and con-
tribute to the observed Tr(D) (or ADC) evolution in the
white matter. The normalization of Tr(D) beginning after 5
days of reperfusion is the consequence of elevated off-
setting the decrease of (Fig. 2). Note that in this cohort
of animals does show a trend of normalization at Day 7, but
this was not statistically different compared to at Day 5.
Thus, the present data indicate that Tr(D) (or ADC) evolu-
tion cannot be generalized to both gray and white matter in
CNS injury.
Recently, Piepaoli et al. reported the use of DTI to
differentiate primary injury and Wallerian degeneration
(Pierpaoli et al., 2001) of CNS white matter. They found
that secondary white matter degeneration was revealed by a
large reduction in diffusion anisotropy only in regions
where fibers are arranged in isolated bundles of parallel
fibers, such as in the cerebral peduncle. In regions where the
degenerated pathway crosses other tracts, there was almost
no change in diffusion anisotropy, but a significant change
in the measured orientation of fibers. Although still to be
proved, we believe that their ability to detect the significant
change in the measured orientation of fibers crossing other
tracts may be an indication that directional diffusivity may
 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722
still be able to differentiate axonal vs myelin degeneration.
In our own studies, we have detected changes of directional
diffusivities in almost all white matter tracts in the brain of
a transgenic mouse over expressing -amyloid precursor
protein under control of platelet-derived growth factor pro-
moter (unpublished results). To avoid the complication of
crossing fibers at this preliminary stage of the study, we
chose to examine the simple fiber of optic nerve using the
mouse model of retinal ischemia.
Although more comprehensive investigation is still
needed, our research into the effect of axonal degeneration
on indicates that the decrease in caused by axonal
degeneration is exceedingly small and typically below the
interanimal differences. The decrease in at Day 3 after
reperfusion, when only axonal degeneration is present his-
tologically, was about 8% of the control value in the present
study, whereas the standard deviation of between ani-
mals is in the range of 10 30% of the mean . On the
other hand, the elevation of caused by demyelination
was readily detectable, typically greater than 30% of base-
line. In the current retinal ischemia mouse model, the in
the injured optic nerve was about 40 and 100% larger than
that of the control optic nerve at Days 5 and 7, respectively,
after reperfusion. This improved dynamic range of in
detecting myelin degeneration may be feasible for use in the
diagnosis of white matter disorders in both research and
clinical settings.
In summary, we have used a mouse model of retinal
ischemia to demonstrate differential changes in axial and
radial diffusivities in a white matter tract by DTI. These
changes, taken together with histological findings, suggest
that and are markers of axonal and myelin injury,
respectively.
Acknowledgments
This study was supported in part by the National Multi-
ple Sclerosis Society (RG 3376-A-2/1, and CA 1012-A-13),
the Washington University Small Animal Imaging Resource
(WUSAIR), a National Cancer Institute supported Small
Animal Imaging Resource Program (SAIRP) center (NIH
Grant R24-CA83060), and NIH grant EY12017 (A.H.N.).
Helpful discussions with Drs. Jeff J. Neil, Joseph J. H.
Ackerman, and other members of the Washington Univer-
sity Biomedical MR Laboratory are gratefully acknowl-
edged.
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