THE JOURNAL OF COMPARATIVE NEUROLOGY 303:489-511 (1991)

Trigeminal Primary Afferent Projections to

Non-TrigemindAreas of the Rat Central
-
Nervous System
CARL F. MARFURT AND DONNA M. RGTCHERT
Northwest Center for Medical Education, Indiana University School of Medicine,
Gary, Indiana 46408
ABSTRACT
The central projections of rat trigeminal primary afferent neurons to various non-
trigeminal areas of the central nervous system were examined by labeling the fibers with
wheat germ agglutinin-horseradish peroxidase (WGA-HRP) transported anterogradely from
the trigeminal ganglion. This technique produced a clear and comprehensive picture of
trigeminal primary afferent connectivity that was in many ways superior to that which may be
obtained by using degeneration, autoradiography, cobalt labeling, or HRP transganglionic
transport techniques. Strong terminal labeling was observed in all four rostrocaudal subdivi-
sions of the trigeminal brainstem nuclear complex, as well as in the dorsal horn of the cervical
spinal cord bilaterally, numerous brainstem nuclei, and in the cerebellum. Labeling in the
ipsilateral dorsal horn of the cervical spinal cord was very dense at C1, moderately dense at C2
and C3, and sparse at (2447. Numerous fibers crossed the midline in the medulla and upper
cervical spinal cord and terminated in the contralateral pars caudalis and dorsal horn of the
spinal cord from Cl-C5. The latter axons terminated most heavily in the mandibular and
ophthalmic regions of the contralateral side. Extremely dense terminal labeling was observed in
the ipsilateral paratrigeminal nucleus and the nucleus of the solitary tract, moderate labeling
was seen in the supratrigeminal nucleus and in the dorsal reticular formation, and small
numbers of fibers were observed in the cuneate, trigeminal motor, lateral and superior
vestibular nuclei, and in the cerebellum. The latter fibers entered the cerebellum in the superior
cerebellar peduncle and projected to the posterior and anterior lobes as well as to the interposed
and lateral deep cerebellar nuclei. Most projections in this study originated from fibers in the
dorsal part of the spinal tract of V, suggesting a predominantly mandibular origin for these
fibers. Projections from the ophthalmic and maxillary divisions, in contrast, were directed
mainly to the cervical spinal cord bilaterally, to contralateral pars caudalis, and to certain areas
of the reticular formation. In conclusion, this study has demonstrated that somatosensory
information from the head and face may be transmitted directly to widespread and functionally
heterogeneous areas of the rat central nervous system, including the spinal cord dorsal horn,
numerous brainstem nuclei, and the cerebellum. These projections may play important roles in
trigeminospinal reflexes (dorsal horn of spinal cord Cl-C7), fusion of the sensory maps of the
right and left sides of the head (contralateral projections), trigeminovisceral integration
(paratrigeminal and solitary), control and integration of oral motor behavior (supratrigeminal
and motor V), orofacial reflexes (reticular formation), and the coordination and stabilization of
head posture and gaze (cuneate, vestibular, and cerebellum).
Key words: Trigeminal nerve, trigeminal ganglion, somatosensory systems, wheat germ
agglutinin-horseradish peroxidase
Sensory information from the head and face is transmit-
ted into the brainstem by the trigeminal nerve. Most
trigeminal primary afferent fibers terminate in the ipsilat-
era1 trigeminal brainstem nuclear complex (TBNC), and
the somatotopical organization and electrophysiological
characteristics of this projection have been well described
(Nord, 67; Kruger, 71; Darian-Smith, 73; Panneton and
Burton, 81; Marfurt, 81; Arvidsson, 82; J acquin et al.,
83b; Shigenaga et al., 86a,b). Other trigeminal primary
Accepted September 26,1990.
o 1991 WILEY-LISS, INC.
490 C.F. MARFURT AND D.M. RAJCHERT
Abbreviations
aP
Cl-C7
coch
comX
Cr 1
Cr2
cun
dm X
dv
ecn
gr
I-v
IV
icp
inter
lat
lv
mes V
mV, motor V
cc
area postrema
cervical spinal cord segments C1-C7
central canal
cochlear nucleus
commissural nucleus of X
crus 1 of the ansiform lobule
crus 2 of the ansiformlobule
cuneate nucleus
dorsal motor nucleus of X
descending (inferior) vestibular nucleus
external cuneate nucleus
gracile nucleus
laminae I-V of pars caudalis and spinal cord dorsal horn
fourth ventricle
inferior cerebellar peduncle
interposed cerebellar nucleus
lateral (dentate) cerebellar nucleus
lateral vestibular nucleus
mesencephalic nucleus of V
trigeminal motor nucleus
afferent fibers, however, have been reported to project to
the spinal cord bilaterally, the contralateral TBNC, various
brainstem nuclei, and the cerebellum. Unfortunately, the
latter projections of the trigeminal ganglion have been less
extensively investigated and much of the available informa-
tion is superficial and occasionally conflicting.
I t appears well established that trigeminal primary af-
ferent fibers project in significant numbers to the ipsilateral
dorsal horn of C1 (indeed, this region is regarded by many
workers as a caudal extension of the TBNC); however, the
extent to which trigeminal primary afferents project to
more caudal levels of the spinal cord is less certain. Whereas
most early studies described projections only to the level of
C2 or C3 (Torvik, '56; Kerr, '631, the results of two recent
horseradish peroxidase tracing studies suggest that some
fibers may descend as far as C6 or C7, at least in the rat
(J acquin et al., '83b; Pfaller and Arvidsson, '88).
The presence and extent of trigeminal primary afferent
projections to the contralateral TBNC and spinal cord have
also been subjects of some controversy. The existence of a
crossed trigeminal input to the dorsal horn of C1 and C2
has been well substantiated by numerous workers (Torvik,
'56; Clarke and Bowsher, '62; Matesz, '83; J acquin et al.,
'83b; Pfaller and Arvidsson, '88; J acquin et al., '90);
however, reports of contralateral projections to the trigemi-
nal main sensory nucleus, (Torvik, '561, pars interpolaris
(Westrum et al., '76), and pars caudalis (J acquin et al., '82,
'83b) remain largely unconfirmed.
Trigerninal primary afferent inputs to various "non-
trigeminal" (extratrigeminal) nuclei' of the brainstem have
also been described by several workers, including promi-
nent projections to the nucleus of the solitary tract (Torvik,
'56; Kerr, '63; Kruger et al., '77; Beckstead and Norgren,
'79; J acquin et al., '82; Pfaller and Arvidsson, '881, para-
trigeminal nucleus (Pfaller and Arvidsson, '88; J acquin et
al., '881, and a more modest projection to the reticular
formation (Torvik, '56; J acquin et al., '82; Matesz, '83;
Pfaller and Arvidsson, '88). Other areas that have been
reported to receive lesser amounts of trigeminal primary
afferent input include the supratrigeminal (J acquin et al.,
'82, '83b; Shigenaga et al., '86b; Takemura et al., '87),
' For the purpose of this paper, the term, "non-trigeminal" nucleus refers
to any CNS nucleus exclusive of the trigeminal brainstem nuclear complex.
MSN
mv
para V
pcRF
PM
pyr decus
RF
Scp
:&
sol
sptrV
supra V
VII
sv
VII n, VII nerve
VII genu
XI1
2-5
trigeminal main sensory nucleus
medial vestibular nucleus
paratrigeminal nucleus
parvocellular reticular formation
paramedian lobule of cerebellum
pyramidal decussation
reticular formation
superior cerebellar peduncle
substantia gelatinosa
simple lobule
nucleus of the solitary tract
spinal tract of V
supratrigeminal nucleus
superior vestibular nucleus
facial motor nucleus
facial nerve
genu of the facial nerve
hypoglossal motor nucleus
lobules 2 thru 5 of anterior loheof cerebellum
cuneate (Rhoton et al., '66; J acquin et al., '82, '83b; Matesz,
'83; Porter, '86; Pfaller and Arvidsson, '881, vestibular
(Torvik, '56; Matesz, '83; Pfaller and Arvidsson, '881,
trigeminal motor (Kruger et al., '77; J acquin et al., '821, and
facial motor (Clarke and Bowsher, '62) nuclei, as well as the
dorsal motor nucleus of X (J acquin et al., '82, '83b), area
postrema (J acquin et al., '82, '83b), periaqueductal gray
(Arbab et al., ' 88), ventral horn of the cervical spinal cord
(Clarke and Bowsher, '621, and cerebellum (J acquin et al.,
'82, '83b). Most of these "nontrigeminal" projections have
been only superficially described and many have been
disputed, As suggested by J acquin and co-workers ('83b),
some of the controversy in the literature may be due to
interspecies differences and to variability of the methods
used to map the projections, including degeneration, autora-
diography, cobalt labeling, transganglionic transport of
HRP, and electrophysiology.
