Anatomia, Histologia, Embryologia

ORIGINAL ARTICLE

Fibre Dissection and Sectional Study of the Major Porcine

Cerebral White Matter Tracts
R. Pascalau1* and B. Szabo2,3
Addresses of authors: 1 Iuliu Hatieganu University of Medicine and Pharmacy, 8 Babes Street, 400012 Cluj-Napoca, Romania;
2
Department of Anatomy and Embryology, Iuliu Hatieganu University of Medicine and Pharmacy, 8 Babes Street, 400012 Cluj-Napoca,
Romania;
3
Department of Ophthalmology, Emergency County Hospital, 3-5 Clinicilor Street, 400006 Cluj-Napoca, Romania

*Correspondence: Summary
Tel.: 0040 734 415 295;
fax: 0040 264 597257; White matter anatomy is the basis for numerous applications in neurology,
e-mail: raluca.pascalau@yahoo.ro neurosurgery and fundamental neuroscience. Although the porcine brain is fre-
quently used as experimental model in these fields of research, the description
With 9 figures and 1 table of its white matter is not as thorough as in the human brain or other species.
Thus, the aim of this study is to describe the porcine white matter tracts in a
Received February 2016; accepted for
complex manner. Two stepwise dissection protocols adapted from human anat-
publication June 2017
omy were performed on six adult pig brain hemispheres prepared according to
doi: 10.1111/ahe.12280 the Klingler method. Other four hemispheres were sectioned along section
planes that were chosen similar to the Talairach coordinate system. As a result,
three commissural tracts, seven association tracts and one projection tract were
identified: corpus callosum, fornix, commissura rostralis, the short-association
tracts, fasciculus longitudinalis superior, fasciculus uncinatus, fasciculus longi-
tudinalis inferior, fasciculus occipitofrontalis inferior, cingulum, tractus mamil-
lothalamicus and capsula interna. They were described and illustrated from
multiple points of view, focusing on their trajectory, position, dimensions and
anatomical relations. All in all, we achieved a three-dimensional understanding
of the major tracts. The results are ready to be applied in future imagistic or
experimental studies.

neuroscience does also benefit from the knowledge of

Introduction
The anatomy of the cerebral white matter is nowadays
more and more important due to its applications in neu-
rology, neurosurgery and fundamental neuroscience that
arose together with the development of the Diffusion
Tensor Imaging (DTI) techniques. For example, fractional
anisotropy is an indicator for the evolution of neurologi-
cal diseases such as stroke, multiple sclerosis, Alzheimer’s
disease or epilepsy (Concha et al., 2005; Sbardella et al.,
2013; Gerrish et al., 2014; Sun et al., 2014; Xu et al.,
2014). What is more, tractographic reconstructions are
used for assessment and surgical planning in traumatic
brain injury (Zappal
a et al., 2012; Jang and Seo, 2015),
brain tumours (Bello et al., 2010; Gras-Combe et al.,
2012) and Deep Brain Stimulation (Coenen et al., 2012;
Henderson, 2012; Torres et al., 2014). Fundamental
white matter anatomy especially due to the stimulation
mapping studies that explore the functional connectivity
of the brain (Catani et al., 2005; Fortin et al., 2012; Duf-
fau, 2015; Sarubbo et al., 2015; Zemmoura et al., 2015).
All these applications can be further developed by
experimental research on animal models. It is well known
that the pig (Sus scrofa) is an excellent model in neuro-
science because of the resemblance of its anatomy, distri-
bution of gray and white matter and embryological
development with the human brain (Andersen et al.,
2005; Lind et al., 2007; Sauleau et al., 2009; Radlowski
et al., 2014). Moreover, its dimensions make it more
appropriate than other laboratory animals for imaging
studies or surgical experiments (Holm and West, 1994;
Saikali et al., 2010; Conrad et al., 2014). As a result it is
an alternative to non-human primates in pre-clinical

