Neuroscience Vol. 27, No. 3, pp. W-863, 1988 0306~4522/88 $3.00 + 0.

00
Printed in Great Britain Pergamon Press plc
0 1988IBRO

ASSOCIATION OF DOPAMINE D, AND D, RECEPTORS

WITH SPECIFIC CELLULAR ELEMENTS IN THE BASAL
GANGLIA OF THE CAT: THE UNEVEN TOPOGRAPHY
OF DOPAMINE RECEPTORS IN THE STRIATUM IS
DETERMINED BY INTRINSIC STRIATAL CELLS,
NOT NIGROSTRIATAL AXONS

R. M. BECKSTEAD
Department of Anatomy and Cell Biology, Medical University of South Carolina, Charleston, SC 29425,
U.S.A.

Abstract-To ascertain the cellular associations of the D, and D, dopamine receptor subtypes in
components of the basal ganglia, cats were prepared with unilateral, axon-sparing, ibotenic acid lesions
of the striatum (n = 6) or lesions of the nigrostriatal dopamine system by intranigral infusion of
6-hydroxydopamine (n = 8). After 42 days survival, tissue sections from the brains were processed for
quantitative, in oitro receptor autoradiography with [H]SCH23390 (D, radioligand) or [Hlspiroperidol
(D2 radioligand). Lesion-induced changes in basal ganglia nuclei were assessed by comparing them to the
corresponding nuclei on the intact side and in naive brains.
Ibotenate lesions cause a decline in specific D, and D, receptor-binding in the area of the striatal lesion
of 94% and 85%, respectively, and completely eliminate the uneven patterns of high- and low-density
binding that are characteristic of the cats caudate nucleus. The globus pallidus, entopeduncular nucleus
and pars reticulata of the substantia nigra also show marked reductions in binding after striatal ibotenate
lesions. Thus, after caudate nucleus lesions, D, binding in the two pallidal segments declines by
approximately 50%, but remains unchanged in the substantia nigra. Binding of the D, radioligand (which
is not measurable in the globus pallidus) declines by about 75% in the affected regions of the
entopeduncular nucleus and pars reticulata, and by about 30% in the pars compacta.
Lesions of the nigral dopamine neurons reduce D, receptor-binding by 95% in the pars compacta and
40% in the pars reticulata, but have no effect on the concentration of D, or Dr radioligand-binding in
the striatum or pallidum. Moreover, such lesions failed to alter the uneven patterns of binding in the
striatum.
These data suggest that most, if not all, D, receptors in the basal ganglia are associated with cells of
the striatum and their axons in the entopeduncular nucleus and substantia nigra, and likewise, a large
majority of D, receptors are associated with striatal cells and their axons in pallidal structures. Nearly
all D, receptors in the substantia nigra are associated with dopamine neurons (autoreceptors). Finally,
the heterogeneous patterns of D, and D, receptors in the striatum are a consequence of intrinsic neuronal
distributions.

Dopamine transmission in the basal ganglia can be
mediated by either of two dopamine receptor sub-
types termed D, and D,.34 Since the observation that
activation of the D, subtype produces an increase in
adenylate cyclase activity while activation of the D,
subtype depresses this enzyme activity,4,43*57*61 much
evidence has accumulated which indicates that the D,
and D, receptor subtypes mediate different, and
sometimes interactive, biological responses to do-
pamine transmission (for discussion see Ref. 13).
Because of these functionally significant relationships
of the two types of dopamine receptor, it has been
considered important to identify their regional distri-
Abbreviafions: SCH23390, (R) - (+) - 8 - chloro - 2,3,4,5 -
tetrahydro-3-methyl-5-phenyl-IH-3-benzazepin-7-01;
SKF38393, 2,3,4,5-tetrahydro-7,8-dihydroxy-l-phenyl-
IH-3-benzazepine.
butions in the basal ganglia, especially with respect to
known features of anatomical and neurochemical
organization.
Two features of the anatomical-chemical or-
ganization of the basal ganglia, which have emerged
from the convergence of independent experimental
evidence, are the compartmental organization of the
striatum and the dual nature of the striatopallidal-
nigral projection system, both of which are described
by a number of cytochemical and connectional char-
acteristics (for refs see Ref. 8). Several receptor-
binding studies in mammals have suggested that the
topographic distributions of D, and Dz dopamine
receptors may be highly relevant to these frame-
works. Thus, receptor-binding in rats has shown that
both types of dopamine receptor are distributed in a
high-to-low, lateromedial gradient within the
caudatoputamen2~8~9~52-a pattern which has been

