THE JOURNAL OF COMPARATIVE NEUROLOGY 371~325-335 (1996)

Transneuronal Changes of the Inhibitory

Circuitry in the Macaque Somatosensory
Thalamus Following Lesions
of the Dorsal Column Nuclei
HENRY J. RALSTON 111, PETER T. OHARA, XIAN WE1 MENG,
JOSEPH WELLS, AND DIANE DALY RALSTON
Department of Anatomy and the W.M. Keck Foundation Center for Integrative Neuroscience,
University of California, San Francisco, California 94143-0452 (H.J.R., P.T.O., X.W.M.,
D.D.R.); Department of Anatomy and Neurobiology, University of Vermont
Medical College, Burlington, Vermont 05405 (J.W.)

ABSTRACT
The inhibitory circuitry of the ventroposterolateral nucleus (VPL) of the macaque
somatosensory thalamus was analyzed in normal animals and in those surviving for a few days
or several weeks following a unilateral lesion of the cuneate nucleus, the source of medial
lemniscal (ML) axons carrying information from the contralateral upper extremity. Inhibitory
synaptic terminals in the VPL were defined as those that contain flattened or pleomorphic
synaptic vesicles and that can be shown to be immunoreactive for y-aminobutyric acid (GABA).
There are two types of these profiles: F axon terminals that arise from neurons of the thalamic
reticular nucleus, and perhaps from VPL local circuit neurons (LCNs); and the dendritic
appendages of LCNs that form presynaptic dendrites (PSDs). ML terminals normally have
extensive synaptic interactions with PSDs but not with F axon terminals. Electron microscopic
analyses revealed that cuneatus lesions resulted in a rapid loss of ML terminals and a
statistically significant reduction in both F and PSD synaptic profiles. Confocal scanning
microscopy also demonstrated a profound loss of GABA immunoreactivity in the deafferented
VPL. These changes persisted for more than 20 weeks, without any evidence of reactive
synaptogenesis of surviving sensory afferents or of inhibitory synapses. The changes in GABA
circuitry are transneuronal, and the possible mechanisms that may underlie them are
discussed. It is suggested that the altered GABAergic circuitry of the VPL in the monkey may
serve as a model for understanding changes in somatic sensation in the human following
peripheral or central deafferentation. D 1996 wiley-Liss, Inc.

Indexing terms: primate, GABA, electron microscopy, deafferentation, plasticity

S.L. Palay and G.E. Palade (1955) were the first to
demonstrate conclusively that there is a clear separation
between presynaptic and postsynaptic elements at chemical
synapses, thus confirming Cajal’s “neurone theory“ of the
organization of the nervous system. Despite enormous
strides taken in the study of the nervous system since that
time, analysis of neural organization by examining indi-
vidual synaptic profiles with the electron microscope still
remains a powerful technique. By using essentially the
same methods of the pioneering study of Palay and Palade
(19551, together with immunocytochemistry and scanning
confocal microscopy, we report here that central deafferen-
tation induces specific transneuronal changes in those
chemical synapses of the primate somatosensory thalamus
that utilize y-aminobutyric acid (GABA)as a neurotransmitter.
The ventroposterolateral nucleus ( W L ) of the primate
somatosensory thalamus contains two neuronal cell types
(as do most thalamic nuclei): 1)the thalamocortical relay
cell, usually about 20-40 pm in cell body diameter (Ohara
and Havton, 19941, which projects to functionally related
regions of the cerebral cortex and which constitutes approxi-
mately 75% of the total complement of neurons, and 2) the
interneuron (local circuit neuron-LCN), which represents
approximately 25% of the total neuronal population and
Accepted April 24, 1996.
Address reprint requests to Henry J. Ralston 111, Department of Anatomy,
S-1334, University of California, San Francisco, California, 94143-0452.
E-mail: hjr@phy.ucsf.edu

o 1996 WILEY-LISS, INC.

326
which exhibits y-aminobutyric acid immunoreactivity
(GABA-ir;Spreafico et al., 1983).When stained by the Golgi
method (Cajal, 1911; Tombol, 1969) or labeled by the
intracellular iontophoresis of horseradish peroxidase (HRP),
the interneurons have been shown to be morphologically
distinct from relay cells-being approximately 10-15 pm in
cell body diameter-and to possess complex dendritic ap-
pendages (Hamos et al., 1985). The thalamocortical relay
cells lack these dendritic appendages.
The primate W L receives two major ascending path-
ways, the medial lemniscus (ML) and the spinothalamic
tract (STT), which have partially overlapping terminal
domains within the VPL (Ralston and Ralston, 1992). ML
axons arise from the contralateral dorsal column nuclei and
convey information arising from non-noxious peripheral
stimuli, primarily mechanical. STT axons arise from neu-
rons of the contralateral spinal dorsal horn, and carry
information arising from a wide variety of noxious and
non-noxious mechanical, thermal, and chemical stimuli.