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378 Anat. Histol. Embryol. 46 (2017) 378–390
R. Pascalau and B. Szabo
safety studies (Ettrup et al., 2010), Deep Brain Stimula-
tion research (Sauleau et al., 2009; Knight et al., 2013),
brain development studies (Radlowski et al., 2014),
pathophysiological studies (Wang et al., 2012) and neuro-
surgical training (Aurich et al., 2014).
In this context, it is worth to explore the porcine white
matter anatomy thoroughly. However, apart from the
internal capsule, corpus callosum and the anterior com-
missure, which were indirectly studied by use of imaging
techniques (Saikali et al., 2010; Dyrby et al., 2007; Yun
et al., 2011; Conrad et al., 2014), the white matter tracts
in the porcine brain are not so well documented in the
scientific literature. Most of the imaging or histological
studies present the white matter of each lobe as a whole
(Watanabe et al., 2001; Wang et al., 2012; Radlowski
et al., 2014). The aim of this study is to provide a
description of the major white matter tracts of the por-
cine brain in a manner that is comparable with the stud-
ies on human white matter (Fernadez-Miranda et al.,
2008; Dini et al., 2013). The Klingler preparation method
(Klingler, 1935; Ludwig and Klingler, 1956; T€ ure et al.,
2000; Martino et al., 2011) is based on the formation of
formaldehyde crystals that separate the fibres so that they
can be dissected in a three-dimensional fashion which is
homologue with virtual tractogaphy (Lawes et al., 2008;
Agrawal et al., 2011). Combining it with standard sec-
tions offers a more complex view and compatibility with
pig brain atlases (Felix et al., 1999; Yun et al., 2011; Con-
rad et al., 2014).
Materials and Methods
Preparation of the specimens
The study was conducted on 10 cerebral hemispheres
coming from adult pigs, which were purchased from a
butcher. They were preserved in 4% formaldehyde solu-
tion for 6 months.
After this period, six hemispheres were prepared for
fibre dissection according to the Klingler method as fol-
lows: the blood vessels and the meningeal layers were
removed under 109–209 magnifying loops. The speci-
mens were immersed in 9% formaldehyde solution and
kept 14 days at 20°C. They then thawed under running
water for several hours.
The other four hemispheres were used for sections.
Fibre dissection
Two stepwise dissection protocols (Fig. 1) previously
adapted from human anatomy by our research group
(Pascalau et al., 2016) were performed each on three
hemispheres. Wooden spatulas and 109–209 magnifying
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White Matter Fibre Dissection and Sections in Pig
loops were used to remove the grey matter structures (the
cortex, the thalamus and some of the basal ganglia, which
become very friable after freezing) and to separate the
adjacent white matter tracts according to the cleavage
planes that have been created by the ice crystals during
the preparation. The surgical scalpel was used just for
two cuts in the entire dissection to respect as much as
possible the anatomical relations of the white matter
tracts. At each step of the dissections, digital photographs
from the same distance were taken and at some stages
diluted Indian ink was spread on the specimen to
enhance the visualization of the trajectory of the fibres.
The mediolateral fibre dissection protocol has six steps.
The first step consists of identifying the sulci and gyri on
the medial surface of the cerebral hemisphere and of
removing the cerebral cortex beginning from the depth of
the sulcus to the top of the gyrus. At the end of this step,
the short-association fibres and the cingulum can be visu-
alized. The second step is the exposure of the white mat-
ter tracts that are located under the ependymal layer of
the third ventricle, namely, the anterior pillar of the for-
nix, tractus mamillothalamicus and tractus mamil-
lotegmentalis, which will be followed to their projections
in the nucleus thalamic anterior and lamina tecti respec-
tively. Commissura rostralis is also dissected in this step
of the dissection. In the third step, the uppermost cal-
losal fibres are revealed by removing the cingulum, the
short-association fibres and some of the projection fibres
that intersect corpus callosum. In the fourth step of the
dissection, the body and rostral horn of the lateral ventri-
cle are exposed by eliminating corpus callosum. The fifth
step aims to reach corona radiata. It requires the removal
of the head of nucleus caudatus with the wooden spatula
and the excision of the anterior part of the fornix by use
of a horizontal cut at the level of the pulvinar thalami.
Finally, the sixth step of the dissection is the exposure of
the medial surface of the capsula interna which is done
by taking off the thalamus. A horizontal cut is made at
the level of the hypothalamus. The spatula is inserted in
the depth of the cut and the thalamus is pooled upwards
and extracted together with corona radiata. The capsula
interna fibres can be followed from the cortical surface to
the cerebral peduncle.
The other fibre dissection protocol begins from the lat-
eral surface of the brain and includes four steps. In the
first step, the sulci and gyri on the lateral surface are
identified and the cortex is removed so that the short-
association fibres can be seen. The second step consists
of the exposure of capsula externa and the most superfi-
cial long-association tracts, such as fasciculus longitudi-
nalis superior, fasciculus longitudinalis inferior and
fasciculus uncinatus, which are situated just underneath
the short-association fibres. Claustrum and capsula
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White Matter Fibre Dissection and Sections in Pig
Fig. 1. Fibre dissection protocols. (see explanation in text).
extrema cannot be separated from the short-association
fibres. Capsula externa can be identified as a sheet of
white matter located dorsally to fissura sylvia. It must be
removed in the third step of the dissection to expose
nucleus lentiformis and fasciculus occipitofrontalis infe-
rior. The fourth step begins with the removal of nucleus
lentiformis which results in revealing the lateral surface of
the genu of capsula interna. Its fibres are then followed to
the cortical surface.
Sections
To be in concordance with some MRI or histological stud-
ies of the porcine brain (Felix et al., 1999; Watanabe et al.,
2001; Saikali et al., 2010; Yun et al., 2011; Conrad et al.,
2014), the section planes were chosen similar to the
Talairach coordinate system (Talairach and Tournoux,
1988). The axial sections were parallel to the anterior
commissure–posterior commissure line (AC–PC line), the
coronal sections were parallel to a plane which is perpen-
dicular to the AC–PC line and the sagittal sections were
parallel to the midsagittal plane. The section thickness was
4 mm. By use of a milimetric scale, each section could be
380
R. Pascalau and B. Szabo
located in the Talairach coordinate system, considering
that the PC in the midsagittal section is the origin of the
sistem, the AC-PC line is the Ox axis, the perpendicular
vertical line passing through the PC is the Oy axis and the
left–right axis with the origin in PC is Oz.
The research protocol and the use of animal specimens
were approved by the Ethics Committee of “Iuliu Hație-
ganu” University of Medicine and Pharmacy, Cluj-
Napoca.
Results
White matter tracts are groups of neuronal axons which
have parallel trajectories forming fascicles within the sub-
cortical space. Their aim is to connect different functional
areas of the brain and to create the brain functional net-
work. According to the structures connected, white mat-
ter tracts can be classified in: commissural fibres,
connecting homologue cortical areas in the two hemi-
spheres; association fibres, connecting different cortical or
subcortical areas within the same hemisphere and projec-
tion fibres which connect cortical with subcortical struc-
tures. As a general rule, commissural, association and
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R. Pascalau and B. Szabo White Matter Fibre Dissection and Sections in Pig

