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

system (CNS) injuries (Arfanakis et al., 2002; De Stefano et
The underlying mechanisms leading to neurological dys- al., 2002; Parkinson et al., 2002). Unfortunately, conven-
function in white matter disorders may consist of myelin tional MRI techniques, such as T1- and T2-related measure-
dysfunction/demyelination, axonal dysfunction/injury, or ments, are not capable of differentiating axonal vs myelin
both (Li et al., 1999; McGavern et al., 1999; Perry and injury in CNS white matter disorders (Song et al., 2002).
Anthony, 1999; Trapp et al., 1999; van der Valk and De Diffusion tensor magnetic resonance imaging (DTI) has
Groot, 2000; Bjartmar et al., 2001; De Stefano et al., 2001; been widely applied in CNS for detailed developmental and
Phillips, 2001). However, the pathological processes under- pathological analyses (Filippi et al., 2001; Ito et al., 2001;
lying neurological dysfunction in white matter diseases re- Maldjian and Grossman, 2001; Nusbaum et al., 2001). After
main unclear. Magnetic resonance imaging (MRI) is a well- 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
* Corresponding author. Biomedical MR Laboratory, Campus Box
matrix diagonalization (Pierpaoli and Basser, 1996; Basser
8227, Washington University School of Medicine, 660 S. Euclid Avenue,
St. Louis, MO 63110. Fax: 1-314-362-0526. and Pierpaoli, 1998). These are scalar indices that describe
E-mail address: [email protected] (S.-K. Song). 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 and the fractional anisotropy (FA, anisotropy index normal-
tracts. ized to the magnitude of diffusion tensor). These secondary
Parameters derived from the three eigenvalues include parameters are reference frame independent and prove to be
the trace of the diffusion tensor, Tr(D) 1 2 3 sensitive to pathology (Horsfield et al., 1998; Werring et al.,
3 ADC (apparent diffusion coefficient), the relative an- 1999; Filippi et al., 2001). As secondary parameters that
isotropy (RA, the anisotropy index, defined as the standard allow a simplified expression of water diffusion, neither RA
deviation of the three eigenvalues, normalized by the ADC), (or FA) nor Tr(D) is suited for a pathology-specific analysis.
1716 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722

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
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 degeneration in optic nerve. Subsequent to this retinal de-
filled symbols represent data from the injured nerve. (A) RA, (B) Trace, generation, secondary myelin fragmentation occurs (Wal-
(C) , and (D) . Trace, , and are in units of m2/ms. RA has no lerian degeneration) (Adachi et al., 1996). A longitudinal
units. Error bars are standard deviations. 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
The current study proposes a more analytical approach to
and 7 days after the injury.
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- Materials and methods
nents parallel (1) and perpendicular (2 and 3) to the
axonal tract (Basser, 1995; Basser et al., 2000; Xue et al., Mouse model of retinal ischemia
1999). The axial diffusivity, 1 2, 3, is defined as
the magnitude of water diffusion parallel to the tract within Male Swiss Webster mice, 6 8 weeks of age (1520
the voxel of interest. Similarly, the radial diffusivity, g) were used. All procedures were in compliance with the
(2 3)/2, describes the mean ADC magnitude of water National Institute of Health Guide for the Care and Use
molecules diffusing perpendicular to the tract (Song et al., of Laboratory Animals and the ARVO Statement on the
2002). Use of Animals in Ophthalmic Research. Under anesthe-
We have hypothesized that the underlying pathology sia (a mixture of ketamine HCl, 80 mg/kg, Fort Dodge
might be discerned by evaluating directional diffusivities. Animal Health, Fort Dodge, IA, and xylazine HCl, 12
We previously showed that in dysmyelinated shiverer mg/kg, Butler Co., Columbus, OH), unilateral retinal
mouse CNS white matter, where there is little or no myelin ischemia was introduced in mice. The intraocular pres-
sheath surrounding intact axons, was not affected (Song sure (IOP) of the left eye of each mouse was raised above
et al., 2002). In contrast, the magnitude of increased, systolic blood pressure by cannulation of the anterior
reflecting the increased freedom of motion perpendicular to chamber with a 32-gauge needle connected to a saline

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.