ML and STT axons terminate in the VPL as large presynap-
tic profiles (2-4 pm diameter), contain rounded synaptic
vesicles, and have therefore been termed RL (round vesicle,
large) axon terminals that form synapses with the VPL
neurons. Other classes of synaptic profiles in the VPL and
in other thalamic nuclei include RS profiles (round vesicles,
small, ca. 1-pm diameter), arising from cortical and brain-
stem neurons; and two GABA immunoreactive (GABA-ir)
profiles-F (flattened or pleomorphic vesicles) and PSD
(presynaptic dendrites; Ralston et al., 1988). F profiles are
the axon terminals of neurons of the thalamic reticular
nucleus (TRN), or the axons of interneurons. PSDs are the
vesicle-containing dendritic appendages of interneurons
(Ralston, 1971). All types of the axon terminals (RL, RS,
and F) form synapses with the dendrites of thalamic
neurons, but not with each other. PSDs, being of dendritic
origin, are postsynaptic to the three classes of axon termi-
nals and are presynaptic to the cell bodies and dendrites of
thalamic neurons (Ralston, 1991).
The RL-type medial lemniscal terminals contact the
dendrites of relay cells, as well as synapsing upon vesicle-
containing GABA-ir LCN dendritic appendages (Ralston
and Ralston, 1994) that, in turn, contact the relay cell
dendrites to form a triadic synaptic relationship. These
relationships between ML afferent, relay cell, and GABA-ir
appendages presumably serve a fundamental aspect of
information transfer in the thalamus, in which the afferent
axon transmits an excitatory message to the relay cell and
to the GABAergic appendages, resulting in feed-forward
GABAergic inhibition of the relay cell by the LCN dendritic
appendages (Pare et al., 1991). Although the substantial
majority ( > 80%) of lemniscal terminals form synaptic
contacts with GABA-ir presynaptic dendrites, most STT
terminals (about 85%)form simple axodendritic synapses
with relay cell dendrites, few having synaptic relationships
with GABA-ir presynaptic dendrites (Ralston and Ralston,
1994). Thus, there is little potential for immediate disynap-
tic GABAergic modulation of the STT activation of relay
cells.
Given the major synaptic interactions of ML terminals
with the GABA-ir dendritic appendages of LCNs, we wished
to determine whether there are demonstrable changes in
the GABA circuitry of the primate VPL following acute and
chronic lesions of the ML cells of origin in the dorsal column
nuclei. In this paper, we will show that there are profound
H.J. RALSTON I11 ET AL.
TABLE 1. Number of Terminal Profiles in Normal and Deafferented W L '
Profile
type Normal VPL 2-5 days 150 days
RL 0.937 f 0.211 0.214 i 0.064 0.15
(p < 0.051
F 1.44 f 0.092 1.044 f 0.093 0.74
(p < 0.051
PSD 2.27 i 0.394 0.996 f 0.147 1.27
(p < 0.051
RS 9.12 ? 0.459 10.844 f 0.615 11.52
(Dnot simificant)
'The number of profilesilO0 pm2 (mean and f s.e.m.) for each of the terminal types m
W L compared between 4 monkeys surviving 2-5 days after a lesion of the contralateral
nucleus cuneatus, and 3 normal animals. Average number of profiles counted per control
animal were RL-43; F-79, PSD-112; RS-435; Total of all types per control animal-668.
Average number of profiles counted per lesioned animal were: RL-4; F-22; PSD-22;
RS-207; Total of all types per lesion animal-255.
p = probability value cornpared to the normal group. Similar counts are given for a single
monkey 150 days followinga cuneatus lesion.
TABLE 2. Number of GABA-ir Profiles in Normal and Deafferented VPL'
Profile type Normal VPL 4 days 51 days
F 1.02 (1871 0.44 (96) 0.40 (1351
PSD 0.87 (1591 0.17 (36) 0.16 (551
'The numbers of GABA immunoreactive F axon terminals and presynaptic dendrites per
100 pm2 from a normal animal, and from animals surviving for 4 and 51 days after a
lesion of the contralateral nucleus cuneatus. The figures in parentheses give the number
of profiles counted. Although the numbers of the PSDs per unit area is less in the
immunoreacted tissue, the percent reduction in PSDs and F's in the deafferented
compared to normal W L is even greater than that seen in the unreacted tissue (Table 11.
See text for diseussion.
transneuronal reductions in both axonal and dendritic
GABA-ir synaptic profiles, F and PSD, respectively, as a
result of such deafferentation. The changes occur rapidly,
within a few days, and persist for at least 20 weeks. These
alterations in GABA innervation will be discussed in regard
to other investigations of thalamic plasticity, as well as the
possible changes of thalamic function that may occur as a
result of disordered thalamic circuitry.