I n the current study, we investigated the central projec-
tions of the rat trigeminal ganglion to the spinal cord,
contralateral TBNC, and "non-trigeminal" areas of the
CNS by labeling the fibers with the sensitive anterograde
tracer, wheat germ agglutinin-horseradish peroxidase
(WGA-HRP) conjugate. By injecting tracer directly into
each of the three main divisions of the trigeminal ganglion,
we were able t o successfully label the central projections of
the entire trigeminal nerve. I n this manner, the trigeminal
primary afferent projection to the ipsilateral TBNC and
other areas of the brain were demonstrated in exquisite
detail. The main objective of this study was to determine
the extent to which trigeminal primary afferent fibers
convey somatosensory information to areas of the central
nervous system outside of the TBNC. With the exception of
an earlier study by J acquin et al. ('82) who looked at central
projections of the mandibular nerve, this is the first compre-
hensive study dedicated specifically to this issue. This
information should increase our understanding of the
anatomical substrates subserving intermodal convergence
in the central nervous system and the role of brainstem
integrative mechanisms in the higher order discrimination
of intermodal sensory experiences.
METHODS
Twelve young adult, female Sprague-Dawley rats, weigh-
ing 150-220 g, were anesthetized with sodium pentobar-
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE
49 1
bital(25-40 mg/kg body weight, i.p.1 and secured in a head
holder. The left trigeminal ganglion was exposed on the
floor of the middle cranial fossa by removing a small portion
of the left parietal bone and aspirating portions of the
underlying parietal and temporal lobes through a fine-
tipped glass pipette (Marfurt and Turner, 83). Solutions of
WGA-HRP were injected into each of three carefully se-
lected sites along the rostrocaudal length of the ganglion. I n
an effort to label the entire trigeminal ganglion, separate
injections were made into rostral and caudal areas, respec-
tively, of the combined ophthalmomaxillary region, and a
third injection was centered in the mandibular region at the
level of entry of the mandibular nerve. The latter injection
was made into the superficial part of the ganglion in order
to avoid damage to the trigeminal motor root which adheres
to the ventral surface of the ganglion at this location. The
solutions were introduced into the ganglion by using a glass
micropipette (tip diameter, 30-40 pm) connected by a short
length of polyethylene tubing to a 10 pl Hamilton microsy-
ringe mounted on a microinfusion pump. Stability during
the injection procedure was maintained by attaching the
micropipette to the microadvance system of a Kopf small
animal sterotaxic apparatus. In this fashion, the rigidly
mounted pipette tip was lowered under direct visual obser-
vation, using a dissecting microscope at 40 x magnification,
until penetration of the ganglion was made. Small volumes
of 0.2-0.3 ~1 of 2% WGA-HRP in saline were then slowly
infused into the ganglion over a period of 5-6 minutes. The
micropipette was left in place for an additional 10 minutes
after the completion of each injection and thin strips of
gelfoam were applied to the dorsal surface of the ganglion
following removal of the pipette to prevent leakage of tracer
through the dural puncture holes. At the completion of all
three injections, a topical antibiotic was applied to the
wound margins and the scalp incision was closed with
cutaneous sutures.
All animals were killed 4-12 hours after the last injec-
tion. Survival times in excess of 12 hours were tested in
preliminary experiments but provided unsatisfactory re-
sults because they were accompanied by extensive transneu-
ronal labeling of second order neurons at all levels of the
trigeminal brainstem nuclear complex (Marfurt and Ad-
ams, 84) and in adjacent areas of the reticular formation.
Each animal was reanesthetized with pentobarbital and
perfused transcardially with 200 ml of phosphate-buffered
saline (37C, pH 7.3) containing 0.003% heparin and 3%
procaine-HC1, followed by 500 ml of warm fixative consist-
ing of 1% paraformaldehyde-2% glutaraldehyde in 0.1 M
phosphate buffer. The fixation was terminated after 30
minutes by flushing the vasculature with 500 ml of ice-cold
0.1 M phosphate buffer containing 10% sucrose. The brain-
stem, cerebellum, and spinal cord segments C1-T1 were
quickly removed and stored overnight in fresh, ice-cold 30%
phosphate-buffered sucrose. Serial transverse or sagittal
sections were cut at 40 pm on a freezing microtome,
processed as free-floating sections according to the tetra-
methylbenzidine (TMB) procedure of Mesulam (78) and
mounted on chrome alum-gelatin coated slides. Occasional
sections were counterstained with 1% neutral red in 0.1 M
acetate buffer, pH 3.7, in order to distinguish nuclear
boundaries. All other sections were left unstained and were
dehydrated, cleared in xylene, and coverslipped in Per-
mount.
The slides were examined in an Olympus BH-2 light
microscope under brightfield, darkfield, or combined dark-
field-polarized illumination. The locations of WGA-HRP-
labeled fiber processes and their terminal fields were plot-
ted onto a series of line drawings by using a drawing tube
attached to the light microscope. The illustrations included
in this paper depict trigeminal primary afferent projection
patterns for individual rats; however, similar observations,
with some minor differences in innervation density, were
made in all animals.
Relative densities of the terminal fields seen in this study
were graded according to the descriptive terms, heavy,
modest, and sparse. A heavy projection was one in
which the individual grains of reaction product were so
densely packed that they completely occluded the cytoarchi-
tecture of the nucleus in which they were contained through
all levels of focus through the tissue section. Modest
projections contained large numbers of reaction product
granules within a restricted area, but the individual gran-
ules were readily resolved and were usually separated from
one another by varying degrees of clear space (neuropil
devoid of labeled elements). Finally, a sparse projection
was one in which the individual labeled axons and terminals
were widely scattered and/or typically required careful
searching under high levels of magnification to identify
them.
RESULTS
Four to twelve hours after WGA-HRP was injected into
the trigeminal ganglion, terminal labeling was observed
within all four rostrocaudal subdivisions of the ipsilateral
trigeminal brainstem nuclear complex (TBNC), the dorsal
horn of the cervical spinal cord bilaterally, and in numerous
non-trigeminal CNS nuclei (Fig. 1). The anatomical
boundaries of the TBNC, as defined by the distribution of
experimentally labeled afferent nerve fibers, were clearly
demonstrated. Labeled terminals were observed in all dor-
soventral and mediolateral areas of the trigeminal main
sensory nucleus, pars oralis, and pars interpolaris, and in
all laminae of pars caudalis and the contiguous dorsal horn
of the C1 spinal cord (Fig. 2a), suggesting that neurons of
all sizes and in all areas of the trigeminal ganglion had
successfully transported the marker.
Fig. 1 (appears on pages 492495). Line drawings of transverse
sections from the brainstem and upper cervical spinal cord showing the
distribution of reaction product in the ipsilateral TBNC, upper cervical
spinal cord, and surrounding structures following WGA-HRP injections
into the trigeminal ganglion. For the sake of clarity, labeled fibers in the
spinal tract of V have not been illustrated. The stippled shade filmin
levels f-1 indicate areas of extremely high labeling density in the
paratrigeminal nucleus and substantia gelatinosa. I n the periobex
region (level g), the caudal tip of pars interpolaris (inter) separates the
spinal tract of V from the substantia gelatinosa layer (sg) (see J acquin
et al., 88for a detailed anatomical description of this transition zone).
For details concerning trigeminal primary afferent projections to areas
outside of the TBNC, consult text.
492
C.F. MARFURT AND D.M. RAJCHERT
a. MSN
mv
. - - , .
_ _ -
. ..--
._ ,
Figure 1
f. low
inter polar i s
Figure 1 continued
C.F. MARFURT AND D.M. RAJCHERT
. ,
~,
Figure 1 continued
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE 495
dorsal
1.
m.
Figure 1 continued
496
C.F. MARFURT AND D.M. RAJCHERT
Fig. 2. Trigeminal primary afferent projections to the ipsilateral
(a+) and contralateral (d-f) dorsal horn of the upper cervical spinal
cord. Figures a-c illustrate the distribution of reaction product at the
levels of C1, the C2/C3 transition zone, and C5, respectively. d: Fibers
(arrows) crossing the midline at spinal cord segment C1. cc, central
canal. e,f: Terminal labeling in mandibular and ophthalmic regions,
respectively, of the contralateral dorsal horn at C1. a, b, c, X 80; d, e,
f, x 160.
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE 497
In some of the animals, a few faintly labeled neurons
were seen in the mesencephalic nucleus of V, the trigeminal
motor nucleus, and, occasionally, in the reticular formation
adjacent to the dorsomedial region of pars caudalis. The
reaction product in these neurons was completely limited to
the cell body and proximal dendrites.
Projections to the ipsilateral spinal cord
The heavy terminal labeling observed in pars caudalis of
the TBNC (Fig. lh,i) continued uninterupted into the
spinal cord and completely filled the dorsal horn of the C1
segment (Fig. lj, 2a). Labeling intensity was high in all
laminae but was particularly dense in laminae I and 11. I n
C2 and C3, the amount of terminal labeling decreased and
was concentrated mainly in a wedge-shaped, intermediate
area of the dorsal horn, as well as in laminae I throughout
its entire mediolateral extent (Figs. lk,l, 2b). Only modest
amounts of reaction product were observed in levels C4 and
C5, where most of it was concentrated in laminae I, 11, and
V (Figs. lm, 2c). A few isolated fibers descended as far
caudally as C6 and C7; the latter axons ended almost
exclusively in lamina I.
Contralateral projections
Numerous fibers crossed the midline at the level of caudal
pars caudalis and the upper cervical spinal cord and termi-
nated on the contralateral side (Fig. 3). The majority of the
fibers crossed over between the midlevel of the pyramidal
decussation and the caudal border of C2; however, occa-
sional fibers also decussated within rostral caudalis and in
C3. All of the fibers crossed the midline in the dorsal gray
commissure posterior to the central canal (Fig. 2d). After
reaching the opposite side, some of the fibers ascended or
descended for several millimeters before terminating and,
as a result, reaction product was observed on the contralat-
eral side as far rostral as the obex and as far caudal as the
C5 spinal cord.