Fig. 2. Corpus callosum and fornix. (a) Mid-

sagittal section. (b) Mediolateral fibre dissec-
tion (third step, inked). (c) Coronal section
(x = 4 mm). (d) Transversal section
(y = 8 mm). (e) Mediolateral fibre dissection
(fourth step). b, body of corpus callosum; c,
cingulum; cc, corpus callosum; ec, capsula
externa; f, fornix; g, genu of corpus callosum;
hc, hippocampus; ic, capsula interna; nc,
nucleus caudatus; nl, nucleus lentiformis; r,
rostrum of corpus callosum; s, splenium of
corpus callosum; t, thalamus.

projection tracts can be recognized by their transversal,
sagittal and longitudinal directions respectively.
Commissural tracts
Corpus callosum
Being the largest white matter tract in the porcine brain,
corpus callosum is a symmetrical dorsally convex struc-
ture situated on the midline above the lateral ventricles
(Fig. 2). On the midsagittal section it has a “C” shape,
with 36.7 mm length and maximum width of 2.9 mm,
that can be divided rostrocaudally in rostrum, genu,
body and splenium. This shape also appears on sagittal
sections from z = 12 mm to z = 12 mm. Callosal radi-
ations connecting homologue frontal, parietal, occipital
and temporal cortical areas are best visualized in
coronal sections from x = 12 mm to x = 16 mm and
in the third step of the mediolateral fibre dissection.
The radiations of splenium and genu are best visible on
transversal sections (y = 0–8 mm). The dorsal surface of
corpus callosum has relations with cingulum and with
the projection fibres that intersect it in their way to the
cortex. Its ventral surface is the external wall of the lat-
eral ventricle and it has contact with nucleus caudatus
and fornix.
Fornix
Fornix is a commissural tract belonging to the limbic sys-
tem (Fig. 2). Its body can be seen in the fourth step of
the mediolateral fibre dissection as a triangular mass
placed in a transversal plane above the thalamus. On
frontal sections (x = 8 to 8 mm) it looks like a