MATERIALS AND METHODS
The 11 Mucucu fusciculuris monkeys in this study were
all adult males weighing 5-8 kg. They were housed and
cared for according to guidelines set forth by the Animal
Welfare Act and U.S.D.A. regulations, as well as by proto-
cols approved by the IACUC of the University of California,
San Francisco. Tissue other than that of the central
nervous system became part of the UCSF tissue sharing
program in support of other research. Thalamic tissue from
seven of the monkeys was not treated for GABA immunocy-
tochemistry and was used for a statistical analysis of
synaptic populations in the VPL (reported in Table 1).Of
these seven animals, three were normal and four survived
2-5 days after a unilateral lesion of the nucleus cuneatus
(see below), the results being compared with those of the
normals. An additional animal survived for 150 days after
cuneatus lesion. Thalamic tissue from the remaining three
monkeys was treated for GABA-ir and the results are listed
in Table 2. One of these animals received a unilateral
injection of tracer (WGA-HRP) into the cuneate nucleus 3
days prior to perfusion, the VPL from the ipsilateral
(unlabeled with HRP) and contralateral (HRP-labeled)
sides being used for synaptic counts; the other two ma-
caques survived for 4 or 51 days after a unilateral lesion of
the cuneate nucleus.
For surgical procedures, the monkeys were immobilized
with ketamine, 10 mg/kg, i.m., and anesthetized with
PLASTICITY OF THE PRIMATE THALAMUS 327

pentobarbital 1:l in saline i.v. titrated to an appropriate different for PSD characterization in the untreated tissue,
anesthetic level, followed by a maintenance i.v. dose of 5 in that a PSD needed to exhibit either a presynaptic or a
mgikgihour. Using sterile surgical procedures, unilateral postsynaptic contact. This difference may account for differ-
lesions of the nucleus cuneatus were made. Following the ent numbers of PSDs being counted in the GABA-ir com-
appropriate survival times, most animals were anesthetized pared to the untreated material (see results section).
and killed by the intracardiac perfusion of mixed aldehydes. For the analysis of the untreated VPL tissue, the Neuro-
The macaque that survived for 51 days was anesthetized, lucida (MicroBrightfield, Inc.) morphometric program was
and recordings were made from the VPL of both sides. used to determine the number of each terminal type per 100
Following characterization of the neuronal responses to pm2. Statistical comparisons were made with the student
cutaneous mechanical stimuli applied to the contralateral t test using Instat software. A finding was judged to be
upper extremity, extracellular microinjections (Pinault, significant if the P value was < 0.05.
1994) of biocytin were placed at the recording site, and the
anesthetized animal was then perfused with mixed alde- Confocal microscopy
hydes. Vibratome sections of the left and right VPL from the
In all animals, brainstem and thalamic blocks were cut animal surviving for 51 days after cuneate lesion were
serially on a Vibratome. The brainstem sections were processed by using Texas red as a fluorophore to identify
processed for confirmation of the extent of the experimental biocytin-labeled thalamic neurons, and a GABA antibody
lesion of the cuneate nucleus by using Nissl methods. The conjugated to fluorescein to demonstrate GABA-ir neuronal
thalamic sections were prepared for conventional electron cell bodies, axons, and dendrites. Series of single optical
microscopy (the first group of eight animals) or for electron sections, each about 1pm in thickness, were examined with
microscopic GABA immunocytochemistry (three animals). a BioRad dual beam scanning confocal microscope. The
Thalamic tissue in the monkey receiving thalamic microin- distribution of fluorescein-labeled GABA-ir appositions in
jections of biocytin was also processed for confocal scanning association with cell bodies and dendrites of the VPL
microscopy. The specificity of staining of structures for neurons were evaluated in the normal thalamus and in the
GABA immunoreactivity was confirmed by omission of the VPL, 5 1days following medial lemniscal deafferentation.
primary antibody, which resulted in an absence of staining. The confocal studies were compared with the electron
microscopic analyses of GABA-ir profiles in normal and
Electron microscopy deafferented animals. The findings for the structures in the
Selected regions of Vibratome sections of the VPL, normal VPL were used as a basis to compare changes in the
including ones adjacent to those used for confocal micro- distribution of synaptic profiles, whether untreated or
scopy, were prepared for electron microscopy. The regions processed for GABA-ir, following ML defierentation.
of the VPL studied were in the “core” of the nucleus to
which the medial lemniscus is known to project. In eight RESULTS
monkeys, standard methods were used to prepare the tissue
for thin-sectioning and subsequent analysis. In other ani- Examination of Nissl-stained transverse serial sections
mals, serial thin sections were processed immunocytochemi- through the medulla revealed that the lesions of nucleus
cally on the grid for the demonstration of GABA-ir by using cuneatus were nearly, or totally, complete, with few recog-
a modified postembedding technique with 10-nm gold par- nizable neurons appearing in the hemorrhagic (short sur-
ticles conjugated to the secondary antibody as previously vival) or gliotic (long survival) lesioned nucleus. Nissl-
described (Ralston and Ralston, 1994). stained sections of the microinjected thalamic tissue showed
Samples of normal and deafferented VPL, both the small numbers (usually 1-3) of biocytin-labeled neurons in
standard sections and those treated for GABA, were counted the more medial regions of the W L , neurons of this zone
to assess the number of synaptic profiles per 100 pm2. For having responded to cutaneous stimuli applied to the
the analysis of synaptic profile types, two methods were contralateral arm and hand.