 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722 1717

reservoir placed above the eye to produce a pressure of A conventional multislice spin-echo imaging sequence,
100 120 mm Hg (Kawai et al., 2001). The elevated IOP modified by adding the Stejskal-Tanner diffusion-sensitiz-
was maintained for 65 min (Adachi et al., 1996). Isch- ing gradient pair, was employed for acquisition of the re-
emia was confirmed by ophthalmoscopic observation of quired series of diffusion-weighted images (DWI). The dif-
the blanched fundus. The contralateral eye, which served fusion-weighted images were acquired with repetition
as the control, was not cannulated. Five male Swiss period (TR) 0.75 s, spin-echo time (TE) 50 ms, time be-
Webster mice underwent IOP elevation and longitudinal tween application of gradient pulses () 25 ms, and diffu-
DTI examination at Days 1, 3, 5, and 7 after ischemia. sion gradient duration () 10 ms. The slice thickness was 0.5
Separate groups of mice were selected at Days 3 and 7 mm, field of view (FOV) was 1.5 1.5 cm2, and data
after ischemia for cross-sectional DTI and matched his- matrix was 128 128 (zero-filled to 256 256). Diffusion-
sensitizing gradients were applied along six directions: [Gx-
tology. Three mice at each time point were examined in
,Gy,Gz] [1,1,0], [1,0,1], [0,1,1], [1,1,0], [0,1,1], and
this group.
[1,0,1]. Two diffusion-sensitizing factors, b values 0
and 0.785 ms/m2, were used for the acquisition of the
Tissue preparation DWI series (Basser and Pierpaoli, 1998). Five coronal slices
of mouse brain were collected between 0.5 to 2 mm of
Animals were killed with intraperitoneal injection of bregma.
pentobarbital (Euthasol; Delmarva Laboratories, Inc.,
Midlothian, VA) after cross-sectional DTI examinations DTI data analysis
at Days 3 and 7 after ischemia. Immediately after eutha-
nasia, both eyes were enucleated and the anterior seg- The six independent elements of the diffusion tensor
ments were removed. The posterior segments were fixed were calculated from each diffusion-weighted image. The
by immersion in 4% paraformaldehyde in 0.1 M phos- resulting tensor element maps were used to derive the eig-
phate buffer (PB, pH 7.4) for 30 min. The retinas were envalues and eigenvectors of the diffusion tensor by matrix
dissected from the choroids and treated with the same diagonalization. The quantitative indices, including Tr(D),
RA, , and were derived using software written in
fixative for 2 h at 4C. After several washes in PB, small
Matlab (MathWorks, Natick, MA). Regions of interest
pieces were cut out from the retina and then transferred to
(ROI), the cross section of optic nerves, were selected based
70% ethanol overnight. The optic nerves were dissected
on RA, , and T2-weighted (T2W) images.
and prepared as described above. The tissues were dehy-
drated with graded ethanol and then embedded in wax. Histological analyses

DTI of mouse optic nerve Myelin basic protein immunostaining was performed on
7-m-thick, rehydrated, transverse wax sections of optic
Mice were anesthetized with isoflurane/O2 (5% for in- nerves. Sections were incubated with solutions in the fol-
duction and 1.5% for maintenance). Core temperature was lowing order: 10% normal goat serum (Vector, Burlingame,
maintained at 36.5C using circulating warm water. After CA) in 0.01 M phosphate-buffered saline (PBS, pH 7.4) for
the appropriate anesthesia plane was reached, judging by the 1 h at 22C; rabbit polyclonal antibody against MBP
loss of response to tail pinch, mice were placed in a custom- (Zymed Laboratories, South San Francisco, CA) diluted
made magnetic resonance-compatible stereotaxic device to 1:100 in PBS for 16 h at 4C; goat anti-rabbit IgG conju-
immobilize the head. A 1.5-cm outer diameter, circular gated with Oregon Green (Molecular Probes, Eugene, OR)
diluted 1:800 in PBS for 30 min at 22C.
surface coil was placed on top of the head to serve as the
Similarly prepared sections were used for neurofilament
receiver for the MR signal. The combined apparatus was
immunostaining (SMI-31). Sections were incubated with
placed in a custom-made cradle permitting the brain of each
10% normal donkey serum (Vector) in PBS for 1 h at 22C;
mouse to be placed at the center of the magnet inside a 9-cm
mouse monoclonal antibody against SMI-31 (Sternberger
inner diameter Helmholtz coil (the radio frequency transmit Monoclonals, Lutherville, MD) diluted 1:2000 in PBS for
coil). 16 h at 4C; and finally with donkey anti-mouse IgG con-
The above described arrangement was positioned in a jugated with rhodamine (TRITC) (Jackson Laboratories,
second cradle that fits into an Oxford Instruments 200/330 West Grove, PA) diluted 1:200 in PBS for 30 min at 22C.