Most of the crossed fibers originated from dorsomedial
(mandibular) and ventrolateral (ophthalmic) regions of
ipsilateral pars caudalis and the cervical dorsal horn (Fig.
li-1) and projected to corresponding areas of the contralat-
eral side (Figs. 2e,f, 3). I n contrast, connections between
ipsilateral and contralateral intermediate (maxillary) re-
gions of the two sides were relatively sparse. Some interani-
mal variability was noted as regards the intensity of the
divisional crossed representations, i.e., in some animals, the
number of crossed mandibular fibers exceeded the number
of crossed ophthalmic fibers, (as in the animal illustrated in
Fig. 3), whereas, in other animals the reverse was true. At
levels of maximum crossed projection (i.e., caudal pars
caudalis and Cl), terminal labeling extended into all lami-
nae of the contralateral side (Figs. 2e,f, and 3, levels b-d),
although it was most heavily concentrated in laminae 111-V
and in the deeper part of lamina 11. By comparison, more
rostral levels of caudalis (Fig. 3, level a) and more caudal
segments of the cord (Fig. 3, levels +g), contained only
sparse amounts of contralateral labeling and it was con-
fined largely to laminae I, 11, and V.
Examination of material sectioned in the sagittal plane
provided further details of the crossed projection (Fig. 4).
The decussating mandibular and ophthalmic fibers crossed
the midline at regular intervals as a collection of 30-40
evenly spaced nerve fascicles. Each fascicle contained approx-
imately 5-40 labeled fibers and was separated from its
rostral and caudal neighbors by a distance of approximately
250-400 bm.
No contralateral projections were observed at levels of
the TBNC rostral to the obex (i.e., pars interpolaris, pars
oralis, or the main sensory nucleus).
Projections to non-trigeminal areas of the
brainstemandtothecerebellum
Terminal labeling was also observed in a variety of
non-trigeminal areas of the central nervous system.
Perhaps the densest labeling seen in this study was in the
ipsilateral paratrigeminal nucleus (Figs. lf, 5a). The latter
nucleus consisted of multiple, irregular-shaped islands of
neuropil embedded in the dorsal half of the spinal tract of V
from slightly caudal to the obex to the midlevel of pars
interpolaris. The density of the terminal labeling in this
area was so great (exceeding even that of the adjacent
TBNC) that the cytoarchitecture of the underlying neuropil
was totally obscured.
Trigeminal primary afferent fibers also projected very
heavily to the ipsilateral nucleus of the solitary tract (Figs.
lc-h, 5). The density of the projection varied considerably
along the rostrocaudal axis of the nucleus, being heaviest in
the intermediate region (at the level of rostral and middle
pars interpolaris), moderate to light rostrally (adjacent to
pars oralis) and sparce caudally (periobex region). The
terminal labeling was confined largely to the lateral part of
the nucleus at most levels (Fig. 5b); however, in the area of
the greatest innervation density (adjacent to pars interpo-
laris), heavy labeling also extended into the medial area of
the nucleus (Figs. le,f, 5a). Most of the labeled fibers
entered the nucleus from the dorsal part of the spinal
nucleus of V, suggesting a largely mandibular origin for
these fibers. In the periobex region, a small amount of
reaction product was observed in the midline commissural
nucleus of X (Fig. l h); occasional fibers also crossed at this
level to terminate on the opposite side (Fig. lg).
Trigeminal primary afferent fibers also projected in signif-
icant numbers to the supratrigeminal nucleus (supra V)
(Figs. l a, 6a), and a few fibers projected to the dorsolateral
region of the trigeminal motor nucleus (Figs. l a, 6a,b). The
terminal labeling in supra V was heaviest laterally (i.e.,
adjacent to the dorsomedial part of the trigeminal main
sensory nucleus) and relatively light medially; occasional
fibers were observed among the cells of the mesencephalic
nucleus of V (Fig. la). The scattered fibers that entered the
motor nucleus of V (Fig. 6b) originated from the dorsome-
dial region of the neighboring main sensory nucleus, suggest-
ing a mandibular origin for these fibers.
Numerous fibers terminated in the reticular formation
adjacent to pars oralis and pars interpolaris (Figs. lb-f, 6c,
7) and in lamina V of pars caudalis2 (Figs. lg-i, 6d). In
contrast, labeled terminals were not observed in the reticu-
lar formation adjacent to the main sensory nucleus (Fig.
la), except for some minor labeling near its caudal border
with pars oralis. At all rostrocaudal levels, most of the
labeling was concentrated in areas of the reticular forma-
tion adjacent to dorsal (mandibular) areas of the TBNC,
whereas the reticular formation adjacent to intermediate
(maxillary) and ventral (ophthalmic) regions of the TBNC
contained progressively smaller amounts of label. The
*The area ventromedial to the magnocellular layer (lamina IV) of pars
caudalis was regarded, up until the late 1960s, as a part of the medullary
reticular formation. However, the results of numerous recent anatomical
and physiological investigations have shown that this region (which is in
many ways comparable to lamina V of the spinal cord dorsal horn) is an
integral part of pars caudalis.
498
C.F. MARFURT AND D.M. RAJCHERT
m
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE
-
499
ipsiiaterai contral ateral
Fig. 4. Line drawing of the trigeminal primary afferent projection to
contralateral pars caudalis and the dorsal horn of the upper cervical
spinal cord as seen in sagittally sectioned material. The area shown in
the drawings at the right is enclosed by the box in the small (top)
orientation figure at left. The letters a-e in the bottom orientation
figure identify the relative mediolateral position of each parasagittal
section. The dashed lines in the figures at the right indicate the
heaviest RF labeling seen in this study was observed in the
dorsal RF adjacent to pars interpolaris and subjacent to the
nucleus of the solitary tract (Figs. le,f; 6c; 7).
Fig. 3. Line drawings showing the distribution of trigeminal pri-
mary afferent fibers to contralateral pars caudalis (a,b) and the dorsal
horn of C1 (c,d), C2 (e,n, and C3 (g). The numbers to the right of each
section indicate the distance in millimeters caudal to the obex.
approximate limits of pars caudalis, C1, and C2 (see labels in level b).
Mandibular and ophthalmic nerve fibers crossed the midline from the
pyramidal decussation to C2 in a series of evenly spaced, multiple nerve
fascicles (level a). When the fibers reached the contralateral dorsal
horn, the preterminal axons ascended and descended to intermingle
with fibers arriving from adjacent nerve fascicles to form longitudinal
columns of terminals.
Modest amounts of very fine terminal labeling were also
observed in the lateral part of the cuneate nucleus (Figs. lg,
8, 10a) and a few fibers were seen in the vestibular nuclei
(Figs. l b, 9, 10b,c). The trigeminovestibular fibers exited
the dorsomedial region of the TBNC near the junction of
the main sensory nucleus and pars oralis and terminated
mainly in the lateral and superior vestibular nuclei.
Finally, still other trigeminal primary afferent fibers
(approximately 15-25 per animal) terminated in the ipsilat-
500
C.F. MARFURT AND D.M. RAJCHERT
Fig. 5. a: Photomontage showing heavy terminal labeling in the
paratrigeminal (para V) nucleus at the level of middle pars interpolaris.
The dark appearance of the para V nucleus in this black and white
micrograph is an optical phenomenon caused by excessive superimposi-
tion of reaction product granules interfering with the normal scattering
of light. Heavy labeling is also seen at this level in the medial and lateral
regions of the nucleus of the solitary tract (sol). b Labeling in the
nucleus of the solitary tract at the level of caudal pars oralis. Reaction
product is concentrated in the lateral part of the nucleus (arrow). tr sol,
tractus solitarius. c: Terminal labeling in the nucleus of the solitary
tract (sol) and subjacent parvocellular reticular formation (pcRF) as
viewed in the sagittal plane. Dorsal is towards the top and rostra1 is to
the right. a, b, c, x 80.
era1 cerebellum (Fig. 11). The latter fibers exited the
dorsomedial pole of the TBNC at the level of the main
sensory nucleus, (Fig. 10d) entered the cerebellum via the
superior cerebellar peduncle (Fig. lOe), and projected to
widespread areas of the ipsilateral cerebellum, including
crura I and I1 of the ansiform lobule, the pyramidal lobule,
the paraflocculus, lobules 4 and 5 of the anterior lobe, and
the interposed and lateral (dentate) cerebellar nuclei (Fig.
l0n. Overall, slightly more fibers entered the posterior lobe
than the anterior lobe (Fig. 11). Although the axons were
well labeled in these experiments, terminal labeling in the
cerebellum was not observed.
The main findings of the present investigation are sum-
marized in Figure 12 and in Table 1.
