Detection of postmortem human cerebellar cortex and white matter
pathways using high angular resolution diffusion tractography:
A feasibility study
Emi Takahashia,b,c,⁎, Jae W. Songa,b, Rebecca D. Folkerthd,e, P. Ellen Granta,b,c,f, Jeremy D. Schmahmanng
aDivision of Newborn Medicine, Department of Medicine, Boston Children's Hospital, Harvard Medical School, Boston MA, USA
bFetal-Neonatal Neuroimaging and Developmental Science Center, Boston MA, USA
cAthinoula A. Martinos Center for Biomedical Imaging, Charlestown, MA, USA
dDepartment of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston MA, USA
eDepartment of Pathology, Boston Children's Hospital, Harvard Medical School, Boston MA, USA
fDepartment of Radiology, Boston Children's Hospital, Harvard Medical School, Boston MA, USA
gAtaxia Unit, Cognitive and Behavioral Neurology Unit, Laboratory for Neuroanatomy and Cerebellar Neurobiology, Department of Neurology, Massachusetts General Hospital, Harvard
Medical School, Boston, MA, USA
a b s t r a c ta r t i c l e i n f o
Accepted 21 November 2012
Available online 11 December 2012
Imaging three-dimensional cerebellar connectivity using diffusion tractography is challenging because of the
ubiquitous features of crossing axonal pathways within a folium as well as intersecting pathways from neigh-
boring folia. We applied high-angular resolution diffusion imaging (HARDI) tractography to intact postmor-
tem adult brainstem and cerebellum to examine the 3-dimensional white matter and local gray matter
pathways. The middle cerebellar peduncles conveyed fibers from the rostral pons to the lateral and caudal
aspects of the cerebellar hemisphere, and from the caudal pons to medial and rostral parts of the cerebellar
hemisphere. In the cerebellar cortex, tractography detected tangential coherence superficially in the cerebel-
lar cortex and revealed fibers coursing parallel to the long axis of the folia. These fibers were consistent with
the location and direction of parallel fibers in the molecular layer. Crossing with these parallel fibers were
tangential fibers running perpendicular to the long axis of the folia, consistent with axons of the cortical
interneurons — stellate cells and basket cells. These tangential fibers within the cerebellar cortex were dis-
tinct from the fibers linking the cerebellar cortex with the deep cerebellar nuclei and the brainstem. Our re-
sults show the potential for HARDI tractography to resolve axonal pathways from different neuronal
elements within the cerebellar cortex, and improve our understanding of adult cerebellar neural circuitry
and connectivity in both white and gray matter.
© 2012 Elsevier Inc. All rights reserved.
The cerebellum is critical for sensorimotor function, as well as for
intellectual processing and emotional regulation (Schmahmann, 1997,
2010). Anatomical studies of the cerebellum and its connections with
the cerebral hemispheres in the non-human primate have helped de-
fine the neurobiological underpinnings of these relationships (Brodal
and Bjaalie, 1997; Schmahmann and Pandya, 1997; Strick et al., 2009).
The development of diffusion tractography enables the study of these
connections non-invasively in animal models and in the human brain.
Diffusion tensor imaging (DTI) studies of the developing cerebellum
(Huang et al., 2009; Saksena et al., 2008) have shown the location and
trajectories of the major cerebellar white matter pathways, and these
new imaging approaches hold the promise of a deeper understanding
of the anatomical basis of cerebellar functions in the human brain.
Identification of the complex 3-dimensional patterns of intracerebellar
and extracerebellar connections using diffusion tractography is limited,
however, because of the multiple crossing axonal pathways in the cer-
ebellar white matter and cortex, and because detection of tractography
paths within a folium is often contaminated by paths from the closely
apposed and narrow neighboring folia.
High-angular resolution diffusion imaging (HARDI) improves the
of its abilitytodefinefiberorientation distribution functions(Leergaard
different diffusion directions within the same voxel resulting from
crossing axonal bundles. We have previously used HARDI tractography
successfully to resolve white matter pathways in the cerebellum in vivo
(Granziera et al., 2009) and in the cerebrum ex vivo (Takahashi et al.,
2011, 2012). High-resolution tractography using HARDI has not yet
been applied to the study of postmortem human cerebellum. In this
tem adult human cerebellum to examinethe 3-dimensional fiber struc-
tures in cerebellar white matter and local gray matter.