(4.7 T, 33-cm clear bore) magnet equipped with a 15-cm To detect structural change, both retina and optic nerve
inner diameter actively shielded Oxford gradient coil (20 sections were stained with hematoxylin and eosin (H&E).
G/cm, 200 s rise time). The magnet, gradient coil, and Representative sections of all samples were processed and
Oxford gradient power supply are interfaced with a Varian stained simultaneously for consistency. Control sections
UNITY-INOVA console controlled by a Sun Microsystems were prepared by eliminating primary antibody from the
Ultra-60 Sparc workstation. 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- DTI of mouse optic nerve
struments, Sterling Heights, MI) and images were stored in
Photoshop files (Adobe Photoshop; Adobe, San Jose, CA). The optic nerve is readily visible in the typical RA map
of a normal mouse brain (Fig. 1). Serial DTI measurements
Statistical analyses obtained from the five mice at 1, 3, 5, and 7 days after
reperfusion are presented in Fig. 2 with statistical compar-
Diffusion tensor parameters obtained from both eyes isons of ischemic and control eyes summarized in Table 1.
were compared using a t test. All results are expressed as There were no discernible differences in all DTI parameters
mean standard deviation (SD). The longitudinal effect of between the control and the injured optic nerves on Day 1
ischemia on DTI parameters was analyzed using analysis of after ischemia. Axial diffusivity (), RA, and Tr(D) all
variance (ANOVA). Difference was considered statistically decreased significantly in the injured optic nerve by Day 3
significant at P 0.05. 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 nation on or . Herein, we demonstrated that the optic
7 (Fig. 3B). Optic nerve damage was also observed with nerve indeed undergoes the predicted distinct patterns (Ada-
H&E staining at Days 3 and 7 after ischemia (Fig. 4). chi et al., 1996) of axonal degeneration followed by myelin
Noticeable atrophy of the injured optic nerve was observed degeneration (Figs. 2, 5, and 6), after transient retinal isch-
at Day 7 after ischemia. Visible diminution in neurofilament emia. Thus, the presentation of RA, Tr(D), , and under
immunostaining at Days 3 and 7 was apparent (Fig. 5), the influence of axonal degeneration alone or with com-
confirming the presence of axonal loss. (Recall that changes bined axonal and myelin injury can be examined using this
in also began at Day 3 after ischemia.) No discernible model.
change in MBP immunostaining at Day 3 after ischemia Effects of retinal ischemia on the optic nerve are re-
(Fig. 6) confirmed that myelin remained intact at that time, flected as early and prolonged decrease in RA. The temporal
as predicted by no change in at this time. However, evolution of Tr(D) after retinal ischemia is similar to ADC
decreased MBP intensity was apparent at Day 7 after isch- changes reported in stroke. A sharp decrease in Tr(D) is
emia as predicted by the increase in at 7 days after seen early after ischemia followed by a gradual increase
ischemia. 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
Discussion degeneration were observed (Fig. 2). These findings are
consistent with the expected and histologically documented
Both RA (or FA) and Tr(D) (or ADC) are widely used as evolution of injury to the optic nerveaxonal degeneration
summary parameters in assessing neurodegenerative dis- first, followed later by demyelination. While the integrity of
eases using DTI. The sensitivities of these parameters are the axon and the myelin sheath are not the sole determinants
well documented in both clinical and research settings of diffusion anisotropy, our findings offer strong evidence
(Horsfield et al., 1998; Werring et al., 1999; Filippi et al., that quantifying changes in and may provide a means
2001). However, these parameters do not permit specific to distinguish and monitor axonal and/or myelin pathology.
assessment of the underlying mechanisms of white matter The biphasic changes of ADC in rodent brains of stroke
injury. For example, the decrease in RA and elevation of have been reported (Fiebach et al., 2002; Helenius et al.,
ADC observed in the normal appearing white matter from 2002; Sotak, 2002). The initial decrease in ADC may be
multiple sclerosis (MS) patients do not distinguish axonal caused by cytotoxic edema (Fiebach et al., 2002) and other
injury vs demyelination (Horsfield et al., 1998; Werring et cellular changes. The elevation of ADC at a later time to its
al., 1999; van der Valk and De Groot, 2000; Filippi et al., normal or supranormal value is attributed to tissue loss
2001; Maldjian and Grossman, 2001). following disintegration caused by the ischemic insult.