used. 1)Micrographs from nonoverlapping fields of the core
region of the VPL were taken at random intervals. From Electron microscopy
these micrographs, all profiles were identified (see below) Nonimmunoreacted tissue. The samples of VPL from
and counted, the counts being expressed as profile numbers these animals were not reacted for GABA-ir. In counts of
per 100 pm2. 2) Profiles were identified and counted several hundred synaptic profiles of each type, the numbers
directly from the electron microscope viewing screen as of each type per unit area in control animals fell within a
nonoverlapping regions of the specimen were traversed narrow range (Table 2). After lesions of the contralateral
across the viewing field. By measuring the width and length cuneate nucleus, the number of RL profiles was reduced by
of the region viewed, profile counts were expressed as more than 75% in the 2-5-day period following the lesion.
number per 100 pm2. The synaptic profiles were designated There was also a statistically significant reduction in the F
as follows: profiles with rounded synaptic vesicles were and PSD profiles in the first few days following the lesion.
classified as RL (ascending sensory afferent type) or RS The numbers of RS profiles did not change significantly at
(from cortex or brainstem). Those containing flattened or any time interval after cuneate lesion. The counts of all
pleomorphic synaptic vesicles were designated as being of synaptic profile types in the VPL from the 150-day survival
axonal (F) or dendritic (PSD) origin, respectively. Synaptic animal were within the ranges found in the 2-5 day
vesicles fill the F profiles, but only occupy a small portion of operated animals, indicating that the changes found at the
the PSDs, which are both presynaptic and postsynaptic short survival period were maintained for several months.
(Ohara et al., 1989), in contrast to F profiles, which are The results of these analyses are shown in Table 1.
exclusively presynaptic elements. The identification of PSDs Tissue reacted for GABA immunocytochemistry. The
made in the GABA-ir material required that the profile be postembedding GABA immunocytochemical technique
postsynaptic in a given section. The criteria were somewhat yields clearly stained structures, the gold particles being
328
present in certain profiles at densities > 5 times that of
background, the latter measured over vascular elements.
The GABA-ir synaptic profiles were characterized, classi-
fied, and counted, the results being expressed as numbers
per 100 Fm2. F profiles were judged to be axon terminals;
they are filled with flattened or pleomorphic vesicles,
contain one or more mitochondria, and are only presynaptic
to other thalamic elements (Figs. 1, 2). PSDs (presynaptic
dendrites) have rounder and more widely scattered synaptic
vesicles than seen in F profiles. PSDs are both postsynaptic
to all presynaptic types in the VPL (Figs. 3-5) as well as
being presynaptic to neuronal cell bodies, dendritic shafts,
and to other PSDs.
Counts of several hundred GABA-ir profiles in the nor-
mal VPL revealed about 1.0 GABA-ir axon terminals (F)
and 0.9 GABA-ir PSDs per 100 pm2. The counts of F
profiles per unit area were about the same as those of the
normal unreacted tissue (Compare Tables 1 and 2). The
number of GABA-ir PSDs per unit area were less than half
those counted in the unreacted tissue. Perhaps this differ-
ence may be accounted for by the different criteria used to
identify a profile as a PSD in the GABA-ir compared to the
nonreacted tissue (see discussion). Following cuneate
nucleus lesions at both survival times studied, short (4
days) and long (51days), counts of several hundred GABA-ir
profiles in the deafferented VPL revealed that both GABA-ir
profile types, axon terminals (F), and presynaptic dendrites
(PSDs), were reduced by more than 50%. The counts of
GABA-ir profiles are given in Table 2.
Confocal microscopy
The extracellular microinjection method (Pinault, 1994)
resulted in small numbers of well-labeled individual VPL
neurons with well-defined dendritic arbors. In our prepara-
tions, the red fluorescence is that of the biocytin-stained
thalamic neurons labeled with Texas red. The green fluores-
cence is that of the GABA antibody conjugated to fluores-
cein. GABA-ir structures were abundant within the first
12-15 pm of the surface of a given 50-pm section, but the
staining of GABA-ir thalamic elements diminishes rapidly
at about 15-20 pm from the surface, most likely due to the
reduced penetration of the GABA antibody at deeper levels
of the tissue. Therefore, our analyses concentrated on
optical sections within the first 12 pm of the surface. In
single optical sections of the normal VPL, as well as in the
merged images of many sections, there is a dense network
of GABA-ir fluorescent structures, many in close apposition
(yellow) to the somata and dendrites of VPL neurons (Fig.
6). In contrast, the density of GABA-ir fluorescent struc-
tures in single and merged sections were markedly reduced
in the VPL following medial lemniscal deafferentation
(Fig. 7).