DISCUSSION
projections to the ipsilateral dorsal horn of the cervical
spinal cord from C1 to C7, contralateral pars caudalis and
dorsal horn of Cl-C5, the nucleus of the solitary tract, the
paratrigeminal, supratrigeminal, cuneate, vestibular, and
motor V nuclei, the reticular formation, and the cerebel-
lum. These results suggest that the transmission of pri-
mary somatosensory information from the head and face is
perhaps more complex than previously recognized for this
system. Many of the projections demonstrated in this study
(e.g., to the paratrigeminal and supratrigeminal nuclei, the
nucleus of the solitary tract, and reticular formation)
appear quantitatively significant and, as a result, may be of
considerable functional importance, whereas other projec-
tions (e.g., to the vestibular, motor V and cuneate nuclei,
the C3-C7 spinal cord, and cerebellum) are relatively minor
and their physiological significance is perhaps limited.
Technical collsiderations
The WGA-HRP labeling procedure used in the present
study produced excellent labeling of the trigeminal primary
afferent projection. Certain aspects of the results (e.g., the
The results of this study have demonstrated that rat
trigeminal primary afferent neurons project not only to the
ipsilateral TBNC, but also to other areas of the brain.
Injections of the sensitive neuroanatomical tracer, WGA-
HRP, into the trigeminal ganglion produced well-labeled
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE
50 1
Fig. 6. a: Terminal labeling in the supratrigeminal nucleus (supra
V) and trigeminal motor nucleus (mV). b: Higher magnification of the
area illustrated in a, showing scattered labeled nerve fibers (arrows)
in the dorsolateral region of the trigeminal motor nucleus. c: Labeling
in the parvocellular reticular formation (pcRF) at the level of pars
interpolaris d Dense terminal labeling in the ventromedial (lamina V)
region of pars caudalis. The transversely oriented fibers at lower right
(arrows) are crossing to the contralateral side. a, c, X 80; b, d, X 160.
presence of strong labeling throughout the dorsoventral
extent of the TBNC and in all laminae of pars caudalis)
strongly suggest that the majority of cells in all regions of
the ganglion, and representing the full range of cell diame-
ters, participated in the transport process. The procedure
generated a clear and precise picture of trigeminal primary
afferent connectivity that provides more robust staining of
the primary afferent projection than does autoradiography
(Kruger et al., 771, avoids the problems of inconsistant
staining of smaller fibers and frequent staining artifacts
that often accompany silver degeneration experiments,
and, unlike cobalt labeling procedures, allows satisfactory
filling of both large and small diameter fibers (Matesz, 83).
Several projections that were unlabeled by cobalt filling
techniques (e.g., the substantia gelatinosa layer of pars
caudalis, the paratrigeminal nucleus, and the supratrigemi-
nal nucleus) were strongly labeled in this study.
Survival times in excess of 8-12 hours are accompanied
by strong transneuronal transport of WGA-HRP to second
order neurons of the TBNC (Marfurt and Adams, 84;
Pfaller and Arvidsson, 88); therefore, only very short survi-
val times were used in this study. In preliminary experi-
ments employing straight, unconjugated HRP (unpub-
lished observations), transneuronal transport of tracer into
second order cells was not observed; however, overall label-
ing intensity was reduced and some of the minor projections
(e.g., to the vestibular nuclei and to the cerebellum) were
poorly labeled or absent.
In most of the animals examined in this study, weob-
served a small number (10-20) of very faintly labeled
neurons in the motor V and mesencephalic V nuclei and in
the reticular formation adjacent to the dorsomedial tip of
the MSNJ pars oralis transition zone. J acquin and co-
workers (83a) have reported that the latter cells belong to
the inferior salivatory nucleus and that their axons course
through the mandibular region of the trigeminal ganglion
to reach the otic ganglion. WGA-HRP injected into sensory
ganglia is not usually taken up by (motor) axons of passage
(Pugh and Kalia, 82); thus, the retrogradely labeled cells
seen here probably resulted from mechanical or pressure-
induced injury to the motor axons by the pipette tip. The
reaction product in mes V was confined to the cell somata
and the labeling in motor V and the inferior salivatory
nucleus was restricted to the cell somata and proximal
dendrites, and, therefore, did not interfere with the interpre-
tation of the data.
502
C.F. MARFURT AND D.M. RAJCHERT
anatomical substrate for certain types of craniofacial pain,
for example, atypical neuralgia (Kerr, '721, and the fact that
much of the labeling observed in the cervical dorsal horn in
the present study was found in laminae I, 11, and V may
indirectly support this hypothesis.
Primary trigeminospinal projections do not constitute
the sole source of trigeminal input to the spinal cord, for
there are also significant trigeminospinal projections from
second order neurons in the spinal trigeminal nucleus
(Ruggiero et al., '81). The latter fibers have been shown to
project most heavily to cervical levels of the cord; however,
other fibers descend as far caudal as the lumbosacral level
(Ruggiero et al., '81). The physiological significance of the
dual (primary and secondary) trigeminospinal pathways is
uncertain; however, they may cooperate in certain trigemi-
nospinal reflexes, including the diving, oculocardiac, and
nasocardiorespiratory reflexes (Miles, '79).
Contralateral projections
The results of this study have confirmed previous reports
of a projection from the trigeminal ganglion to the contralat-
era1 caudal level of pars caudalis and dorsal horn of the
upper cervical spinal cord. The intensity of the projection
revealed here appears to be slightly greater than that
shown by most earlier workers (Torvik, '56; Clarke and
Bowsher, '62; Matesz, '83; Pfaller and Arvidsson, '88)
suggesting a more complete labeling of this pathway by the
sensitive WGA-HRP anterograde tracing procedure.
Although the WGA-HRP bulk labeling technique pro-
vides little information concerning the peripheral distribu-
tion of the fibers that contribute to this projection, the
pronounced distribution of these fibers to dorsomedial and
kentrolateral areas of the nucleus suggests mainly mandib-
ular and ophthalmic The results of most previous
investigations have suggested that the mandibular division
is the major, if not only, source of crossed trigeminal
primary afferent projections; therefore, the finding in this
study of an extremely robust ophthalmic contribution is of
particular interest. Similar observations have recently been
reported by J acquin and co-workers ('90) using bulk HRP
Fig. 7. Terminal labeling (white arrows) in the reticular formation
medial to caudal pars interpolaris and ventral to the nucleus of the
solitary tract (sol). x 80.
hjections to the ipbilateral
cer vi cal sp~cor d
The results of this study have confirmed and extended
the reports of other workers that trigeminal primary
afferent neurons project heavily to the dorsal horn of the C1
and C2 spinal cord, and in lesser amounts to the cervical
dorsal horn from C3-C7 (J acquin et al., '83; Pfaller and
Arvidsson, '88). Previously, it had been suggested that
these fibers descended within the cord only as far caudally
as C2 or C3 (Torvik, '56; Clarke and Bowsher, '62; Kerr,
'63; Marfurt, '81; Panneton and Burton, '81; Shigenaga,
'86a1, and it is probable that the true magnitude of this
projection was underestimated due to the lower sensitivi-
ties of the degeneration and transganglionic HRP transport
procedures used in the earlier studies.
The dorsal horns of C1 and C2 also receive extensive
primary afferent input from dorsal root ganglia C2 and C3,
and it is probable that some fibers from the trigeminal and
cervical dorsal root ganglia converge on common second
order neurons (Kerr, '72; Sessle et al., '86; Pfaller and
Arvidsson, '88). This convergence may represent a mecha-
nism by which the brain "fuses" the central representa-
tions of peripheral structures located in the transition zone
between the innervation territories of the trigeminal and
C2 cervical nerves (e.g., the caudal scalp, retroauricular
area, and peripheral face) to form a continuous somatotopi-
cal map. Convergence of trigeminal and spinal afferents on
common neurons within the dorsal horn of the upper
cervical spinal cord has also been suggested as a possible
transport techniques.
The results of several tranganglionic HRP tracing experi-
ments suggest that most of the decussating fibers originate
from trigeminal nerve branches with peripheral receptive
fields extending to, or beyond, the midline, including the
inferior alveolar, mental, and lingual nerves (Arvidsson and
Gobel, '81; Marfurt, '81; J acquin et al., '83b; Hamilton
and Norgren, '84; Takemura et al., '87). Furthermore, it
has been suggested that midline cutaneous, non-nocicep-
tive fibers may contribute most heavily to this decussation
(J acquin et al., '90). In contrast, nerves that distribute only
to the ipsilateral side of the face, e.g., the auriculotemporal
and mylohyoid nerves (J acquin et al., '83b), and nerves to
the cornea (Marfurt and DelToro, '87; Marfurt and Echten-
kamp, ,881, do not contribute to the crossed projection in
the brainstem. An analogous situation exists within the
spinal cord where crossed primary afferent projections are
abundant in spinal cord segments that receive input from
midline areas such as the neck, trunk, and tail but are
largely lacking in spinal cord segments that innervate the
Fig. 8. Terminal labeling (arrows) at three different levels of the
cuneate nucleus. Reaction product is concentrated in the extreme
lateral area of the nucleus and adjacent to the dorsomedial edge of the
spinal tract of V. Numbers to the right of each figure indicate the
distance in millimeters caudal to the obex.
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE 503
Figure 8
504
C.F. MARFURT AND D.M. RAJCHERT
+3.25
Fig. 9. Labeled fibers in the ipsilateral vestibular complex. Numbers to the right of each figure indicate
the distance in millimeters rostral to the obex.
extremities (Culberson et al., '79; Light and Perl, '79;
Pfaller and Arvidsson, '88; Rethelyi et al., '79). Thus,
bilateral projections of sensory information from intraoral
and facial tissues near the midline may be important to the
central fusion of sensory maps of the right and left sides of
the head (Smith, '86) and to the rapid, bilateral activation
of brainstem circuitry subserving withdrawal or protective
reflexes (J acquin et al., '90).