851
852 R. M. BECKSTEAD

likened to the distribution of acetylchohnesterase-
and the receptors are not equally present in pallidal
and nigral structures.0~6~48~52
A more intricate, patchy pattern of high and low
density of D, and D, receptors has been found in the
the dopamine cell populationsh After 6 weeks survival,
ail cats were deeply re-anesthetized with pentobarbital
(Nembutal), and their brains were removed, immediately
frozen in dry ice, and stored overnight at -70C.
Binding procedures

striatum of the cat,8,47.4*and it has been noted further The binding procedure was carried out as described
previously. Briefly, five adjacent frontal sections of 20 pm
that the two receptor-binding patterns are at least thickness were cut at intervals of 200 pm on the cryotome,
partially complementary in the caudate nucleus. and thaw-mounted onto poly-o-lysine-coated slides. After
Moreover, the high density zones of D, receptor- air-drying for 30 min at 22C the tissues were immersed for
binding are comparable in size, shape, and location IO min in ice-cold, 0.01 M sodium phosphate-buffered saline
(pH 7.4). The binding incubations were carried out in the
to the dense terminal zones of the uneven nigro-
same buffer solution using the first series of sections for
striatal projection system and the coincident sub- non-specific [Hlspiroperidol-binding, the second series for
stance P cell clusters described for the cat. The total [3H]spiroperidol-binding, the third series for total
dissociation of the two receptor subtypes in palhdal [ZH]SCH23390-binding, and the fourth for non-specific
and nigral structures also appears to be more pro- [rH]SCH23390-binding. The fifth series was used for stain-
ing with Cresyl Violet. The concentration of radioligand
nounced in the cat such that no D, receptors appear chosen was based on published KD (dissociation constant)
to be present in the globus pallidus at all while they values for the cat. These values for [Hlspiroperidol and
are abundant in the entopeduncular nucleus and for [rH]SCH23390 are. respectively, 0.25 nM and 0.55 nM.
substantia nigra. 8.48 Conversely, D, receptors are Both [Hlspiroperidol and [H]SCH23390 were obtained
from New England Nuclear (specific activities = 22.5
more dense in the globus pallidus than in the ento-
and 80.4 Ci/mmol, respectively). Determination of total
peduncular nucleus and substantia nigra.8,48 binding was done under the following conditions: for
No attempt has been made to determine the cellu- [ZH]spiroperidol, the incubation buffer solution contained
lar associations of the two receptor subtypes sepa- 2 nM radioligand, 2 PM cinanserin (to block type 5HT,
rately in the globus pallidus, entopeduncular nucleus, serotonin receotors). and 2uM SCH23390 (to block D,
receptors), and iasted for 30min at . 22C; for
and substantia nigra in a non-rodent species. More- [H]SCH23390, the incubation buffer solution contained
over, because the pattern of D, and D, receptor- 2nM radioligand, 2pM cinanserin, and 2pM
binding in the cat caudate nucleus is reminiscent of (-)-sulpiride, and lasted for 90 min at 37C. At these
the uneven terminal pattern of the nigrostriatal radioligand concentrations (at least four times the KD
values), a fractional occupancy of 8&90% is obtained.
projection, it seems possible that the dense zones of
Non-specific binding was determined for both radioligands
D2 dopamine receptors are a consequence of pre- by competition with 2 p M (+)-butaclamol. The incubations
synaptic receptors in nigral axons. For these reasons, were terminated by two 10 s rinses in ice cold buffer and a
we sought first to determine whether the hetero- 5 s rinse in distilled water. The tissue sections were quickly
geneous patterns of D, and D2 receptor distributions dried under a stream of cool air, apposed to LKB Ultrofilm
along with plastic tritium microscales (Amersham) in X-ray
in the cat striatum are attributable to the dopaminer- cassettes for 2 ([H]SCH23390) or 4 weeks ([3H])spiro-
gic nigrostrial axons or to intrinsic striatal neurons, peridol), and developed in Kodak D19 at 18C.
and second, to identify the cellular associations of the
two receptor subtypes in the globus pallidus, ento- Dala analysis
peduncular nucleus and substantia nigra, the three Analysis of the autoradiograms was accomplished using
targets of striatifugal axons. a Wild M420 macroscope with a stabilized light source
(Stahl Research Labs) and a Dage-MT1 series 68 video
camera. The video signal is digitized by the PCVision-plus
frame grabber (Imaging Technology) and analysed with
EXPERIMENTAL PROCEDURES software developed by R.M.B. and M. Wong. This system
is linearly related to and calibrated against transmittance
Lesions values obtained with a Leitz MPV compact photometer.
Adult, domestic cats of either sex were deeply anesthe- First. a standard curve was generated from calibrated
tized for surgery with halothane and injected with either tritium microscales relating gray value to the tissue wet-
ibotenic acid (Sigma) at a concentration of IOpg/pl of weight equivalent concentration of tritium. Since the linear
0.1 M NaPO, buffer (pH 7.4), or 6-hydroxydopamine response range for optical density of the LKB Ultrofilm lies
(Sigma) at a concentration of 25 pg/pl of 0.1% ascorbate- between approximately 0.08 and 0.8, only measurements in
saline (pH 7.0). The neurotoxin solutions were ejected from this range were used. Next the gray level measurements from
a 5~1 Hamilton syringe attached to the Kopf stereotaxic various areas of the total and non-specific images were
apparatus. The tip of the needle was positioned according interpolated on the standard curve by linear regression
to stereotaxic coordinates selected from the atlas of Jasper analysis and converted to fmol/mg of tissue wet-weight
and Ajmone-Marsan,*9 and the neurotoxins were ejected at by dividing the interpolated value by a constant equal to
a volume of 0.5 ~1 and a rate of 0.05 pl/min. The needle was 1000 divided by the specific activity (in Ci/mmol) of the
left in place for 10 min following completion of each injec- incubation solution.
tion. For ibotenate-induced, axon-sparing lesions that were The values obtained from measurements of the non-lesion
confined to the striatum,25 one to five separate penetrations side did not differ significantly from measurements made in
were made at different coordinates to produce lesions in several naive cats.8 Therefore, measurements of specific
either the caudate nucleus (n = 4) or the putamen (n = 2). binding in regions within basal ganglia structures on the
For nigral lesions (n = 8) the cats were injected sub- lesion side were compared as closely as possible to the
cutaneously with desmethyhmipramine (I5 mg/kg in saline) corresponding regions on the non-lesion side in each case,
40min prior to surgery. Three penetrations were made at and the changes in binding are expressed with respect to the
different coordinates in an attempt to maximize coverage of non-lesion side. In each case, the caudate nuclei were
 Dopamine receptor associations 853