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White Matter Fibre Dissection and Sections in Pig
transversal band with a maximum width of 3.2 mm and
on sagittal sections (z = 4 to 4) it has an arcuate shape.
It is continued rostrally by the 11-mm-long and 1-mm-
wide anterior pillars which descend to corpora mamillar-
ia. They can be exposed in the second step of the medio-
lateral fibre dissection. The caudal extensions of the body
are crura fornicis which disperse in the white matter of
the hippocampus. Important anatomical relations of for-
nix are the thalamus, which it surrounds, the trunk of
commissura rostralis, which crosses the anterior pillars
and corpus callosum.
Commissura rostralis
Commissura rostralis is an “H”-shaped structure connect-
ing the two olfactive bulbs and temporal cortices (Fig. 3).
Its trunk appears on the midsagittal section as a circle
with 1.5 mm diameter situated at (x = 20 mm,
y = 0 mm) and on the x = 20 mm frontal section it
looks like a 38-mm-long band. The rostral radiations
diverge like a fan with 38° aperture to the olfactive bulb
and they can be revealed in the second step of the medio-
lateral fibre dissection and on z = 4 mm sagittal and
y = 0 mm transversal sections. On the other hand, the
posterior radiations cannot be distinguished from the
long-association tracts that run within the temporal lobe.
Fig. 3. Commissura rostralis. (a) Coronal section (x = 20 mm). (b) Transversal section (y = 0 mm). (c) Sagittal section (z = 4 mm). (d) Mediolat-
eral fibre dissection (second step). c, cingulum; cc, corpus callosum; ec, capsula externa; f, fornix; ic, capsula interna; na, nucleus accumbens; nc,
nucleus caudatus; nl, nucleus lentiformis; oc, chiasma opticum; rc, commissura rostralis; t, thalamus.
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R. Pascalau and B. Szabo
The anatomical relations of commissura rostralis from
medial to lateral are as follows: the anterior pillar of for-
nix, head of nucleus caudatus, the anterior limb of cap-
sula interna and nucleus accumbens.
Association tracts
Short-association tracts
Also known as arcuate fibres, the short-association tracts
connect adjacent gyri (Fig. 4). They are situated the most
superficial, just underneath the cortex of the sulci. As a
result, they represent the first white matter fibres encoun-
tered in the stepwise fibre dissections. Frontal sections
show that these “U”-shaped fibre bundles run perpendic-
ular to the length of each sulcus and they can be used to
determine the Gyrification Index (GI) defined as the ratio
between the total outer cortical surface and the superfi-
cially exposed part of the cortex (Table 1). The length of
the short-association tracts increases with the GI. The
longest are the ones of sulcus rhinalis: 25.16 mm and sul-
cus cruciatus: 17.33 mm. At this level (x = 4–16 mm) GI
is also the highest. The projection fibres and the most
superficial long-association fibres such as cingulum, fasci-
culus longitudinalis superior and inferior are the most
important anatomical relations of the arcuate fibres.
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R. Pascalau and B. Szabo White Matter Fibre Dissection and Sections in Pig

Fig. 4. Short-association tracts. (a) Medial sur-

face of the brain. (b) Mediolateral fibre dis-
section (first step). (c) Lateral surface of the
brain. (d) Lateromedial fibre dissection (first
step). (e) Coronal section (x = 28 mm). (f)
Coronal section (x = 8 mm). bo, bulbus
olfactorius; c, cingulum; fs, fissura sylvia; gc,
gyrus cinguli; gco, gyrus coronalis; gem, gyrus
ectomarginalis; ges, gyrus ectosylvius; gfs,
gyrus frontalis superior; gm, gyrus marginalis;
gs, gyrus sigmoideus; gs, gyrus sylvius; sa, sul-
cus ansatus; sc, sulcus cruciatus; scg, sulcus
cynguli; sd, sulcus diagonalis; se, sulcus ecto-
sylvius caudalis; sl, sulcus lateralis; sn, sulcus
naris; sp, sulcus pre-sylvius; sr, sulcus rhinalis;
ss, sulcus splenialis; ssc, sulcus suprasylvius
caudalis; ssg, sulcus sagittalis; ssr, sulcus
suprasylvius rostralis.