Crossed trigeminal primary afferent projections to more
rostral levels of the TBNC (main sensory nucleus, pars
oralis, and pars interpolaris) were not observed in this
study, nor have they been observed by other workers using
either cobalt labeling (Matesz, '83) or HRP transport
(J acquin et al., '83b, '90; Pfaller and Arvidsson, '88) tech-
niques. Crossed projections at rostral levels have been
reported using degeneration techniques (Torvik, '56; Clarke
and Bowsher, '62; Westrum et al., '76); however, there has
been some suggestion that the crossed degeneration rag-
ments observed in the latter studies resulted from trans-
neuronal degeneration of second order neurons (Gobel and
Binck, '77).
Projections to brainstem nuclei
Paratrigeminal nucleus. The results of this study
have demonstrated an extremely dense trigeminal primary
afferent projection to the paratrigeminal nucleus (J acquin
et al., '88). The precise role of the paratrigeminal nucleus in
somatosensory processing remains unclear; however, re-
cent studies of its afferent and efferent connectivity have
provided clues to its possible functions. The nucleus re-
ceives afferent input from pharyngeal and soft palatal
branches of IX and X (Altschuler et al., '89) and from
intraoral branches of the trigeminal (Shigenaga et al.,
'86a,b; Takemura et al., '87; Nazruddin et al., '89) and
sends efferent axons to the parabrachial (Cechetto et al.,
'85; Panneton and Burton, '85; Menetrey et al., '871,
solitary, and reticular formation nuclei (Menetrey et al.,
'87). On the basis of its known connectivity, and by virtue of
the fact that the nucleus increases its metabolic activity
during hibernation in ground squirrels (Kilduff et al., '83),
it has been postulated that para V may function in the
regulation of autonomic activity, perhaps by monitoring
perioral temperature (Cechetto et al., '85; Menetrey et al.,
'87). Alternatively, the abundance of substance P and
enkephalinergic inputs to this nucleus (reviewed by Phelan
and Falls, '89) suggests a role for para V in pain processing.
The demonstration of a dense trigeminal primary afferent
input to the paratrigeminal nucleus in this study does not
contradict either of these hypotheses.
The massive trigemino-
solitary projection seen here confirms previous descriptions
of this projection in multiple animal species as studied by
Nucleus of the solitary tract.
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE
505
Fig. 10. a: Labeled fibers (arrow) and terminals (arrowheads) in the
lateral region of the main cuneate nucleus (cun). b,c: Labeled fibers
(arrows) in the vestibular nuclei. d Labeled trigeminocerebellar axons
(arrows) entering the superior cerebellar peduncle. e,f: Labeled axons
(arrows) in the superior cerebellar peduncle ( e, parasagittal section)
and interposed nucleus of the cerebellum (0. a, b, d, f, X 80; c, e, X 160.
degeneration (Torvik, '56; Clarke and Bowsher, '62; Ken-,
'63; Rhoton et al., '66) autoradiography (Kruger et al . , '77;
Beckstead and Norgren, '79; Contreras et al., '821, and HRP
transport techniques (e.g., J acquin et al., '82; Nomura et
al., '84; Pfaller and Arvidsson, '88). There is general agree-
ment among authors that the fibers 1) project with varying
intensity to all rostrocaudal levels (gustatory and nongusta-
tory) of the nucleus of the solitary tract; 2) terminate most
heavily in lateral and ventrolateral areas of the nucleus,
although many fibers also enter the medial area at levels of
506
C.F. MARFURT AND D.M. RAJCHERT
4
a
J
K
I-
v)
$
cu
I
3
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE 507
SUMMARY DIAGRAM OF TRIGEMINAL
PRIMARY AFFERENT PROJECTIONS
I
Q Trigeminal
Ganglion
/---
Fig. 12. Summary diagram illustrating the main findings of the
present study. Trigeminal primary afferent projections to the ipsilateral
cervical spinal cord, contralateral pars caudalis and spinal cord, the
cerebellum, and to numerous non-trigeminal brainstem nuclei are
illustrated. For the sake of clarity, the robust trigeminal primary
afferent projection to the ipsilateral TBNC and to the dorsal horn of C1
and C2 are not shown.
middle and rostral pars interpolaris; and 3) are primarily of
mandibular origin. The results of HRP transganglionic
transport experiments have shown that the heaviest projec-
tions derive from neurons that innervate tissues of the oral
cavity via the lingual (Whitehead and Frank, 83; J acquin et
al., 83b; Nomura et al., 84; Hamilton and Norgren, 84;
Shigenaga et al., 86b; Takemura et al., 871, inferior
alveolar (J acquin et al., 83b; Takemura et al., 87), buccal
Fig. 11. Distribution of labeled axons (arrows) in the ipsilateral
cerebellum as seen in sagittally sectioned material. Numbers to the left
of each figure indicate the distance in millimeters lateral to the midline.
(Takemura et al., 871, and palatal (Nomura et al., 84;
Arvidsson and Hellstrand, 881 nerves; however, limited
projections from the mylohyoid and auriculotemporal nerves
have also been reported (J acquin et al., 83b). In addition, a
trigeminosolitary projection from the ophthalmic division
of the trigeminal ganglion has been shown using autoradio-
graphic labeling procedures in monkeys (Beckstead and
Norgren, 79).
Functionally, the nucleus of the solitary tract is orga-
nized into a rostral, gustatory region, and a caudal region
that is viscerotopically organized to receive afferent input
from respiratory, cardiovascular, and gastrointestinal
sources. Trigeminal primary afferents project to all rostro-
caudal levels of the nucleus of the solitary tract, suggesting
roles for these fibers in all of these functions. For example,
trigeminal inputs to caudal, nongustatory levels of the
nucleus of the solitary tract may be important in the
integration of sensory information from the oral cavity,
pharynx, and esophagus during mastication and deglutition
and in the generation of certain somatovisceral reflexes.
Intermodal convergence of taste and somesthetic inputs
from the tongue and surrounding tissues of the oral cavity
in the gustatory region of the nucleus of the solitary tract
may provide a mechanism whereby the texture, hardness,
temperature, and other characteristics of food substances
may impart subtle overtones to ones perception of the
flavor of particular foods. Thus, chilling or warming a
drink may alter ones perception of its taste, and capsaicin
(a pungent compound found in hot peppers that activates
polymodal nociceptors) contributes to the hotness or
spice associated with certain exotic dishes. Multimodal
lingual units (neurons that respond to both taste and tactile
or thermal stimuli) have been found by electrophysiological
investigations in the nucleus of the solitary tract (Makous
et al., 63; Halpern and Nelson, 651, as well as at higher
levels of the gustatory system, i.e., the thalamus and
cerebral cortex (Yamamoto et al., 81).
The dense projection to the supra-
trigeminal nucleus seen in this study confirms previous
findings of other workers based on WGA-HRP (Pfaller and
Arvidsson, 88) and tritiated amino acid (Contreras et al.,
82) injections into the trigeminal ganglion, as well as
transganglionic transport of WGA-HRP along peripheral
branches of the trigeminal nerve (J acquin et al., 82, 83b;
Hamilton and Norgren, 84; Shigenaga et al., 88; Take-
mura et al., 87). Some of the HRP tracing studies were
accompanied by heavy labeling in the mesencephalic nu-
cleus of V (Takemura et al., 87; Pfaller and Arvidsson, 88;
Shigenaga et al., 88), and since central axons of mes V
neurons also project heavily to the supratrigeminal nucleus
(Matesz, Sl), there was some question concerning the
origin of the supra V labeling. However, the results of the
current study, and of an earlier transganglionic HRP
transport study in which the tracer was applied selectively
to the sensory mandibular nerve fibers while excluding the
motor axons (J acquin et al., 83b), clearly reveal a trigemi-
nal ganglion origin for this labeling. The projections to
supra V from the trigeminal ganglion arise primarily from
neurons that distribute through intraoral branches of the
mandibular nerve, including the lingual, inferior alveolar,
superior alveolar, and buccal nerves (J acquin et al., 83b;
Hamilton and Norgren, 84; Takemura et al., 87) .
Additional projections to supra V from extraoral nerves,
e.g., the auriculotemporal and mylohyoid, have been
reported by some (J acquin et al., 83b) but not all (Take-
mura et al., 87) workers. Whether or not mes V and
Supratrigeminal.
508
C.F. MARFURT AND D.M. RAJCHERT
TABLE 1. Intensity of Trigeminal Primary Afferent Projections to Various CNS Nuclei
Modest Sparse Absent or unconfirmed
Heavy
para V pcRF Vestibular Facial motor nucleus
Nucleus of the solitary tract
Supra V Contralateral caudalis and dorsal horn of C1-C2 Cerebellum Dorsal motor nucleus of X
Ipsilateral dorsal born of C3 and C4 MotorV
Cuneate Areapostrema
Ipsilateral dorsal horn of C5-C7
Contralateral dorsal horn of C3-C5
Ventral horn of cervical spinal cord
Periaqueductal gray
trigeminal ganglion cells project to similar, or different,
populations of supra V cells is unknown.