NeuroImage 68 (2013) 105–111
⁎ Corresponding author at: Division of Newborn Medicine, Department of Medicine,
Boston Children's Hospital, Harvard Medical School, USA.
E-mail address: email@example.com (E. Takahashi).
1053-8119/$ – see front matter © 2012 Elsevier Inc. All rights reserved.
Contents lists available at SciVerse ScienceDirect
journal homepage: www.elsevier.com/locate/ynimg
The advantages of ex vivo imaging include the lack of motion that
enables imaging at very high resolution for the long periods of time
necessary to obtain adequate signal to noise for HARDI, the absence
of blood flow as a confounding factor, and the lack of susceptibility ar-
tifacts at air or bone and brain interfaces. These advantages of ex vivo
imaging are critical when trying to image fibers in the cerebellar cor-
tex because the cortex is thin, and because fibers in the cerebellar
cortex are less coherent than those in the white matter. HARDI is
well-suited for identifying pathways in the cerebellar gray matter be-
cause different types of cells are regularly aligned in the cerebellar
cortex and the topographical relationships between the cells and
folia are common across regions. In addition to the advantages of
imaging postmortem tissues and the naturally coherent and regular
cellularorganizationof thecerebellum,anoptimal MRcoil cansubstan-
tially improve signal to noiseand the abilityof diffusion tractographyto
identify gray matter structure. MR signal strength is proportional to the
distance from the MR coil. The location of the cerebellum within the
whole brain therefore presents a challenge to obtaining optimal signal
to noise and spatial resolution. In this postmortem study we dissected
the cerebellum and brainstem off from the cerebrum, and used an MR
coil closely apposed to the surface of the cerebellum, thus enabling
the highest possible signal to noise ratio and spatial resolution.
We performed MR scans on three specimens of the adult human
cerebellum. The cerebellums were acquired from the Brigham and
by the hospital's institutional review board for human research. A neu-
ropathologist studied each brain at the time of post-mortem examina-
tion, and only those not needed for immediate neuropathological
diagnosis were fixed in 4% paraformaldehyde and submitted for coded
(de-identified) specimen scanning. Mean fixation period was approxi-
mately 2–3 months. Any cases with known or suspected malformations,
disruptions, or other lesions were excluded from this study. Brains were
removed from the cranium and fixed in a 4% paraformaldehyde solution
containing 1 mM gadolinium (Gd-DTPA) MRI contrast agent for at least
1 week to reduce the T1 relaxation time while ensuring that sufficient
T2-weighted signal remains. Before image acquisition, the cerebellum
and pons were separated from the cerebral hemispheres by transverse
section through the midbrain at the level of the colliculi. During image
lution (Ausimont, Thorofare, NJ) and imaged on a 4.7 T Bruker Biospec
MR system, using an MR coil (10 cm diameter) in order to maximize
the signal to noise ratio.
The pulse sequence used for HARDI acquisition was a 3D diffusion-
weighted spin-echo echo-planar imaging (EPI) sequence, TR/TE
1000/40 ms, with an imaging matrix of 128×128×112 pixels. Sixty
diffusion-weighted measurements and one non diffusion-weighted
measurement were acquired at b=4000 s/mm2with δ=12.0 ms,
Δ=24.2 ms. Spatial resolution was 525×525×600 μm. The total ac-
quisition time was approximately 1 h and 50 min for each imaging
Diffusion data analyses — tractography
1999) described in previous publications (Takahashi et al., 2011, 2012).
The term “streamline” refers to the fact that we connect tractography
pathways using local maximum or maxima. This is true for both DTI
and HARDI. The streamline technique is limited in its ability to resolve
of DTI, as discussed in the DTI paper of Mori et al. (1999). For this reason,
in the current study, we used HARDI, which detects multiple local maxi-
ma on an ODF (orientation distribution function). We used all the local
maxima to produce HARDI tractography pathways, thus enabling us to
identify crossing pathways within a voxel.