Table 1. Gyrification index (GI) at different frontal planes transversal sections (y = 12–16 mm), whereas on the
coronal sections (x = 8 to 24 mm) it appears like a
X (mm) Gyrification index
round or flat structure. Before entering the parietal lobe
16 1.91 it makes a 143° genu. Fasciculus longitudinalis superior
12 1.6 is related to the short-association fibres superficially and
8 1.97 with the capsula interna–corpus callosum intersection
4 1.74
area deeply.
4 2
16 2.27
24 1.77 Capsula externa
28 1.68 Capsula externa is a 2.34-mm-wide layer of white matter
Average 1.8675 located between claustrum, laterally, and nucleus len-
Standard deviation 0.215257 tiformis, medially. It is traversed by two long-association
tracts: fasciculus uncinatus and fasciculus longitudinalis
inferior (Fig. 5). On transversal sections (y = 4 to
12 mm) it looks like a 21.2-mm-long band and on coro-
Fasciculus longitudinalis superior nal sections (x = 0–24 mm) it is “V” shaped, forming a
The long-association tract connecting the frontal and triangle with capsula interna around nucleus lentiformis.
parietal lobes is fasciculus longitudinalis superior The anatomical relations of the medial surface of capsula
(Fig. 5). It is a 62.27-mm-long and 4.4-mm-wide band- externa are capsula interna, nucleus lentiformis and fasci-
like fibre bundle, situated just underneath the short- culus occipitofrontalis inferior, in dorsoventral order. Its
association tracts of the dorsolateral region of the brain lateral surface has contact with claustrum and some arcu-
so that it can be reached in the second step of the ate fibres. Because claustrum is removed together with
lateromedial fibre dissection. This shape can be recog- the arcuate fibres of gyrus ectosylvius, gyrus sylvius and
nized on sagittal (z = 12 mm to z = 16 mm) and sulcus suprasylvius, which represent capsula extrema,

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White Matter Fibre Dissection and Sections in Pig R. Pascalau and B. Szabo

Fig. 5. Fasciculus longitudinalis superior, capsula externa and fasciculus occipitofrontalis inferior. (a) Midsagittal section. (b) Sagittal section
(z = 12 mm). (c) Transversal section (y = 12 mm). (d) Coronal section (x = 8 mm). (e) Coronal section (x = 20 mm). (f) Transversal section
(y = 4 mm). (g) Lateromedial fibre dissection (second step). (h) Lateromedial fibre dissection (third step). c, cingulum; cc, corpus callosum; ec,
capsula externa; f, fornix; fli, fasciculus longitudinalis inferior; fls, fasciculus longitudinalis superior; foi, fasciculus occipitofrontalis inferior; fu, fasci-
culus uncinatus; hc, hippocampus; ic, capsula interna; lv, lateral ventricle; m, mesencephalon; nc, nucleus caudatus; nl, nucleus lentiformis; co,
chiasma opticum; rc, commissura rostralis; ro, radiatio optica; t, thalamus.

capsula externa is revealed at the second step of the
lateromedial fibre dissection. At this stage of the dissec-
tion it plays the role of a landmark for the trunks of fas-
ciculus uncinatus and fasciculus longitudinalis inferior.
Their radiations must be followed from here to the corti-
cal surface by further removal of the short-association
fibres.
Fasciculus uncinatus runs from the frontal pole to the
temporal pole through capsula externa, in the depth of
sulcus rhinalis. It has the shape of letter “U” and a total
length of 51.9 mm and maximum width of 8.57 mm.
The arms of fasciculus uncinatus form an angle of 55°.
Its trunk or its arms can be found on sagittal (z = 8–
12 mm), transversal (y = 8 to 8 mm) or frontal
(x = 16–24 mm) sections in the inferior part of capsula
externa, lateral to fasciculus occipitofrontalis inferior and
rostral to fasciculus longitudinalis inferior.
Fasciculus longitudinalis inferior connects the temporal
pole with the occipital lobe. It is a 41.9-mm-long and
5.71-mm-wide band which passes through capsula externa
and underneath the temporal and occipital arcuate fibres.
It appears on sagittal and transversal sections taken at the
same levels as fasciculus uncinatus and on the frontal sec-
tions taken more caudally (x = 16 to 8 mm). Its