DISCUSSION
The principal electron microscopic findings of the present
study are that two major inhibitory synaptic elements of
the VPL are substantially reduced in number by lesions of
the principal sensory afferent to the VPL, the medial
lemniscus. Both of these synaptic profile types, axon termi-
nals (F) and presynaptic dendrites (PSDs), are well labeled
by GABA immunocytochemistry. The reductions in their
numbers per unit area occur within a few days of deafferen-
tation, and their numbers remain reduced for at least
several months. GABA-ir axons arise from neurons of the
H.J. RALSTON 111 ET AL.
thalamic reticular nucleus (TRN) and from local circuit
neurons (Penny et al., 1983; Spreafico et al., 1983; Ohara et
al., 1989). GABA-ir dendrites and dendritic appendages are
processes of intrinsic local circuit neurons (LCNs) of the
VPL. Thus, in these macaques, partial somatosensory
deafferentation (lesion of the cuneate nutleus) induces
substantial transneuronal changes in GABA-ir circuitry,
leaving the population of RS type synapses arising from the
cortex or brainstem unaffected.
We used two types of quantitative analyses to evaluate
the changes in the VPL following lesions of the cuneate
nucleus. First, in the tissue that was not processed for
GABA-ir, we counted the numbers of synaptic profiles in
three control and in four short-term survival animals. The
results of these counts were then analyzed statistically
(Table 1). The other analysis was based on counts of
GABA-ir profiles (F and PSD) in one normal, one short-
term, and one long-term survival animal. The principal
difference between the results of the two methods was the
lower numbers of PSDs in the GABA-ir tissues, both
control and postoperative. However, the criteria for classify-
ing a profile as being a PSD was somewhat different in the
two methods. In the nonimmunoreacted tissue, a profile
was designated as a PSD if it had scattered pleomorphic
synaptic vesicles and was either pre- or postsynaptic. In the
GABA-ir tissue, a profile had to be postsynaptic in a single
section to be designated as a PSD. This latter method
undoubtedly resulted in an underestimate of the number of
PSDs in both normal and experimental tissues, as the
length of presynaptic appositions (see Figs. 3-5) is clearly
less than the diameter of the PSD, so that many sections
would not pass through the plane of the presynaptic
contact. The reason for the different criteria used was that
in the nonimmunoreacted tissue, the vesicle shape, size,
and packing density that characterizes PSDs are more
readily apparent. In the GABA-ir tissue, the fine-structural
features are more difficult to define, so that we required a
GABA-ir profile to be postsynaptic in order to classify it as a
PSD. Nonetheless, both methods of analysis had the same
result: lesions of the ML afferent system lead to substantial
transneuronal reductions in the inhibitory circuitry of the
VPL.
As pointed out by Coggeshall and Lekan (19961, the type
of analysis carried out here cannot determine absolute
numbers of synaptic terminals. On the other hand, this
analysis can provide information t o compare the counts of
synaptic profile density between the normal and the experi-
mental animals if there are no substantial changes in the
volume of the VPL in the two groups of animals. Qualita-
tive analysis of the tissue did not reveal any evidence of
degeneration of thalamic neurons or of reactive gliosis.
Quantitative analysis of the density of RS synaptic profiles
following cuneate lesions revealed no significant changes
from the normal, further indicating that there was no
substantial change in the VPL volume as a result of the
deafferentation. We conclude that the changes in the
Figs. 1, 2. y-Aminobutyric acid immunoreactive (GABA-ir) axon
terminals (F) in the ventroposterolateral nucleus ( W L ) , contacting
(arrows) dendritic shafts (D). The profiles were judged to be of the F
type because of the high packing density of the synaptic vesicles and the
fact that such profiles are never found to be postsynaptic (see text).
Many of these terminals arise from the thalamic reticular nucleus;
others might be axons of local circuit neurons. Scale bars = 1 Fm.
Figures 1 and 2
 Figs. 3, 4. Electron micrographs. GABA-ir presynaptic dendrites Fig. 4. A round vesicle, large (RL) type profile characteristic of sensory
(PSDs). Note the scattered synaptic vesicles and electron-lucent cyto- afferent projections to thalamic nuclei contacts a GABA-ir presynaptic
plasm that characterize PSDs. Fig. 3. A small GABA-ir PSD is dendrite at the arrow. Scale bars = 1pm.
postsynaptic (arrow) to an axon terminal (round vesicle, small; RS).
PLASTICITY OF THE PRIMATE THALAMUS
Fig. 5. A PSD is postsynaptic (dark arrow) to a small axon terminal (RS) and exhibits multiple
filamentous contacts with a neuronal cell body. Scale bar = 1pm.
density of GABA-ir axon terminals and presynaptic den-
drites were a result of the experimental lesion, rather than
a change in the “reference space” (Coggeshall and Lekan,
1996).