Trajectories were propagated by consistently pursuing the orienta-
tion vector of least curvature. We terminated tracking when the angle
between two consecutive orientation vectors was greater than the
given threshold (45°). In many tractography studies, FA values are
used to terminate fibers in the gray matter. In adults, the FA values of
the gray matter are lower than in the white matter. However, because
one of the objectives was to detect fibers in low FA areas (cerebellar
cortex), we used mask (boundary) images of the brains created by
MRIcro (www.sph.sc.edu/comd/rorden/mricro.html) in order to deter-
mine coherenceinthe brain itself and not inthe surrounding immersion
ated from a mean diffusion image of each brain assessment with end-
points constrained to the brain tissue itself. This process is routinely
used to remove skull images or susceptibility-related artifacts around
able alternate method (e.g. Schmahmann et al., 2007; Takahashi et al.,
2012; Vishwas et al., 2010; Wedeen et al., 2008).
Diffusion Toolkit and TrackVis (http://trackvis.org) were used to
reconstruct and visualize tractography pathways. The color-coding of
tractography pathways in Fig. 1 (left panel), Figs. 3, 4A, and 5 are based
on a standard RGB code, applied to the vector between the end-points
In Fig. 2, red identifies the cortico-spinal tract and blue the superior,
green the middle, and yellow the inferior cerebellar peduncle pathways.
In Fig. 6, orange identifies a part of the ponto-cerebellar pathways, blue
identifies pathways running parallel to the long axis of folia, and green
identifies pathways running perpendicular to the blue pathways.
Region of interests (ROIs) and filters for identifying tractography pathways
To improve visualization in the figures, we restricted the number of
tractography pathways displayed in the following ways. For Figs. 1, 4,
and 5, we used sagittal (Figs. 1 and 4) or axial (Fig. 5) slice filters. The
thickness of the slice filters was 3 pixels and 50% of the total pathways
running through the slices were displayed using standard options in
way (pons and spinal cord for the corticospinal pathways, posterior
pons and deep cerebellar white matter for the superior cerebellar pe-
lar peduncle, and posterior pons and cerebellar hemisphere for the
inferior cerebellar peduncle). For Fig. 3, we used only one ROI in the
pons.The positionof theROIis showninthelowerleftcornerof thefig-
ure. For Fig. 6, we used one ROI in the pons to identify yellow or white
pathways and used another ROI in the cerebellar cortex to identify
local cortical pathways. The positions of the ROIs are shown in the
lower left corner of Fig. 6A.
Cerebellar white matter
Cerebellar white matter showed high FA values both in deep, thick
bundles in the medullary core and in the branched thinner laminae
extending out to the folia that constitute the cerebellar cortex
(Fig. 1, asterisk). More than 98% of the total detected white matter
pathways showed FA values ranging from 0.3 to 0.53. The superior
(Fig. 2, dark blue), middle (Fig. 2, green), and inferior (Fig. 2, yellow)
cerebellar peduncles were clearly identified along with the corticospinal
E. Takahashi et al. / NeuroImage 68 (2013) 105–111
tract (Fig. 2, red). The middle cerebellar peduncle conveyed fibers from
the rostral pons to the lateral parts of the cerebellar posterior lobe
(Fig. 3, predominantly light blue) and from the caudal pons to medial
and intermediate regions of the superior aspect of the cerebellar hemi-
sphere (Fig. 3, predominantly green).
ittal plane perpendicular to the folia, the most superficial “layer” showed
high FA values (>0.20) (Fig. 4a), the apparent second layer showed very
low FA values (0–0.05) with less pathways (Fig. 4b), and the apparent
third layer showed medium degrees of FA values (0.05–0.10) (Fig. 4c).
The first and second “layers” were 3–4 mm each, and the third “layer”
was about 2 mm. The thicknesses and the extent of the apparent layers
suggested that these layers result from volume averaging of pathways
from adjacent folia because the spatial resolution of the images is greater
than the thickness of the cortex of the folia (see further discussion in the
section on Limitations and advantages of the current study).
to the folia in the gray matter (Fig. 5a, panels A–C; green pathways) that
were distinct from pathways running in the white matter (Fig. 5b, panel
C; red pathways). The anatomic images upon which the tractography
pathways are superimposed are the mean diffusion images in which
gray matter is seen as white, and white matter is gray (Fig. 5, panel B).