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384 Anat. Histol. Embryol. 46 (2017) 378–390
R. Pascalau and B. Szabo
temporal radiations have contact with the temporal arm
of fasciculus uncinatus and its occipital radiations have
medial relation with fasciculus occipitofrontalis inferior.
Fasciculus occipitofrontalis inferior
If fasciculus uncinatus and fasciculus longitudinalis infe-
rior provide indirect fronto-occipital connections, deep to
them there is a direct long-association tract, fasciculus
occipitofrontalis inferior (Fig. 5). It can be reached in the
third step of the lateromedial fibre dissection using as
landmark nucleus lentiformis, which is situated just dor-
sal to its trunk, from which the frontal and occipital radi-
ations diverge forming a 133° angle and a total length of
5.4 mm. On transversal (y = 4 to 4 mm) and sagittal
(z = 12 mm) sections it looks like a 2.2-mm-wide and
8.2-mm-high band, whereas on coronal sections
(x = 16 to 24 mm) it has various heights. Travelling a
long distance in the brain, it has numerous anatomical
relations such as commissura rostralis and capsula interna
(especially radiatio optica) medially, nucleus lentiformis
dorsally and fasciculus uncinatus and fasciculus longitudi-
nalis inferior laterally.
Cingulum
Cingulum is a limbic long-association tract connecting
the subcallosal frontal lobe with lobus piriformis (Fig. 6).
Fig. 6. Cingulum. (a) Coronal section (x = 8 mm). (b) Mediolateral fibre dissection (first step). (c) Sagittal section (z = 4 mm). (d) Mediolateral
fibre dissection (first step, dorsocaudal view). c, cingulum; cc, corpus callosum; cr, corona radiata; f, fornix; ht, hypothalamus; ic, capsula interna;
lv, lateral ventricle; rc, commissura rostralis; sc, colliculus quadrigeminus rostralis; t, thalamus; 3v, third ventricle.
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White Matter Fibre Dissection and Sections in Pig
It surrounds corpus callosum and fornix and is located
just underneath the cortex of gyrus cinguli and lobus pir-
iformis so it can be exposed in the first step of the medi-
olateral fibre dissection. On the first sagittal section
(z = 4 mm) it follows the external contour of corpus
callosum and some of its frontal, parietal and occipital
radiations are visible. They give the cingulum a triangular
shape in the frontal sections (x = 8 to 20 mm). It has
maximum width of 1.8 mm and a total length of
90.9 mm. It has anatomical relations with the short-asso-
ciation fibres of the medial and basal surfaces of the
brain, with the dorsal surface of corpus callosum and
with crus fornicis.
Tractus mamillothalamicus
There is a limbic white matter tract accessible under
the ependymal layer of the third ventricle (Fig. 7). It
can be revealed in the second step of the mediolateral
fibre dissection. Tractus mamillothalamicus is a cylindri-
cal 1-mm-thick, 13-mm-long tract running from cor-
pora mamillaria to the anterior thalamic nuclei. It
makes a 27° angle with the anterior pillar of fornix, a
80° angle with tractus mamillotegmentalis and a 24°
angle with the Oy axis. It appears on the y = 4 mm
transversal and x = 16 mm frontal sections. Its
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White Matter Fibre Dissection and Sections in Pig
anatomical relations are the subthalamic nuclei and the
third ventricle.
Projection tracts
Capsula interna
Most of the corticonuclear, corticospinal thalamocortical
(corona radiata) projection tracts and some interstriate
fibres are concentrated in the centre of each cerebral
hemisphere, in capsula interna (Fig. 8). It has the shape
of an open fan continued by the cerebral peduncle and it
represents the final target for both lateromedial and
mediolateral fibre dissection techniques. On frontal sec-
tions (x = 8 to 16 mm) it can be recognized as an 5.62-
mm-wide and 34.37-mm-long oblique band forming an
19° angle of with the Oy axis. It is the medial edge of a
triangle formed together with capsula externa around
nucleus lentiformis. The transversal sections (y = 4 to
8 mm) show its well-known “V” shape which can be
divided into anterior limb, genu and posterior limb
which continues with the retrolenticular portion, namely,
radiatio optica. It has a total length of 31.25 mm, maxi-
mum width of 5 mm and the limbs form a 126° angle at
the level of genu so that they act as two edges of a trian-
gle around nucleus lentiformis whose third edge is cap-
sula externa. Sagittal sections (z = 4–8 mm) reveal
capsula interna as a white matter trapezoid surrounded
by the basal ganglia.
If we consider capsula interna as an open fan, we
should nominate the anatomical relations of its medial
surface, lateral surface, rostral edge, caudal edge, dorsal
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R. Pascalau and B. Szabo
Fig 7. Tractus mamillothalamicus. (a)
Transversal section (y = 4 mm). (b) Coronal
section (x = 16 mm). (c) Mediolateral fibre
dissection (second step). cc, corpus callosum;
cm, corpus mamillare; ec, capsula externa; f,
fornix; ic, capsula interna; lv, lateral ventricle;
na, nucleus accumbens; nc, nucleus caudatus;
nl, nucleus lentiformis; co, chiasma opticum;
rc, commissura rostralis; t, thalamus; tmtg,
tractus mamillotegmentalis; tmth, tractus
mamillothalamicus.
edge and root. The relations of the medial surface are
nucleus caudatus rostrally, thalamus and the occipital
horn of the lateral ventricle for the retrolenticular por-
tion. The lateral surface has contacts with nucleus len-
tiformis, fasciculus occipitofrontalis inferior, capsula
externa and fasciculus longitudinalis superior. The rostral
edge is crossed by commissura rostralis and embraced by
nucleus accumbens. The caudal edge is located between
the arcuate fibres of the occipital lobe. In order to reach
the dorsal edge of capsula interna, namely, the cortical
surface, the projection fibres have to intersect the
transversal fibres of corpus callosum at the level of cap-
sula interna–corpus callosum intersection area, which can
be felt in during the fibre dissection as a very firm white
matter region. Within each gyrus, the projection fibres
are closely related to the short-association fibres. The root
of the fan is represented by the cerebral peduncle which
comes in contact with the nuclei and fascicles of the
brainstem.
Discussion
The multiple approaches in which this study presents the
white matter tracts of the porcine brain are aimed to pro-
vide a three-dimensional understanding of the porcine
white matter anatomy, which is summarized in Fig. 9.
This scheme was created based on the data included in
the Results, such as the order in which the tracts are
exposed in the stepwise fibre dissections and their posi-
tion on the standard sections. The Klingler method has
the advantage of preserving the spatial anatomy (Dini
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Anat. Histol. Embryol. 46 (2017) 378–390
R. Pascalau and B. Szabo White Matter Fibre Dissection and Sections in Pig