Confocal microscopy provides an excellent overview of
fluorescently labeled structures in the monkey VPL. The
dendritic arbors of Texas red-labeled neurons extend
throughout the thickness of the 50-pm vibratome sections.
However, GABA-ir fluorescence only extends for about 15
pm from the section surface, probably due to limited
penetration of the GABA antibody. Given this caveat, the
findings by confocal microscopy are in keeping with those
by electron microscopy: cuneate nucleus lesions lead to a
transneuronal reduction of GABA-ir structures in the
deafferented VPL.
Why do the transneuronal changes occur?
We can only speculate upon the mechanisms underlying
the rapid reductions in both F axon terminals and in
presynaptic dendrites following ML lesions. We have previ-
ously shown that ML axon terminals commonly synapse
upon PSDs, as well as upon the dendritic shafts of projec-
tion neurons (Ralston and Ralston, 1994). Therefore, it is
331
quite possible that the PSDs, which are the dendritic
appendages of local circuit neurons, retract rapidly follow-
ing loss of one of their principal synaptic inputs. The PSDs
apparently do not recover, and neither do the numbers of
the RL terminals of medial lemniscus axons, in contrast to
the reactive synaptogenesis that has been reported in the
rat VPL following ML deafferentation (Wells and Tripp,
1987).
The reasons for changes in the F axon terminal type are
more difficult to understand, because many, and perhaps
most, F axon terminals are those of thalamic reticular
neurons (TRN). The TRN does not receive ascending
sensory afferent input, and it is unlikely that TRN neurons
would be rapidly affected by ML deafferentation. However,
a fraction of F terminals might arise from local circuit
neurons (LCNs), and the axons of these cells may also
retract as a consequence of the loss of ML input to the
dendritic arbors of the neurons. This explanation would
require that the axons of LCNs are of the F type, and there
is no evidence at present that this is the case. Another
possibility is that F terminals that are near ML afferents
are somehow affected by the loss of ML synapses. Our
three-dimensional reconstructions of ML afferents in the
 Fig. 6. A confocal microscopic image of a labeled projection neuron
(red) from the normal macaque W L . In this 1-pm optical section taken
4 pm from the section surface a large number of GABA-ir (green)
labeled profiles can be seen. The projection neuron soma and primary
dendrite are studded with GABA-ir profiles and close appositions
(yellow) are frequent (e.g., arrows). An asterisk (*) identifies the cell
body of a GABA-ir local circuit neuron. Inset: A merged image of
sixteen 1-Fm optical sections which further illustrates the relationship
between the labeled neurons and GABA-ir profiles.
 Fig. 7. A confocal microscopic image of labeled neurons in the W L arrows. The amount of GAl3A-ir fluorescence is substantially reduced
following a lesion of the contralateral cuneate nucleus 51 days earlier. when compared to normal VPL (Fig. 6). Inset: A merged image of
The section is a 1-pm optical section taken 6 pm from the surface. Some thirteen 1-Fm optical sections.
of the presumed synaptic appositions (yellow) are indicated by the
334 H.J. RALSTON I11 ET AL.

VPL (Ralston and Ralston, 1994) revealed that a given representation of the extremity, so that only non-hairy skin
segment of the relay cell dendritic arbor that received is represented, but two months later a topographically
labeled ML terminals also exhibited nearby synaptic con- appropriate representation of the hairy-skin reappears in
tacts from F axon terminals. A loss of the ML input to a the appropriate region of the thalamus (Garraghty and
dendritic segment may cause retraction of F terminals Kaas, 1991). 4) In the raccoon, both reversible and irrevers-
contacting the same segment of dendrite. Other possible ible denervation of a single digit results in the immediate
causes of reductions in the GABA-ir circuitry following unmasking of inhibitory responses of the deafferented cells
deafferentation include altered activity of thalamic neu- by stimulation of adjacent, normally responding digits,
rons, or changes in the synthesis of GABA or GABA which is similar to that reported in the cortex (Rasmusson
receptors. The fact that such transneuronal changes occur et al., 1993). 5 ) Transient blocking of facial whisker affer-
rapidly is both interesting and intriguing, but we do not ents by injection of anesthetic in the rat results in an
understand the mechanisms involved. immediate unmasking of short latency responses of the
neurons in the ventroposteromedial nucleus (VPM) within
The function of GABAergic the anesthetized zone, and these neurons then become
circuitry in the VPL responsive to adjacent, non-anesthetized regions (Nicolelis
Whatever the reasons for the reductions in the numbers et al., 1993). 6 ) Injuries of the spinothalamic tract (STT) are
of F axon terminals and of PSDs, one would expect substan- believed to play a seminal role in the genesis of central
tial changes in the functions of the surviving elements of post-stroke pain in humans (Boivie et al., 1989). Patients
the VPL circuitry as a consequence of these losses. Pare et exhibiting this pain syndrome have been found to have
al. (1991) have examined the interactions of mammillary abnormal bursting responses of thalamic neurons (Lenz et
body afferent axons (analogous to the ML afferents of the al., 1989), which mirror neuronal hyperactivity demon-
VPL) with relay cells and presynaptic dendrites in the cat strated in the cat with STT lesions. Following anterolateral
anterior nuclear complex. They described a triphasic series cordotomy in the cat, VPL cells have been shown to exhibit
of inhibitory postsynaptic potentials (IPSPs) in relay cells hyperactive responses to both innocuousand noxious periph-
following stimulation of mammillothalamic afferents: a eral stimulation, and these responses can be blocked by
short-latency, brief-duration IPSP (designated “a”) that MK-801, an antagonist of the NMDA receptor (Koyama et
was determined to be due to feedforward inhibition medi- al., 1993).