Consequently, the pathways coursing parallel to the folia are evident in
the lighter appearing gray matter, whereas the pathways lying in the
white matter are visible in the gray regions in these mean diffusion im-
ages (Fig. 5, panel B).
We next examined the topographic organization of the different
types of projection pathways. A low magnification view of the right
hemisphere of an adult cerebellum in the axial plane is shown in
Fig. 6A. The yellow box indicates a region of Lobule VIIA, Crus I, which
is magnified in Fig. 6B. Orange pathways were observed perpendicular
to both blue and green pathways. These orange pathways extended
from the pons to the cortical layer of the cerebellum (Fig. 6A and B).
Blue tractography pathways were observed running in a direction paral-
dicular to, but in the same plane as the blue pathways (Fig. 6C and D).
Fig. 6E illustrates the cytology and circuitry of the cerebellar cortex.
Ascending granule cell axons, parallel fibers, basket cell axons, and stel-
late cell axons are situated in the molecular layer. Parallel fibers course
parallel to the long axis of the folia, whereas axons of the cerebellar cor-
tical inhibitory interneurons (stellate cells in the mid and upper reaches
layer) are oriented perpendicular to the long axis of the folia. Therefore,
the blue tractography pathways in Fig. 6A–D likely correspond to groups
of parallel fibers, and the perpendicularly oriented green tractography
pathways in Fig. 6A–D likely represent the axons of stellate and basket
cells. The orange tracks in Fig. 6A–B represent afferents to the cerebellar
cortex including the mossy and climbing fibers, as well as nucleocortical
recurrent collaterals; and efferents from the cerebellar cortex conveyed
by the Purkinje cell axons.
structures within the white matter and cortex of the adult human cere-
bellum post-mortem. In particular, our results demonstrate coherent
fiber track structure likely reflecting the axonal pathways that charac-
terize different cell types within the cerebellar cortex.
Fig. 1. Directional and FA maps of pathways running through a sagittal slice. Cerebellar white matter showed very high FA values both in deep, thick bundles of pathways and also in
branched thinner pathways in the cerebellar cortex (asterisk). A: anterior, P: posterior, D: dorsal, V: ventral, L: left, R: right.
Fig. 2. Identification of white matter pathways. Left panel shows a lateral view, and the right panel shows a medial view of the superior (blue), middle (green), inferior (yellow)
cerebellar peduncles, and the corticospinal tract (red). Small insets show each pathway separately. A: anterior, P: posterior, D: dorsal, V: ventral, L: left, R: right.
E. Takahashi et al. / NeuroImage 68 (2013) 105–111
Topical connections conveyed in the middle cerebellar peduncle
tial pontocerebellar projections in the middle cerebellar peduncle. Fiber
of the cerebellar hemisphere (Fig. 3, mostly light blue), whereas those
from the caudal pons are identified in medial and rostral parts of the
cerebellar hemisphere (Fig. 3, mostly green). This rostral pons to poste-
evident in early myelin stain studies (von Bechterew, 1885) and ana-
tomical investigations (Spitzer and Karplus, 1907; Sunderland, 1940;
see Schmahmann et al., 2004), and is consistent with anatomical, func-
tional imaging, and clinical evidence pointing to a gradient of caudal
pons – anterior lobe of cerebellum sensorimotor representation as op-
posed to a rostral pons – posterior lobe of cerebellum cognitive trend
(Schmahmann and Pandya, 1997; Schmahmann and Sherman, 1998;
Schmahmann et al., 2009; Stoodley and Schmahmann, 2010; Buckner
et al., 2011).
There are several possible reasons why we were able to detect differ-
ential arrangement of pontocerebellar fibers in the middle cerebellar
tion and high signal-to-noise-ratio because artifacts due to movement
and air are minimized. Further, the method of specimen preparation,
using a size-optimized sample container and MR coil, and HARDI
tractography reconstruction further improved the quality of the resulting
tractography pathways (see Takahashi et al., 2010). Given that pathways
in the same bundle (in this case the middle cerebellar peduncle) tend to
run almost in parallel in a small region (in other words, in adjacent
voxels), high-angular resolution is needed to dissociate such pathways.
to detect in vivo, with DTI, or using un-optimized experimental settings.
types of cells within the cerebellar cortex
HARDI tractography detected tangential coherence superficially in
the cerebellar cortex, displaying fibers coursing parallel to the long
axis of the folia. The location and direction of these fibers are consis-
tent with the well-defined features of parallel fibers in the molecular
layer. Crossing these putative parallel fibers were tangential fibers
running perpendicular to the long axis of the folia, consistent with
the axons of stellate cells in the mid-and upper regions of the molec-
ular layer, and of the basket cell axons in the lower part of the molec-
ular layer. These tangential fibers within the cerebellar cortex were
distinct from the fibers linking the cerebellar cortex with the deep
cerebellar nuclei and brainstem.