Fig. 8. Capsula interna. (a) Coronal section (x = 8 mm). (b) Transversal section (y = 4 mm). (c) Lateromedial fibre dissection (fourth step, inked).
(d) Sagittal section (z = 8 mm). (e) Mediolateral fibre dissection (sixth step, inked). al, anterior limb of capsula interna; c, cingulum; cc, corpus
callosum; cp, cerebral peduncle; ec, capsula externa; f, fornix; g, genu of capsula interna; ht, hypothalamus; ic, capsula interna; lv, lateral ventri-
cle; nc, nucleus caudatus; nl, nucleus lentiformis; pl, posterior limb of capsula interna; t, thalamus; 3v, third ventricle.

et al., 2013; Yaman et al., 2014; Zemmoura et al., 2015)
because it does not require force or cuts. It has also been
proved to be reproducible with some adjustments in ani-
mal anatomy (Yaman et al., 2014; Pascalau et al., 2016).
On the other hand, the sectional study gives a lot of
details about the anatomical relations, which are extre-
mely important. What is more, the association between
sections integrated in the coordinate system and fibre dis-
section images of the same tract could be the basis for
future fibre tracking studies, provided that the Region Of
Interest (ROI) selection is done on sectional images and
that the anatomical knowledge plays a crucial role in this
process (Mori and van Zijl, 2002; Catani, 2007; Leclercq
et al., 2011).
Three-dimensional anatomy plays a very important role
in the research applications of the porcine brain. This is
why studies based on sectional imaging always try to inte-
grate the data in three-dimensional reconstructions
(Watanabe et al., 2001; Saikali et al., 2010; Conrad et al.,
2012, 2014). However, white matter tracts are not indi-
vidually represented in these reconstructions except for
corpus callosum and fornix. There is also no DTI study
on the porcine brain that reconstructs all the tracts such
as in the case of the human, canine or feline brain
(Leclercq et al., 2011; Jacqmot et al., 2013; Anaya Garcia
et al., 2015). As far as we know, only some corpus callo-
sum and capsula interna are used as ROI in the studies
reported in the literature (Dyrby et al., 2007; Wang et al.,
2012; Radlowski et al., 2014).
As a result, some of the white matter tracts illustrated
in this study could not have been compared with other
images of the pig brain so that they were only compared
with fibre dissection or DTI images from other species
(Jacqmot et al., 2013; Yaman et al., 2014; Anaya Garcia
et al., 2015; Pascalau et al., 2016). Comissura rostralis,
tractus mamillothalamicus and long-association tracts like