measured at 10 rostocaudal levels, the putamen at six, the
globus pallidus, entopeduncular nucleus, and pars reticulata
of the substantia nigra at four levels each, and the
pars compacta at two rostrocaudal levels. The lesion and
non-lesion values for each structure were compared using
Students t-test for unpaired data. Differences were con-
sidered significant at P < 0.05.
RESULTS
The eficts of ibotenate lesions of the striatum
Nissl-stained tissue sections through the area of the
lesions revealed areas of severe neuronal destruction
and extensive gliosis (Fig. 1). Within the rather
clearly demarcated borders of the infusion zones,
there invariably remained a small number of neuron-
like cells. The sparing of some striatal cells within the
lesion zone was to be expected from previous litera-
ture concerning excitototoxin-induced lesions of the
striatum.36 Associated with the area of cell loss was
a dramatic reduction in the binding of both D, and
D, receptor radioligands as compared to the intact
side (Fig. 2A, B). The binding concentrations of
either radioligand on the non-lesion side were not
significantly different from naive animals. As in the
N&l-stained image, the border of the lesion is clearly
visible and abrupt in the autoradiograms (Figs 3A
and 4A). Within the damaged area, the reduction
in [3H]SCH23390-binding was nearly complete,

ac nucleus accumbens ot optic tract

corpus callosum PU putamen
2 caudate nucleus rr retrorubral area
cg central gray substance S septum
cp cerebral peduncle snc substantia nigra pars compacta
ep entopeduncular nucleus sni substantia nigra pars lateralis
giobus pallidus snr substantia nigra pars reticulata
? internal capsule sth subthalamic nucleus
iP interpeduncular nucleus V lateral cerebral ventricle
ml medial lemniscus