Although CNS demyelination is considered the patho- However, this line of interpretation may not apply to the
logic hallmark in MS, historical and recent studies have injured optic nerve from mice following retinal ischemia.
documented early pathological changes in axons. Loss of Our results show the combined changes of and con-
myelin in CNS white matter may not interrupt conduction tribute to the observed Tr(D) (or ADC) evolution in the
and has the potential for repair (remyelination). Accumu- white matter. The normalization of Tr(D) beginning after 5
lating evidence suggests that axon loss may be a principal days of reperfusion is the consequence of elevated off-
reason for irreversible neurological disability in MS patients setting the decrease of (Fig. 2). Note that in this cohort
(Trapp et al., 1999; van der Valk and De Groot, 2000; De of animals does show a trend of normalization at Day 7, but
Stefano et al., 2001, 2002; Phillips, 2001). If proven in the this was not statistically different compared to at Day 5.
clinical setting, a noninvasive means to differentiate these Thus, the present data indicate that Tr(D) (or ADC) evolu-
pathologies such as we describe here would present a valu- tion cannot be generalized to both gray and white matter in
able therapeutic opportunity to target the specific origin of CNS injury.
the neurological dysfunction. Recently, Piepaoli et al. reported the use of DTI to
In a recent study, we demonstrated that was signifi- differentiate primary injury and Wallerian degeneration
cantly higher in shiverer mouse brain white matter com- (Pierpaoli et al., 2001) of CNS white matter. They found
pared with age-matched controls, reflecting the lack of my- that secondary white matter degeneration was revealed by a
elin and the increased freedom of cross-fiber diffusion in the large reduction in diffusion anisotropy only in regions
dysmyelinated white matter (Song et al., 2002). was not where fibers are arranged in isolated bundles of parallel
different between shiverer and control mice, consistent with fibers, such as in the cerebral peduncle. In regions where the
the presence of intact axons. Based on these previous find- degenerated pathway crosses other tracts, there was almost
ings, we hypothesize that changes in and may poten- no change in diffusion anisotropy, but a significant change
tially be used to differentiate myelin loss vs axonal injury. in the measured orientation of fibers. Although still to be
To further test our hypothesis, a mouse model of tran- proved, we believe that their ability to detect the significant
sient retinal ischemia was used in the present studies to change in the measured orientation of fibers crossing other
determine the effects of axonal degeneration and demyeli- tracts may be an indication that directional diffusivity may
 S.-K. Song et al. / NeuroImage 20 (2003) 1714 1722 1721

still be able to differentiate axonal vs myelin degeneration. Basser, P.J., 1995. Inferring microstructural features and the physiological
In our own studies, we have detected changes of directional state of tissues from diffusion-weighted images. NMR Biomed. 8,
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Acknowledgments Kim, G.M., Xu, J., Song, S.K., Yan, P., Ku, G., Xu, X.M., Hsu, C.Y., 2001.
Tumor necrosis factor receptor deletion reduces nuclear factor-kappaB
activation. cellular inhibitor of apoptosis protein 2 expression, and
This study was supported in part by the National Multi- functional recovery after traumatic spinal cord injury. J. Neurosci. 21,
ple Sclerosis Society (RG 3376-A-2/1, and CA 1012-A-13), 6617 6625.
the Washington University Small Animal Imaging Resource Li, S., Mealing, G.A., Morley, P., Stys, P.K., 1999. Novel injury mecha-
(WUSAIR), a National Cancer Institute supported Small nism in anoxia and trauma of spinal cord white matter: glutamate
Animal Imaging Resource Program (SAIRP) center (NIH release via reverse Na-dependent glutamate transport. J. Neurosci. 19,
RC16.
Grant R24-CA83060), and NIH grant EY12017 (A.H.N.).
Maldjian, J.A., Grossman, R.I., 2001. Future applications of DWI in MS.
Helpful discussions with Drs. Jeff J. Neil, Joseph J. H. J. Neurol. Sci. 186 (Suppl 1), S55S57.
Ackerman, and other members of the Washington Univer- McGavern, D.B., Murray, P.D., Rodriguez, M., 1999. Quantitation of
sity Biomedical MR Laboratory are gratefully acknowl- spinal cord demyelination, remyelination, atrophy, and axonal loss in a
edged. model of progressive neurologic injury. J. Neurosci. Res. 58, 492504.
Nusbaum, A.O., Tang, C.Y., Buchsbaum, M.S., Wei, T.C., Atlas, S.W.,
2001. Regional and global changes in cerebral diffusion with normal
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