The ascending granule cell axon divides to form the parallel fibers
which course along the long axis of the cerebellar folium. The branches
of each parallel fiber branch travel for a distance of approximately
3 mmintheratandupto8to10 mminthehuman.Intherat,thebasket
cell axon is approximately 350–400 μm and synapses with the somata
and proximal axons of 9 to 10 Purkinje cells (Palay and Chan-Palay,
Fig. 3. Topograpy of the middle cerebellar peduncle projections. The middle cerebellar peduncle conveyed fibers from the rostral pons to the lateral aspects of the cerebellar pos-
terior lobe (mostly light blue pathways) and from the caudal pons to medial parts of the superior aspect of the cerebellar hemisphere (mostly green pathways). Region of interests
(ROIs) for identifying pathways are shown in the lower left corner. A: anterior, P: posterior, D: dorsal, V: ventral, L: left, R: right.
Fig. 4. Directionally color coded tractography and FA maps of the cerebellar folia with tracts restricted to those remaining within the sagittal slice. A, B. The most superficial layer
showed high FA values (a), the second layer showed very low FA values with fewer tracts identified (b), and the third layer showed medium FA values (c). C. Schematic example of
folia and subfolia showing that tractography pathways are continuous across subfolia likely due to the spatial resolution of the images being 0.525 to 0.600 mm and therefore
averaging across this boundary.
E. Takahashi et al. / NeuroImage 68 (2013) 105–111
1974). In the human data in our study, the tractography pathways sug-
gestive of parallel fibers (blue pathways in Fig. 6) ranged from 3 to
10 mm, and the axons of stellate and basket cells (green pathways in
Fig. 6) ranged from 1 to 10 mm. Tractography pathways represent
mean water diffusivity in a direction within a voxel. It is possible there-
fore, that each tractography path represents multiple, similarly aligned
parallel fibers or axons of interneurons within a voxel, linking different
axon trajectories end-to-end and resulting in a spuriously long course.
Limitations and advantages of the current study
Angle thresholds to terminate tractography fibers are arbitrarily
set in many tractography studies (typically ranging from 35°–60°)
to minimize false positive pathways in some brain areas.
This approach is potentially misleading because an arbitrary setting
that is optimal in one region may be suboptimal in another, resulting in
false fiber tracks. Given the complexity of fiber circuitry and regional
differences in the brain, future studies should investigate methods of
optimizing angle thresholds and other regionally variable tractography
parameters (see also discussion of Takahashi et al., 2010). We were not
able to identify fiber tracks in subfolia as clearly as those in the main
folia, a limitation that may be related to the angle threshold setting, a
oldof 45°for tractographywhichwouldfail to detect those fibers in the
main stem of a foliumwhichturn sharply (almost90°)when enteringa
subfolium. We do not currently use an angle threshold of 90° so as to
limit the likelihood of reconstructing spurious tracks, but in order to
better image pathways in subfolia it may be necessary in future studies
to set different angle thresholds in different brain regions.
We successfully identified tractography pathways likely corre-
sponding to structural coherence arising from groups of parallel fibers
and axons of interneurons in the axial plane (Fig. 5), while in the sag-
ittal plane we observed spurious laminar organization in the cerebel-
lar cortex (Fig. 4A–B). Pathways corresponding to parallel fibers and
axons of interneurons indeed existed in sagittal views, but were
difficult to visualize in that plane using only a sagittal slice filter
(Fig. 5). It was necessary to use a spherical ROI in the cerebellar cortex
in addition to the sagittal slice filter to identify the pathways in the sag-
ittal plane (Fig. 6). Based on the thickness and broad expansion of the
layers parallel to the cerebellar surface, the apparent layers in Fig. 4 do
ture of real fiber pathways across adjacent folia (Fig. 4C). As evident in
the folial structures exemplified in Fig. 4C, the first layer could represent
fibers in the surface of the cerebellum, the second layer could represent
gaps between subfolia belonging to different folia, and the third layer
could represent fibers in deep subfolia adjacent to each other. Given
and coronal directions, all directions have similar degrees of difficulty
when performing tractography.