Fig, 1. A photomicrograph of a Cresyl Violet-stained tissue section through the dorsomedid quadrant
of the head of the caudate nucleus showing part of the area involved by the ibotenate infusion. The area
of severe neuron loss and gliosis can be seen to the left; its border with relatively undamaged tissue along
the lateral ventricle is marked by the series of black arrows. The open arrows indicate the position of two
infusion sites. Scale bar = 1mm.
 854 R. M. BECKSTEAD

averaging 94%, whereas the reduction of of the nigra after large lesions of the caudate nucleus.
[Hlspiroperidol-binding was somewhat less severe, This reduction was smaller (31%) than that in the
averaging around 85%. Beyond the borders of dimin- pars reticulata, but was significant.
ished binding, the values of both the D, and D, The binding of [3H]spiroperidol was reduced by
receptor radiohgands were comparable to the non- about half in the topographically related parts of the
lesion side. Finally, the patchy patterns of high- and globus pallidus (55%; Fig. 2B) and entopeduncular
low-density radiohgand-binding which are displayed nucleus (53%) after caudate and putamen lesions.
on the intact side were eradicated within the area of However, no statistically significant differences be-
ibotenate infusion (Figs 3A and 4A). tween the lesion side and the intact side could be
The ibotenate-induced lesions of the striatum also detected in either the pars compacta or pars reticulata
caused reductions of D, and D, receptor-binding in of the substantia nigra (Fig. 2B).
the ipsilateral pallidal and nigral targets of striatal
The effects of Ghydroxydopamine lesions of nigral
axons (Fig. 2). These reductions were typically non-
dopamine neurons
uniform within the target nuclei-although large
lesions caused a widespread reduction of radioligand- Six cats were subjected to unilateral lesions of the
binding, a particular zone of excessive depletion was substantia nigra dopaminergic neuron population
usually evident which presumably corresponds to the with 6-hydroxydopamine. This procedure resulted in
territory that would normally receive axons from the a severe loss of neurons and an extensive glial re-
damaged part of the striatum (Figs 3B-E and 4B-D). sponse in the pars compacta of the substantia nigra
Since no D, receptors are measurable in the globus (Fig. 5). Associated with this cell loss was a
palhdus of intact cats, it follows that there could be significant, 95% reduction of [3H]spiroperidol-
no reduction in the lesion cases. However, the density binding in the area typically dominated by pars
of [3H]SCH23390-binding in those parts of the ento- compacta dopaminergic neurons (Fig. 6B). This
peduncular nucleus and pars reticulata of the sub- diminution of D, receptor-binding can be seen clearly
stantia nigra topographically related to the striatal in the autoradiograms when the lesion side is
lesion zone diminished by 75% and 81%, respectively compared to the non-lesion side (Fig. 7A, B). There
(Fig. 2A). A diffuse diminution of D, receptor- was also a smaller (39%), but significant loss of D,
binding was detected also in the pars compacta region receptor-binding in the area of the pars reticulata
(Fig. 6A). The binding of [H]SCH23390 was also
significantly reduced by 17% in the area of the pars

A
compacta, but not in the area of the pars reticulata

*
3 of the substantia nigra (Fig. 6A).
+ ml-
E Quantitative autoradiography of basal ganglia

*
2 structures on the two sides of the brain in all
E.200 - 6-hydroxydopamine cases failed to reveal any

Lbb
$ significant differences between the lesion and non-
w 100
lesion sides in the overall amounts of D, or Dz
l radioligand bound in individual striatal or pallidal
$ *
P structures (Fig. 6A, B). Moreover, with the exception
0
0 _L of the nigral subnuclei on the lesion side, the binding
CD/PU GP E!J SNr SNC
of either radioligand did not differ significantly from
naive animals in either amount or distribution pat-
B terns within the basal ganglia. Finally, no lesion-
induced changes could be detected in the hetero-
geneous density patterns within the caudate nucleus
or putamen between the lesion and non-lesion sides
(Fig. 8A, B).