ated by the triadic synaptic relationships between mammil- Common themes in the studies cited in the previous
lary afferents, interneuronal presynaptic dendrites, and paragraph are that peripheral or central dederentation
relay cells; and two-longer latency, more prolonged IPSPs induces thalamic neurons to become more excitable, to
(designated A and B) that were assumed to be evoked by exhibit abnormal bursting behaviors, and to enlarge the
GABAergic axon terminals. It was shown that a and A were sizes of their receptive fields. Given the role of GABA-
mediated by GABAA receptors, and that the B IPSP was due mediated inhibition in thalamic functioning, it is quite
to activation of GABAB receptors. They suggested that the possible that alterations in GABAergic circuitry similar to
short-duration IPSPs in relay cells resulting from afferent those described in the present report underlie the changes
activation of presynaptic dendrites would promote effective in thalamic functions followingdeafferentation.
information transfer by the relay cells by reducing postsyn- In summary, partial deafferentation of the somatosen-
aptic summation, leading to the rapid return of the relay sory thalamus by lesions of the dorsal column-medial
cell to its resting potential to permit a higher frequency of lemniscus system leads to rapid, substantial, and sustained
information transfer. Assuming that the findings of Par6 et transneuronal loss of the innervation of the VPL projection
al. (1991) are applicable to the primate somatosensory cells by GABAergic axons and dendrites. Although the
thalamus, it is likely that the information transfer between mechanisms underlying these transneuronal changes are
surviving afferents (mostly spinothalamic tract axons) and not understood, they likely would have a profound effect on
relay cells, with reduced modulation by GABAergic presyn- information transfer by the surviving elements of the VPL
aptic dendrites or F axon terminals, is substantially differ- circuitry. It is possible that plastic changes described here
ent than that evoked by stimulation of the ML in the in the macaque are analogous to deafferentation injuries to
normal thalamus. the central nervous system in humans, such as that occur-
Previous studies of thalamic plasticity ring in “central post-stroke pain.” We are now seeking to
discover whether macaque monkeys having dederentation-
Other studies have shown that the somatosensory thala- induced alterations of thalamic circuitry have demon-
mus exhibits reorganization following acute or chronic strable changes in behavior, as measured by abnormal
deafferentation. 1) In an examination of macaques that responses to innocuous cutaneous stimuli. A behavioral
survived more than 12 years after extensive dorsal rhi- correlate in the monkey may provide some understanding
zotomy, the investigators reported a profound transneuro- of the possible changes in neuronal circuitry associated
nal degeneration of the ipsilateral dorsal column nuclei, and with abnormal sensory experiences in humans.
a loss of axons and neurons staining for the calcium-binding
protein, parvalbumin, but not of calbindin 28-kDa, and a
marked decrease of GABAA receptors in the thalamus ACKNOWLEDGMENTS
contralateral to the deafferented limb (Rausell et al., 1992).
2) A lesion of the gracile nucleus, carrying information from We thank Ms. Antonia M. Milroy for her electron micro-
the hindlimb, results in an expansion of the representation scopic and GABA immunocytochemical preparations, Ms.
of the forelimb into the deafferented thalamic zone in the Sandra A. Canchola for her histological and photographic
macaque (Pollin and Albe-Fessard, 1979). 3) Denervation of materials, and Ms. Jenny Wong for her assistance in data
the hairy-skin of the distal extremity of the squirrel mon- base maintenance. This work was supported by grant NS
key results in a transient reorganization of the thalamic 23347 from the National Institutes of Health.
PLASTICITY OF THE PRIMATE THALAMUS
LITERATURE CITED
Boivie, J., G. Leijon, and I. Johansson (1989) Central post-stroke pain-a
study of the mechanisms through analyses of the sensory abnormalities.
Pain 37t173-185.
Cajal, S.R. (1911) Histologie du Systeme Nerveux de l'Homme et des
Vertehres. Val. 11. Paris: L. Azoulay, Figs. 254, 260. (Reprinted Edition,
1955).
Coggeshall, R.E. and H.A. Lekan (1996) Methods for determining numbers
of cells and synapses: A case for more uniform standards of review. J.
Comp. Neurol. 3643-15.