The supratrigeminal nucleus also receives afferent input
from the contralateral supratrigeminal nucleus and the
medullary reticular formation bilaterally (Rokx et al., '86),
and cells of this nucleus project bilaterally to the trigeminal
motor nuclei and medullary reticular formation, and ipsilat-
erally to the facial and hypoglossal motor nuclei (Rokx et
al., '86). Electrophysiological investigations have shown
that the Supra V nucleus contains interneurons that are
inhibitory to trigeminal motor neurons and are involved in
the jaw opening reflex (J erge, '63). Thus, the sum total of
available evidence suggests that the supra V nucleus is an
important center for the control and integration of oral
motor behavior.
The results of this study have also demon-
strated a direct trigeminal primary afferent projection to
the motor nucleus of V. The existence of such a projection
has been much debated in the literature, having first been
described in normal material (Ram6n y Cajal, '09; Astrom,
'53), and subsequently refuted on the basis of negative data
generated from experimental degeneration (Szentagothai,
'48; Torvik, '561, and cobalt labeling (Matesz, '83) tech-
niques. More recently, however, Kruger and co-workers
('77) and J acquin et al. ('83b) have confirmed this projec-
tion using autoradiography and bulk HRP transport proce-
dures, respectively. Most of the HRP-labeled fibers seen in
the current study were located in the dorsolateral part of
the motor V nucleus, which, according to the myotopic
organization of the rat motor V nucleus (Limwongse and
DeSantis, '77; Mizuno et al., '75; Lynch, '85), contains cells
that innervate the jaw closing muscles (masseter, tempo-
ralis, and medial pterygoid).
Functionally, the scant trigeminal primary afferent input
to motor V seen in this study must be viewed in the context
of additional, more substantial inputs to the motor V
nucleus from the supratrigeminal, intertrigeminal and
mesencephalic V nuclei, the dorsomedial region of the main
sensory nucleus, and various areas of the reticular forma-
tion (Vornov and Sutin, '83). Although their physiological
role may be limited, it is tempting to speculate that
trigeminal primary afferent fibers to the motor V nucleus
are activated by contact with food in the oral cavity during
mastication or by stimulation of periodontal afferents and
that these inputs assist in the control of masticatory
movements. Alternatively, painful orofacial stimuli in exper-
imental animals induces vigorous jaw opening and tongue
retraction, suggesting a reflex mechanism for protection of
intraoral structures from injury during chewing (Brat-
zlavsky, '77).
The results of this study con-
firm a significant trigeminal primary afferent projection to
the reticular formation bordering the spinal nucleus of V,
and that the region of maximum input is to the area
dorsomedial to the caudal one half of pars interpolaris
(Torvik, '56; Clarke and Bowsher, '62; Kruger et al., '77;
Motor V.
Reticular formation.
J acquin et al., '82, '83b; Matesz, '83; Pfaller and Arvidsson,
'88). Studies based on the transganglionic transport of HRP
suggest that much of the input to this area is derived from
branches of the mandibular nerve, including the inferior
and superior alveolar nerves, and the lingual (J acquin et al.,
'83b; Hamilton and Norgren, '84; Shigenaga et al., '86b;
Takemura et al., '87). Some of this information is probably
transmitted into the reticular activating system or to
cranial nerve motor nuclei for reflex purposes.
Caudal to the obex, numerous labeled fibers extend into
the area ventromedial to the magnocellular layer of pars
caudalis. This region, once considered as a part of the
medullary reticular formation, is currently regarded by
most workers as an integral component of nucleus caudalis
with particular importance in the mediation of orofacial
pain (Nord and Kyler, '68; Nord and Ross, '73; Tiwari and
King, '74) and it has been redesignated lamina V of the
"dorsal horn of the medulla" (Gobel et al., '77).
The modest projection to the cu-
neate nucleus seen in this study confirms reports of a
trigeminal primary afferent input to this nucleus by other
workers (Rhoton et al., '66; J acquin et al., '82, '83b; Matesz,
'83; Porter, '86; Pfaller and Arvidsson, ' 88) . The peripheral
distributions of the trigeminal cells that project to the
cuneate remain largely unknown; however, the results of
two recent transganglionic HRP transport studies suggest
that some of these fibers in rats derive from the inferior
alveolar nerve (J acquin et al., '83b1, whereas others in
monkeys innervate the extraocular muscles (Porter, '86).
The ventrolateral location of the cuneate terminal labeling
observed in the present study mimics the distribution of
trigeminal primary afferent labeling in the monkey cuneate
nucleus (Rhoton et al., '66; Porter, '86). In cats, the
ventrolateral region of the cuneate nucleus receives promi-
nent inputs from muscle and cutaneous tissues of the neck
(Abrahams et al., '79, '84). Thus, the ventrolateral area of
the cuneate nucleus may be important for the integration of
different types of sensory information (from neck muscles
and cutaneous afferents, as well as from the head and face)
necessary for the coordination of head, neck, and eye
movements (Porter, '86).
The results of this study have demon-
strated a scant projection from the trigeminal ganglion to
the ipsilateral superior and lateral vestibular nuclei, con-
firming similar observations by a handful of other workers
(Torvik, '56; Matesz, '83; Pfaller and Arvidsson, '88). The
vestibular nuclei also receive substantial inputs from spinal
cord levels. The latter fibers carry mainly proprioceptive
information from cervical levels (in particular the central
cervical nucleus) and terminate largely in the medial and
descending (inferior) vestibular nuclei (Neuhuber and Zen-
ker, '89; McKelvey-Briggs et al., '89), where they overlap
extensively with afferent inputs from the vestibular nerve
(Boyle and Pompeiano, '81). Although the limited nature of
the trigeminovestibular projection seen here would argue
against a significant physiological role, their predominantly
Cuneate nucleus.
Vestibular.
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE
509
mandibular origin makes it tempting to speculate that
these inputs may influence in a minor way vestibular
nuclear control of head posture and gaze stabilization
during normal mastication.
Cerebellar projections. Multiple routes exist for the
transmission of trigeminal sensory information to the
cerebellum, including secondary trigeminocerebellar projec-
tions from the TBNC (especially, the MSN and pars interpo-
laris, Steindler, 77; Watson and Switzer, 78; Ikeda, 79;
Matsushita et al., 82; Mantle-St. J ohn and Tracey, 87) and
the trigemino-olivary-cerebellar pathway (Huerta et al.,
83). The results of this study show that in rats there is an
additional, albeit minor, pathway by which trigeminal
sensory information is relayed directly to the cerebellum
without necessity of brainstem relays. Although the exist-
ence of primary trigeminocerebellar fibers had been previ-
ously denied on the basis of degeneration (Torvik, 56) and
cobalt labeling (Matesz, 83) techniques, they have recently
been shown by applying HRP to the whole mandibular
nerve (J acquin et al., 82, 83b). The peripheral target
tissues supplied by these afferents, and the nature of the
afferent information being relayed centrally, remains spec-
ulative; however, J acquin and co-workers (83b) have shown
through the use of HRP transganglionic transport that at
least some primary trigeminocerebellar afferents distribute
through the inferior alveolar and lingual nerves. Primary
and secondary trigeminocerebellar afferents enter the ipsi-
lateral cerebellum separately (the former through the
superior cerebellar peduncle and the latter via the inferior
cerebellar peduncle); however, once within the cerebellum
they appear to converge on similar cerebellar folia and deep
nuclei, including cms I and 11, the paramedian lobule,
lobulus simplex, lobules 3 and 4 of the anterior lobe, and the
interposed and lateral deep cerebellar nuclei (Watson and
Switzer, 78; Somana et al., 80; J acquin et al., 82).
Trigeminal tactile projections to many of these areas have
also been demonstrated electrophysiologically (Shambes et
al., 78). The limited nature of the scant primary trigemino-
cerebellar projection shown here suggests that its physiolog-
ical significance (when contrasted with the potential wealth
of information carried through the secondary trigeminocer-
ebellar and trigemino-olivary-cerebellar pathways) may be
minimal.
Unconfirmed projections. Finally, some comment
remains to be made concerning certain negative findings
of the current study. For example, the present study
provided no evidence for a trigeminal primary afferent
projection to the VII motor nucleus (Clarke and Bowsher,
62). Others have shown, however, that the rat VII motor
nucleus receives significant trigeminofacial input via sec-
ond order neurons in the adjacent TBNC (Erzurumlu and
Killackey, 79). Secondary trigeminofacial projections may
relay vibrissal sensory information to facial motor neurons
that innervate the vibrissal musculature (Watson and
Switzer, 78), thereby influencing whisking behavior.
Nor did the present study provide any evidence for direct
trigeminal afferent input to the ventral horn of the cervical
spinal cord (Clarke and Bowsher, 62), periaqueductal gray
(Arbab et al., 88), area postrema (J acquin et al., 821, or the
dorsal motor nucleus of X (J acquin et al., 82). Careful
analysis of the micrographs provided by J acquin and co-
workers (82) have led Norgren and Hamilton (84) to
conclude that the labeled terminals ascribed to dorsal X in
J acquins work are, in reality, situated in the reticular
formation immediately lateral to the nucleus of the solitary
tract.
ACKNO-MENTS
The authors are indebted to Christine Coleman, Larry
Ellis, and Mark J ones for providing excellent technical
assistance, and Dr. J an Arvidsson for numerous valuable
comments on an earlier version of this manuscript. This
work was supported by a grant from the National Institute
of Health, EY05717 (C.F.M.)