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Anat. Histol. Embryol. 46 (2017) 378–390 387
White Matter Fibre Dissection and Sections in Pig
Fig. 9. Three-dimensional scheme of the major porcine white matter
tracts in lateromedial view. c, cingulum; cc, corpus callosum; cm, cor-
pus mamillare; cr, commissura rostralis; f, fornix; fli, fasciculus longitu-
dinalis inferior; fls, fasciculus longitudinalis superior; foi, fasciculus
occipitofrontalis inferior; fu, fasciculus uncinatus; hc, hippocampus; ic,
capsula interna; nc, nucleus caudatus; nl, nucleus lentiformis; t, thala-
mus; tmth, tractus mamillothalamicus.
fasciculus longitudinalis superior, fasciculus uncinatus,
fascicuus longitudinalis inferior and cingulum have very
similar shape and position with the equine, bovine,
canine and feline ones. Fasciculus occipitofrontalis infe-
rior was more similar to the bovine and canine ones and
less to the feline and equine ones. As for fasciculus occip-
itofrontalis superior, which was identified in the canine
and feline brains (Jacqmot et al., 2013; Anaya Garcia
et al., 2015; Pascalau et al., 2016), it has not been
encountered in the porcine brain, which resembles it to
the equine and bovine brains (Yaman et al., 2014; Pas-
calau et al., 2016). Capsula interna, corpus callosum and
fornix are comparable to the aforementioned species as
well as with the porcine brain images from the literature
(Dyrby et al., 2007; Saikali et al., 2010). In a general view,
the major white matter tracts have similar organization
with the human ones (Fernadez-Miranda et al., 2008)
except for fasciculus longitudinalis superior which does
not have the temporal projections of its human homo-
logue as a consequence of the more pronounced caudal
expansion and curvature of the primate telencephalon
compared to the porcine one (Lind et al., 2007).
Some limitations to our study could be the use of mid-
line-sectioned hemispheres, which interfered with the
study of the commissural tracts, the thickness of the sec-
tions (4 mm) affecting the precision of the tracts localiza-
tion and the known limitations of the Klingler method
(T€ure et al., 2000; Martino et al., 2011): the risk of fibre
rupture and the impossibility to separate some adjacent
tracts (this was the case of the posterior radiations of
commissura rostralis).
388
R. Pascalau and B. Szabo
Nevertheless, our results and especially our stepwise
dissection methods should also be used in teaching or
training purposes because, in spite of the long learning
curve, the experience of fibre dissection correlated with
brain sectioning enriches the anatomical spatial vision
and the dissecting skills (Martino et al., 2011). The Klin-
gler method is nowadays very popular among neurosur-
geons and trainees (Martino et al., 2011; Yaman et al.,
2014; Zemmoura et al., 2015), but the access to human
brains is limited, whereas the pig brain is very accessible
(Ettrup et al., 2010). Thus, porcine brain fibre dissection
seems like a good alternative.
Conclusions
The major porcine commissural, association and projec-
tion tracts were described in a multiapproach fashion in
order to fulfil the lack of thorough descriptions of them
in the literature in comparison to human anatomy, pro-
vided that the porcine brain is frequently used as animal
model for neuroscience research which aims to have
applications in human biology or medicine. Two stepwise
fibre dissection protocols were used for the exposure of
the fibre bundles in their entire length and standard sec-
tions integrated in a coordinate system were performed
to determine their spatial localization and their dimen-
sions. Special attention was given to the anatomical rela-
tions of each tract.
As a result, a three-dimensional view of the major
tracts was achieved and it was schematically illustrated.
The results are comparable with the literature on pig or
other species and they are ready to be applied in future
imagistic or experimental studies.
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