Coherent tractography pathways likely corresponding to parallel fi-
bers were clearly visualized in axial views (Fig. 5), but the pathways
were not well seen in sagittal views (Fig. 4) mainly because they were
short and running perpendicularly through thesagittal planes. Similarly,
tractography pathways likely corresponding to axons of interneurons
were clearly visualized in sagittal views (Fig. 4, layer a), but not in axial
views (Fig. 5). Therefore it was important to analyze pathways by view-
ing them in all planes, by rotating them in a three dimensional manner,
and often by making another ROI within the cerebellar cortex (Fig. 6).
An advantage of our study is that we have the potential to detect
pathways in subfolia. The spatial resolution of our HARDI imaging was
500–600 μm, while a single subfolium is about 3–4 mm along the short
and the thickness of the white matter in a subfolium is about 2 mm, this
should be sufficient to detect with a spatial resolution of 3–4 voxels.
These studies are ongoing.
We were not privy to the gender of the individuals from whom the
pus callosum and cingulum has been shown in diffusion MRI studies of
the human cerebrum (e.g. Menzler et al., 2011), and additional studies
Fig. 5. Pathways in the cerebellar cortex running through an axial slice. A, B. Views from the top with low intensity (A) and high intensity (B) mean diffusion images. C. A view from
the bottom with high intensity mean diffusion image. Pathways running parallel to folia were detected in the gray matter (a in panel A, B, and C), and they were distinct from path-
ways running in the white matter (b in panel C). A: anterior, P: posterior, D: dorsal, V: ventral, L: left, R: right.
E. Takahashi et al. / NeuroImage 68 (2013) 105–111
withthe gender information will be neededtodeterminewhether gen-
der effects cerebellar white and gray matter pathways as well.
Relevance to future in vivo or validation studies
One of the strengths of our study is the demonstration of the
3-dimensional structure of cerebellar white matter pathways. Further,
to our knowledge this is the first study to depict the 3-dimensional
structure of the cerebellar cortex with this degree of detail. These find-
ings contribute to a more thorough understanding of the organization
of cerebellar circuits and pathways in healthy individuals, and they
have thepotential to shed light on future studies in patients with disor-
ders of the cerebellum.
and at present we cannot identify micro-structural features below this
scale. The observed global view of diffusion coherence in the cerebellar
cortex is nevertheless important as we are able to detect crossing path-
ways in thecerebellar cortex that have not previously been appreciated
using MRI. These results will be relevant for future in vivo studies in pa-
tients, and they also facilitate future immunohistochemical validation
studies (e.g. Xu et al., in press) of the observed imaging findings in post-
mortem cerebella from healthy controls and from patients with cerebel-
lar disease. By building on the findings described here, future in vivo
diffusion tractography studies identifying damage to the cerebellar cor-
tex and white matter have the potential to provide new insights into
ative ataxias and also to the cerebellar component of neuropsychiatric
It is essential to develop a clear picture of the normal patterns and
timing of development of cerebellar pathways and to interpret the role
of white matter pathways in order to more accurately diagnose subtle
disorders of cerebellar connectivity. At this point, the ability to apply
our methods to the in vivo population is limited by motion and possible
spatial resolution, however, progress is being made in that major path-
ways are beginning to be clearly identified (Granziera et al., 2009). The
visualization of the true complexity of a structure is crucial for under-
pathways detected, it will be important for future studies to combine in
vivo MR diffusion tractography with functional MR connectivity.
This work was supported by the Eunice Shriver Kennedy National
Institute of Child Health and Development (NICHD) (R21HD069001)
(ET) and National Institute of Mental Health (R01MH06044) (JDS).
for Biomedical Imaging at the Massachusetts General Hospital, using
ogies, P41RR14075, a P41 Regional Resource supported by the Biomed-
ical Technology Program of the National Center for Research Resources
(NCRR), National Institutes ofHealth. This work alsoinvolvedtheuseof
instrumentation supported by the NCRR Shared Instrumentation Grant
Program (1S10RR023401, 1S10RR019307, and 1S10RR023043) and
High-End Instrumentation Grant Program (S10RR016811).