Abrahams, V.C., G. Anstee, F.J. Richmond, and P.K. Rose (1979) Neck
muscle and trigeminal input to the upper cervical cord and lower medulla
of the cat. Can. J. Physiol. Pharmacol. 57:642-651.
Abrahams, V.C., F.J. Richmond, and J. Keane (1984) Projections fromC2
and C3 nerves supplying muscles and skin of the cat neck: A study using
transganglionic transport of horseradish peroxidase. J. Comp. Neurol.
230:142-154.
Altschuler, S.M., X. Bao, D. Bieger, D. Hopkins, and R. Miselis (1989)
Viscerotopic representation of the upper alimentary tract in the rat:
Sensory ganglia and nuclei of the solitary and spinal trigeminal tracts. J.
Comp. Neurol. 283.248-268.
Arbab, M.A.R., T. Delgado, L. Wiklund, and N.A. Svendgaard (1988) Brain
stem terminations of the trigeminal and upper spinal ganglia innerva-
tion of the cerebrovacular system: WGA-HRP transganglionic study. J .
Cerebr. B1. Flow Metab. 835463.
Arvidsson, J. (1982) Somatotopic organization of vibrissae afferents in the
trigeminal sensory nuclei of the rat studied by transganglionic transport
of HRP. J. Comp. Neurol. 211384-92.
Arvidsson, J ., and S. Gobel (1981) An HRP study of the central projections of
primary trigeminal neurons which innervate tooth pulps in the cat.
Brain Res. 208:l-16.
Arvidsson, J., and E. Hellstrand (1988) A horseradish peroxidase study of
the central projections of the trigeminal neurons innervating the hard
palate in the cat. Brain Res. 451:197-204.
Astrom, K.E. (1953) On the central course of afferent fibers in the
trigeminal, facial, glossopharyngeal, and vagal nerves and their nuclei in
the mouse. Acta Physiol. Scand. 295209-320.
Beckstead, R., and R. Norgren (1979) An autoradiographic examination of
the central distribution of the trigeminal, facial, glossopharyngeal, and
vagal nerves in the monkey. J. Comp. Neurol. 184:455-472.
Boyle, R., and 0. Pompeiano (1981) Convergence and interaction of neck and
macular vestibular inputs on vestibulospinal neurons. J. Neurophysiol.
45:852-868.
Bratzlavsky, M. (1977) The connections between exteroceptive trigeminal
afferents and masticatory motoneurons in man. In Anderson and
Mathews (eds): Pain in the Trigeminal Region. Amsterdam: Elsevieri
North-Holland Biomedical Press, pp. 357-383.
Cechetto, D.F., D.G. Standaert, and C.B. Saper (1985) Spinal and trigeminal
dorsal horn projections to the parabrachial nucleus in the rat. J. Comp.
Neurol. 22:153-160.
Clarke, W., and D. Bowsher (1962) Terminal distribution of primary afferent
trigeminal fibers in the rat. Exp. Neurol. 6:372-383.
Contreras, R., R. Beckstead, and R. Norgren (1982) The central projections
of the trigeminal, facial, and glossopharyngeal and vagus nerves: An
autoradiographic study in the rat. J. Auton. Nerv. Syst. 63303-322.
Culberson, J.L., D.E. Haines, and P.B. Brown (1979) Contralateral projec-
tions of primary afferent fibers to mammalian spinal cord. Exp. Neurol.
64:83-97.
Darian-Smith, I. (1973) The trigeminal system. In A. Iggo (ed): Handbook of
Sensory Physiology, Vol. 11. Berlin: Springer-Verlag, pp. 271-314.
Erzurumlu, R., and H. Killackey (1979) Efferent connections of the brain-
stem trigeminal complex with the facial nucleus of the rat. J. Comp.
Neurol. 188:75-86.
Gobel, S., and J. Binck (1977) Degenerative changes in primary trigeminal
axons and in neurons in nucleus caudalis following tooth pulp extirpa-
tions in the cat. Brain Res. 132:347-354.
Gobel, S., W. Falls, and S. Hockfield (1977) The division of the dorsal and
ventral horns of the mammalian caudal medulla into eight layers using
anatomical criteria. In D.J. Anderson and B. Matthews (eds): Pain in the
Trigeminal Region. Amsterdam: ElsevierNorth Holland Biomedical
Press, pp. 443-453.
Halpern, B., and L. Nelson (1965) Bulbar gustatory responses to anterior
and to posterior tongue stimulation in the rat. Am. J. Physiol. 209:105-
110.
510
Hamilton, R., and R. Norgren (1984) Central projections of gustatory nerves
in the rat. J . Comp. Neurol. 222560-577.
Huerta, M., A. Frankfurter, and J . Harting (1983) Studies of the principal
sensory and spinal trigeminal nuclei of the rat: projections to the
superior colliculus, inferior olive, and cerebellum. J . Comp. Neurol.
220: 147-167.
Ikeda, M. (1979) Projections from the spinal and principal sensory nuclei of
the trigeminal nerve to the cerebellar cortex in the cat, as studied by
retrograde transport of horseradish peroxidase. J . Comp. Neurol. 184:
567-586.
J acquin, M., N. Chiaia, and R. Rhoades (1990) Trigeminal projections to
contralateral dorsal horn: central extent, peripheral origins, and plastic-
ity. Somatosens. Motor. Res. 7:153-183.
J acquin, M.F.R., W. Rhoades, H.L. Enfiejian, and M.D. Egger (1983a)
Organization and morphology of masticatory neurons in the rat: a
retrograde HRP study. J . Comp. Neurol. 218:239-257.
J acquin, M., K. Semba, M.D. Egger, and R. Rhoades (1983b) Organization of
HRP-labeled trigeminal mandibular primary afferent neurons in the rat.
J . Conip. Neurol. 215t397-420.
J acquin, M., K. Semba, R. Rhoades, and M.D. Egger (1982) Trigeminal
primary afferents project bilaterally to dorsal horn and ipsilaterally to
cerebellum, reticular formation, and cuneate, solitary, supratrigeminal,
and vagal nuclei. Brain Res. 246285-291.
J acquin, M., R. Stennett, W. Renehan, and R. Rhoades (1988) Structure-
function relationships in the rat brainstem subnucleus interpolaris: Low
and high threshold trigeminal primary afferents. J . Comp. Neurol.
267t107-130.
J erge, C.R. (1963) The function of the nucleus supratrigeminalis. J . Neuro-
physiol. 26:393402.
Kerr, F. (1963) The divisional organization of afferent fibres of the trigemi-
nal nerve. Brain. 86:721-732.
Kerr, F. (1972) Central relationships of trigeminal and cervical primary
afferents in the spinal cord and medulla. Brain Res. 43:561-572.
Kilduff, T., F. Sharp, and H. Heller (1983) Relative 2-deoxyglucose uptake of
the paratrigeminal nucleus increases during hibernation. Brain Res.
262117-123.
Kruger, L. (1971) Acritical review of theories concerning the organization of
the sensory trigeminal complex of the brainstem. I n K. Dubner and Y.
Kawamura (eds): Oral-Facial Sensory and Motor Systems. New York:
Appleton-Century-Crofts, pp. 135-158.
Kruger, L., S. Saporta, and S. Feldman 11977) Axonal transport studies of
the sensory trigeminal complex. In Anderson and Matthews (eds): Pain
in the Trigeminal Region. Amsterdam: ElsevierNorth-Holland Biomedi-
cal Press, pp. 191-201.
Light, A.R., and E.R. Perl, (1979) Reexamination of the dorsal root
projections to the spinal dorsal horn including observations on the
differential termination of coarse and fine fibers. J . Comp. Neurol.
186: 117-132.
Limwongse, V., and M. DeSantis (1977) Cell body locations and axonal
pathways of neurons innervating the muscles of mastication in the rat.
Am. J. Anat. 149:349444.
Lynch, R. (1985) A qualitative investigation of the topographical representa-
tion of masticatory muscles within the motor trigeminal nucleus of the
rat: A horseradish peroxidase study. Brain Res. 327:354-358.
Makous, W., S. Nord, B. Oakley, and C. Pfaffmann (1963) The gustatory
relay in the medulla. I n Y. Zotterman (ed): Olfaction and Taste. Oxford:
Pergamon Press, pp. 381-393.
Mantle-St J ohn, L., and D. Tracey (19871 Somatosensory nuclei in the
brainstem of the rat: independent projections to the thalamus and
cerebellum. J. Comp. Neurol. 255.259-271.
Marfurt, C.F. (1981) The central projections of trigeminal primary afferent
neurons in the cat as revealed by the transganglionic transport of
horseradish peroxidase. J. Comp. Neurol. 203: 785-798.
Marfurt, C.F., and C.E. Adams (1984) Transneuronal and transcellular
transport of horseradish peroxidase-wheat germ agglutinin (HRP-WGA)
in rat trigeminal sensory neurons. SOC. Neurosci. Abstracts 10:483.
Marfurt, C.F., and D.R. DelToro (1987) The corneal sensory pathway in the
rat: A horseradish peroxidase tracing study. J. Comp. Neurol. 261:450-
459.
Marfurt, C.F., and S.F. Echtenkamp (1988) Central projections and trigemi-
nal ganglion somatotopy of corneal afferent neurons in the monkey. J .