Garraghty, P.E. and J.H. Kaas (1991) Functional reorganization in adult
monkey thalamus after peripheral nerve injury. NeuroReport 2747-
750.
Hamos, J.E., S.C. Van Horn, D. Raczkowski, D.J. Uhlrich, and S.M.
Sherman (1985) Synaptic connectivity of a local circuit neurone in lateral
geniculate nucleus of the cat. Nature 317t618-621.
Koyama, S., Y. Katayama, S. Maejima, T. Hirayama, M. Fujii, and T.
Tsubokawa (1993) Thalamic neuronal hyperactivity following transec-
tion of the spinothalamic tract in the cat: Involvement of N-methyl-D-
aspartate receptor. Brain Res. 612345-350.
Lenz, F.A., H.C. Kwan, J.O. Dostrovsky, and R.R. Tasker (1989) Character-
istics of the bursting pattern of action potentials that occurs in the
thalamus of patients with central pain. Brain Res. 496357-360.
Nicolelis, M.A.L., R.C.S. Lin, D.J. Woodward, and J.K. Chapin (1993)
Induction of immediate spatiotemporal changes in thalamic networks by
peripheral block of ascending cutaneous information. Nature 361t533-
536.
Ohara, P.T., G. Chazal, and H.J.111 Ralston (1989) Ultrastructural analysis
of GABA-immunoreactive elements in the monkey thalamic ventrobasal
complex. J. Comp. Neurol. 283t542-558.
Ohara, P.T. and L.A. Havton (1994) Preserved features of thalamocortical
projection neuron dendritic architecture in the somatosensory thalamus
of the rat, cat and macaque. Brain Res. 648359-264.
Palay, S.L. and G.E. Palade (1955) The fine structure of neurons. J. Biophys.
Biochem. Cytol. 1:69-88.
Pare, D., R.C. Dossi, and M. Steriade (1991) Three types of inhibitory
postsynaptic potentials generated by interneurons in the anterior tha-
lamic complex of cat. J. Neurophysiol. 66t1190-1204.
335
Pinault, D. (1994) Golgi-like labeling of a single neuron recorded extracellu-
larly. Neurosci. Lett. 170255-260.
Penny, G.R., D. Fitzpatrick, D.E. Schmechel, and I. T. Diamond (1983)
Glutamic acid decarboxylase-immunoreactive neurons and horseradish
peroxidase-labeled projection neurons in the ventral posterior nucleus of
the cat and galago senegalensis. J. Neurosci. 3t1868-1887.
Pollin, B. and D. Albe-Fessard (1979) Organization of somatic thalamus in
monkeys with and without section of dorsal spinal tracts. Brain Res.
173t431449.
Ralston, H.J.III(1971) Evidence for presynaptic dendrites and a proposal for
their mechanism of action. Nature 23Ot585-587.
Ralston, H.J.III(1991) Local circuitry of the somatosensory thalamus in the
processing of sensory information. Prog. Brain Res. 87: 13-28.
Ralston, H.J.111, P.T. Ohara, D.D. Ralston, and G. Chazal (1988) The
neuronal and synaptic organization of the cat and primate somatosen-
sory thalamus. In M. Bentivoglio and R. Spreafico (eds): Cellular
Thalamic Mechanisms. New York: Elsevier, pp. 127-141.
Ralston, H.J.111 and D.D. Ralston (1992) The primate dorsal spinothalamic
tract: Evidence for a specific termination in the posterior nuclei (PolSG)
of the thalamus. Pain 48t107-118.
Ralston, H.J.111 and D.D. Ralston (1994) Medial lemniscal and spinal
projections to the Macaque thalamus: An electron microscopic study of
differing GABAergic circuitry serving thalamic somatosensory mecha-
nisms. J. Neurosci. 14:2485-2502.
Rasmusson, D.D., D.F. Louw, and S.A. Northgrave (1993) The immediate
effects of peripheral denervation on inhibitory mechanisms in the
somatosensory thalamus. Somatosens. Mot. Res. lOt69-80.
Rausell, E., C.G. Cusick, E. Taub, and E.G. Jones (1992) Chronic deafferen-
tation in monkeys differentially affects nociceptive and nonnociceptive
pathways distinguished by specific calcium-binding proteins and down-
regulates gamma-aminobutyric acid type A receptors at thalamic levels.
Proc. Natl. Acad. Sci. USA 89:2571-2575.
Spreafico, R., D.E. Schmechel, L.C. Ellis, and A. Rustioni (1983) Cortical
relay neurons and interneurons in the N.ventralis posterolateralis of
cats: A horseradish peroxidase, electron microscopic, golgi and immuno-
cytochemical study. Neuroscience 9r491-509.
Tombol, T. (1969) Two types of short axon (golgi 2nd) interneurons in the
specific thalamic nuclei. Acta Morpholog. Hung. 17.285-297.
Wells, J. and L.N. Tripp (1987) Time course of reactive synaptogenesis in the
subcortical somatosensory system. J. Comp. Neurol. 255:46&475.