Buckner, R.L., Krienen, F.M., Castellanos, A., Diaz, J.C., Yeo, B.T., 2011. The organization
of the human cerebellum estimated by intrinsic functional connectivity.
J. Neurophysiol. 106, 2322–2345.
Brodal, P., Bjaalie, J.G., 1997. Salient anatomic features of the cortico-ponto-cerebellar
pathway. Prog. Brain Res. 114, 227–249 (Review).
Granziera, C., Schmahmann, J.D., Hadjikhani, N., Meyer, H., Meuli, R., Wedeen, V.J.,
Krueger, G., 2009. Diffusion spectrum imaging shows the structural basis of
functional cerebellar circuits in the human cerebellum in vivo. PLoS One 4,
Huang, H., Xue, R., Zhang, J., Ren, T., Richards, L., Yarowsky, P., Miller, M.I., Mori, S., 2009.
Anatomical characterization of human fetal brain development with diffusion tensor
magnetic resonance imaging. J. Neurosci. 29, 4263–4273.
Leergaard, T.B., White, N.S., de Crespigny, A., Bolstad, I., D'Arceuil, H., Bjaalie, J.G., Dale,
A.M., 2010. Quantitative histological validation of diffusion MRI fiber orientation
distributions in the rat brain. PLoS One 5 (1), e8595.
Menzler, K., Belke, M., Wehrmann, E., Krakow, K., Lengler, U., Jansen, A., Hamer, H.M.,
Oertel, W.H., Rosenow, F., Knake, S., 2011. Men and women are different: diffusion
tensor imaging reveals sexual dimorphism in the microstructure of the thalamus,
corpus callosum and cingulum. NeuroImage 54, 2557–2562.
Fig. 6. A. A low magnification view of the right hemisphere of an adult cerebellum in the
axial plane. Region of interests (ROIs) for identifying pathways are shown in the lower
left corner. The yellow box shows a region of Crus I, which is magnified in Fig. 6B. B. Blue
tractographypathwayswere observedrunninginadirectionparalleltothe folia inasuper-
blue pathways. Orange pathways were observed running perpendicular to these blue and
green pathways. This pattern of tractography pathways was observed in many regions of
the cerebellum. C–D. The relationship of the blue and green fiber pathways is exemplified
in these two panels. Many of the green fiber pathways were situated within and below
the blue fiber tracks. This spatial relationship suggests that the blue pathways represent
E. An illustration of transverse and longitudinal sectionsof the cerebellum cortex. Note that
allel fibers (blue fibers, blue arrows for example). The longitudinally oriented pathways in
terior, P: posterior, D: dorsal, V: ventral, L: left, R: right.
E. Takahashi et al. / NeuroImage 68 (2013) 105–111
Mori, S., Crain, B.J., Chacko, V.P., van Zjl, P.C., 1999. Three-dimensional tracking of Download full-text
axonal projections in the brain by magnetic resonance imaging. Ann. Neurol.
Palay, S., Chan-Palay, V., 1974. Cerebellar Cortex: Cytology and Organization. Springer-
Verlag, New York.
Saksena, S., Husain, N., Das, V., Pradhan, M., Trivedi, R., Srivastava, S., Malik, G.K., Rathore,
oping human cerebellum with histologic correlation. Int. J. Dev. Neurosci. 26, 705–711.
Schmahmann, J.D. (Ed.), 1997. The Cerebellum and Cognition. Academic Press, San
Schmahmann, J.D., 2007. Cerebellum and spinal cord — principles of development,
anatomical organization, and functional relevance. Chapter In: Brice, A., Pulst,
S. (Eds.), Spinocerebellar Degenerations: The Ataxias and Spastic Paraplegias.
Elsevier, New York, pp. 1–60.
Schmahmann, J.D., 2010. The role of the cerebellum in cognition and emotion: Personal
reflections since 1982 on the dysmetria of thought hypothesis, and its historical
evolution from theory to therapy. Neuropsychol. Rev. 20 (3), 236–260.