Comp. Neurol. 272:370-382.
Marfurt. C.F., and D.F. Turner (1983) Sensory nerve endings in the rat
orofacial region labelled by the anterograde and transganglionic trans-
port of horseradish peroxidase: A new method for tracing peripheral
nerve fibers. Brain Res. 261:l-12.
C.F. MARFURT AND D.M. RAJCHERT
Matesz, C. (1981) Peripheral and central distribution of fibers of the
mesencephalic trigeminal root in the rat. Neurosci. Lett. 27:13-17.
Matesz, C. (1983) Termination areas of primary afferent fibers of the
trigeminal nerve in the rat. Acta Biol. Hung. 34t3144.
Matsushita, M., M. Ikeda, and N. Okado (1982) The cells of origin of the
trigeminothalamic, trigeminospinal and trigeminocerebellar projections
in the cat. Neurosci. 7:1439-1454.
McKelvey-Briggs, D.K., J .A. Saint-Cyr, S.J. Spence, and G.D. Partlow 1989)
A reinvestigation of the spinovestibular projection in the cat using
axonal transport techniques. Anat. Embryol. 180:281-291.
Menetrey, D., J . Leah and J . DePommery (1987) Efferent projections of the
paratrigeminal nucleus in the rat. Neurosci. Lett. 73:48-52.
Mesulam, M.M. (1978) Tetramethylbenzidine for horseradish peroxidase
neurohistochemistry: A non-carcinogenic blue reaction-product with
superior sensitivity for visualizing neural afferents and efferents. Cy-
tochem. 26t106-117.
Miles, T.S., (1979) Features peculiar to the trigeminal innervation. Can. J .
Neurol. Sci. 6C2fi95-103.
Mizuno, N., A. Konishi, and M. Sat0 (1975) Localization of masticatory
motoneurons in the cat and rat by means of retrograde axonal transport
ofHRP. J. Comp. Neurol. 164:105-116.
Nazruddin, S., Y. Shirana, K. Yamauchi and Y. Shigenaga. (1989) The cells
of origin of the hypoglossal afferent nerves and central projections in the
cat. Brain Res. 490:219-235.
Neuhuber, W., and W. Zenker (1989) Central distribution ofcervical primary
afferents in the rat, with emphasis on proprioceptive projections to
vestibular, perihypoglossal, and upper thoracic spinal nuclei. J. Comp.
Neurol. 2801231-253.
Nomura, S., Y. Yasui, M. Takada, and N. Mizuno (1984) Trigeminal primary
afferent neurons projecting directly to the solitary nucleus in the cat: A
transganglionic and retrograde horseradish peroxidase study. Neurosci.
Lett. 50257-262.
Nord, S. (1967) Somatotopic organization in the spinal trigeminal nucleus,
the dorsal column nuclei and related structures in the rat. J. Cornp.
Neurol. 130:343-356.
Nord, S., and H. Kyler (1968) A single unit analysis of trigeminal projections
to bulbar reticular nuclei of the rat. J . Comp. Neurol. 134r485-494.
Nord, S., and G. Ross (1973) Responses of trigeminal units in the monkey
bulbar lateral reticular formation to noxious and non-noxious stimula-
tion of the face: experimental and theoretical considerations. Brain Res.
58:385-399.
Panneton, W., and H. Burton (1981) Corneal and periocular representation
within the trigeminal sensory complex in the cat studied with transgan-
glionic transport of horseradish peroxidase. J . Comp. Neurol. 199:327-
344.
Panneton, W.M., and H. Burton (1985) Projections from the paratrigeminal
nucleus and the medullary and spinal dorsal horns to the peribrachial
area in the cat. Neuroscience 15t779-797.
Pfaller, K., and J . Arvidsson (1988) Central distribution of trigeminal and
upper cervical primary afferents in the rat studied by anterograde
transport of horseradish peroxidase conjugated to wheat germ aggluti-
nin. J . Comp. Neurol. 268.9-108.
Phelan, K., and W. Falls (1989) The interstitial system of the spinal
trigeminal tract in the rat: anatomical evidence for morphology and
functional heterogeneity. Somatosens. Mot. Res. 6:367-399.
Porter, J. (1986) Brainstem terminations of extraocular muscle primary
afferent neurons in the monkey. J . Comp. Neurol. 247t133-143.
Pugh, W.W., and M. Kalia (1982) Differential uptake of peroxidase (HRP)
and peroxidase-lectin (HRP-WGA) conjugate injected in the nodose
ganglion of the cat. J . Histochem. Cytochem. 30:887-894.
Ram6n y Cajal, S. (1909) Histologie du Systeme Nerveux de 1Homme et des
Vertebres. Vol. 1. Paris: A. Maloine.
Rethelyi, M., D.L. Trevino and E.R. Perl. (1979) Distribution of primary
afferent fibers within the sacrococcygeal dorsal horn: An autoradio-
graphic study. J . Comp. Neurol. 185:603-622.
Rhoton, A., J . OLeary, and J. Paul Ferguson (1966) The trigeminal, facial,
vagal, and glossopharyngeal nerves in the monkey: afferent connections.
Arch. Neurol. 14:530-540.
Rob, J .T.M., J .D. van Willigen, and P.J .W. J iich (1986) Bilateral brainstem
connections of the rat supratrigeminal region. Acta Anat. 127:16-21.
Ruggiero, D., C. Ross, and D. Reis (1981) Projections from the spinal
trigeminal nucleus to the entire length of the spinal cord in the rat. Brain
Res. 225225-233.
Sessle, B., J . Hu, N. Amano, and G. Zhang (1986) Convergence of cutaneous,
tooth pulp, visceral, neck and muscle afferents onto nociceptive and
CENTRAL PROJECTIONS OF THE RAT TRIGEMINAL NERVE 511
non-nociceptive neurones in trigeminal nucleus caudalis (medullary
dorsal horn) and its implications for referred pain. Pain 27:219-235.
Shambes, G.M., J .M. Gibson, and W. Welker (1978) Fractured somatotopy in
granule cell tactile areas of rat cerebellar hemispheres revealed by
micromapping. Brain Bebav. Evol. 15:9&140.
Shigenaga, Y., I. Chen, S. Suemune, T. Nishimori, I. Nasution, A. Yoshida,
H. Sato, T. Okamoto, M. Sera, and M. Hosoi (1986a) Oral and facial
representation within the medullary and upper cervical dorsal horns in
the cat. J . Comp. Neurol. 243.388408.
Shigenaga, Y., T. Okamoto, T. Nishimori, S. Suemune, I. Nasution, I. Chen,
K. Yoshida, K. Tabuchi, M. Hosoi, and H. Tsura (1986b) Oral and facial
representation in the trigeminal principal and rostral spinal nuclei of the
cat. J . Comp. Neurol. 244:l-18.
Shigenaga, Y., S. Masayuki, T. Nishimori, S. Suemune, M. Nishimua, A.
Yoshida, and K. Tsuru (1988) The central projection of masticatoly
afferent fibers to the trigeminal sensory nuclear complex and upper
cervical spinal cord. J . Comp. Neurol. 268:489-507.
Smith, C. (1986) Sensory neurons supplying touch domes near the body
midlines project bilaterally in the thoracic spinal cord of rats. J . Comp.
Neurol. 245: 54 1-552.
Somana, R., N. Kotchabbakdi, and F. Walberg (1980) Cerebellar afferents
fromthe trigeminal sensory nuclei in the cat. Exp. Brain Res. 38:57-64.
Steindler, D.A. (1977) Trigemino-cerebellar projections in normal and reeler
mutant mice. Neurosci. Lett. 6:293-300.
Szentagothai, J . (1948) Anatomical considerations of monosynaptic reflex
arc. J . Neurophysiol. 1 1.445-454.
Takemura, M., T. Sugimoto, and A. Sakai (1987) Topographic organization
of central terminal region of different sensory branches of the rat
mandibular nerve. Exper. Neurol. 96:540-557.
Tiwari, R., and R. King (1974) Fiber projections fromtrigeminal nucleus
caudalis in primate (squirrel, monkey and baboon). J . Comp. Neurol.
158:191-206.
Torvik, A. (1956) Merent connections to the sensory trigeminal nuclei, the
nucleus of the solitary tract and adjacent structures. J . Comp. Neurol.
106.~51-141.
Vornov, J ., and J . Sutin (1983) Brainstem projections to the normal and
noradrenergically hyperinnervated trigeminal motor nucleus. J. Comp.
Neurol. 214: 198-208.
Watson, C., and R. Switzer, 111 (1978) Trigeminal projections to cerebellar
tactile areas in the rat-origin mainly from n. interpolaris and n.
principalis. Neurosci. Lett. 10:77-82.
Westrum, L., R. Canfield, and R. Black. (1976) Transganglionic degeneration
in the spinal trigeminal nucleus following removal of tooth pulps in adult
cats. Brain Res. I01t137-140.
Whitehead, M., and M. Frank (1983) Anatomy of the gustatory system in the
hamster: central projections of the chorda tympani and the lingual
nerve. J . Comp. Neurol. 220:378-395.
Yamamoto, T., R. Matsuo, and Y. Kawamura (1981) Localization of cortical
gustatory area in rats and its role in taste discrimination. J . Neurophys-
iol. 44:440445.