Schmahmann, J.D., Pandya, D.N., 1997. The cerebrocerebellar system. In: Schmahmann,
J.D. (Ed.), The Cerebellum and Cognition. : Int. Rev. Neurobiol., 41. Academic Press,
San Diego, pp. 31–60.
Schmahmann, J.D., Sherman, J.C., 1998. The cerebellar cognitive affective syndrome.
Brain 121, 561–579.
Schmahmann, J.D., Doyon, J., Toga, A.W., Petrides, M., Evans, A.C., 2000. MRI Atlas of the
Human Cerebellum. Academic Press, California.
Schmahmann, J.D., Rosene, D.L., Pandya, D.N., 2004. Motor projections to the basis
pontis in rhesus monkey. J. Comp. Neurol. 478, 248–268.
Schmahmann, J.D., Pandya, D.N., Wang, R., Dai, G., D'Arceuil, H.E., de Crespigny, A.J.,
Wedeen, V.J., 2007. Association fibre pathways of the brain: parallel observations
from diffusion spectrum imaging and autoradiography. Brain 130, 630–653.
Schmahmann, J.D., MacMore, J., Vangel, M., 2009. Cerebellar stroke without motor def-
icit: Clinical evidence for motor and non-motor domains within the human cere-
bellum. Neuroscience 162, 852–861.
Spitzer, A., Karplus, J.P., 1907. Über experimentelle Läsionen an der Gehirnbasis. Arbeiten
aus dem Neurologischen Institut, 16, pp. 348–436.
Strick, P.L., Dum, R.P., Fiez, J.A., 2009. Cerebellum and nonmotor function. Annu. Rev.
Neurosci. 32, 413–434.
Stoodley, C.J., Schmahmann, J.D., 2010. Evidence for topographic organization in the
cerebellum of motor control versus cognitive and affective processing. Cortex 46,
Sunderland, S., 1940. The projection of the cerebral cortex on the pons and cerebellum
in the macaque monkey. J. Anat. 74, 201–226.
Takahashi, E., Dai, G., Wang, R., Ohki, K., Rosen, G.D., Galaburda, A., Grant, P.E., Wedeen,
V.J., 2010. Development of cerebral fiber pathways in cats revealed by diffusion
spectrum imaging. NeuroImage 49, 1231–1240.
Takahashi, E., Dai, G., Rosen,G.D., Wang, R.,Ohki, K., Folkerth,R.D.,Galaburda,A., Wedeen,
V.J., Grant, P.E., 2011. Developing neocortex organization and connectivity in cats re-
vealed by direct correlation of diffusion tractography and histology. Cereb. Cortex 21,
Takahashi, E., Folkerth,R.D., Galaburda, A., Grant, P.E., 2012. Emergingcerebral connectivity
in the human fetal brain: an MR tractography study. Cereb. Cortex 22, 455–464.
Tuch, D.S., Reese, T.G., Wiegell, M.R., Wedeen, V.J., 2003. Diffusion MRI of complex neu-
ral architecture. Neuron 40, 885–895.
Vishwas, M., Chitnis, T., Pienaar, R., Healy, B.C., Grant, P.E., 2010. Tract based analysis of
callosal, projection and association pathways in pediatric patients with multiple
sclerosis: a preliminary study. Am. J. Neuroradiol. 31, 121–128.
von Bechterew, W., 1885. Zur Anatomie der Schenkel des Kleinhirns, insbesondere der
Brückenarme. Neurolog. Centralblatt 4, 121–125.
Wedeen, V.J., Wang, R.P., Schmahmann, J.D., Benner, T., Tseng, W.Y., Dai, G., Pandya, D.N.,
Hagmann, P., D'Arceuil, H., de Crespigny, A.J., 2008. Diffusion spectrum magnetic reso-
nance imaging (DSI) tractography of crossing fibers. NeuroImage 41, 1267–1277.
Xu, G., Takahashi, E., Folkerth, R.D., Haynes, R.L., Volpe, J.J., Grant, P.E., Kinney, H.C.,
2012. Radial coherence of diffusion tractography in the cerebral white matter of
the human fetus: Neuroanatomic insights. Cerebral Cortex. [Epub ahead of print]
E. Takahashi et al. / NeuroImage 68 (2013) 105–111