DISCUSSION

GP EP SNr Lesion-induced changes in the striatum

basal ganglia structure
Ibotenate infusions invariably produced reductions
Fig. 2. Comparison of the mean values (N = 6) of specific in the amount of D, and D, receptor concentrations
D, (A) and D, (B) radioligand binding in various basal within the area of striatal cell loss as compared to the
ganglia components on the non-lesion (black bars) and non-lesion side (94% for D, and 85% for D2). The
lesion (white bars) sides after an ibotenate infusion in the magnitudes of these reductions are comparable to
caudate nucleus on one side. The error bars equal one
results obtained with neurotoxin-induced lesions in
standard error of the mean. Students r-test for unpaired
data was used to estimate the significance of the differences the rat striatum by others,~3~2.28~3 and seem to be a
(*P < 0.005; **p < 0.05). direct consequence of removing the cells that syn-
 Dopamine receptor associations 855

Fig. 3. Film autoradiograms of basal ganglia structures to compare [jH]SCH23390 (D,) binding on the
lesion (left) and non-lesion (right) sides. The faint area in the caudate nucleus in (A) indicates the extent
of reduced binding due to the ibotenate infusion (x I). (B) and (C) show the entopeduncular nuclei, and
(D) and (E) show the substantia nigras at x 6.3 magnification. Scale bars = 1 mm.

thesize the receptors rather than a secondary effect on
a system influenced by the striatum. The persistence
of some specific receptor-binding within the lesion
zone may be a consequence of (1) the survival of a
few striatal neurons that express the receptors, (2)
receptors in processes from striatal cell bodies lying
outside the lesion zone, (3) the presence of dopamine
receptors in the spared striatal afferent axons and (or)
ghal cells, or (4) some combination of these factors.
Regarding the residual receptors of the Dz subtype,
their presence in afferent axons from the substantia
nigra seems to be a possibility since the existence of
such presynaptic D, receptors has already been sug-
gested in the rat. 12.49Cerebral cortical axons in the
856 R. M. BECKSTEAD

Fig. 4. Film autoradiograms of basal ganglia structures to compare [3H]spiroperidol (DJ binding on the
lesion (left) and non-lesion (right) sides. The faint area in the caudate nucleus in (A) indicates the extent
of reduced binding due to the ibotenate infusion ( x 1). Binding in the globus palhdus on the lesion and
non-lesion sides is shown in (B) and (C), respectively ( x 6.3). The ventral portion of the midbrain including
the substantia nigras is shown in (D) at x 4 magnification. Scale bars = 1mm.
 Dopamine receptor associations 857

Fig. 5. Photomicrographs of Cresyl Violet-stained tissue sections at a caudal level of the ventral midbrain
showing the middle mediolateral portions of the substantia nigra of either side. Note the extensive neuron
loss and gliosis on the side of the injection of 6-hydroxydopamine (A). The corresponding region of the
non-lesion side is shown in (B) for comparison. Medial is toward the border of the two images. The arrow
in (A) indicates a glial scar presumably related to the tip of the infusion cannula. Scale bar = 1 mm.

rats striatum have also been reported to contain D,
The presence of such receptors in
receptors. 22*24S3**54J8
corticostriatal axons of the cat may account for some
of the D2 preservation in the present cases. However,
dopamine receptors have not been demonstrated in
cat corticostriatal axons, and such a demonstration
now seems improbable in light of the recent, well-
controlled studies which deny the existence of D,
receptors in the rats corticostriatal axons3. In any
case, the present results indicate clearly that the vast
majority of both D, and D, receptors in the cat
striatum are located on intrinsic striatal neurons.
Other than the changes within the substantia nigra,
the 6-hydroxydopamine-induced destruction of do-
pamine neurons in the cat failed to alter the binding
of either the D, or D2 radioligand in striatal and
pallidal structures with respect to the non-lesion side.
The same observation has been made recently in the
rat brain.g*21Filloux ef al.* surmised that the absence
of change was a consequence of a reduction in
presynaptic D2 receptors which was balanced by a
proportional up-regulation of postsynaptic D2 recep
tors. Other authors, however, have successfully de-
tected a 2040% increase in D, receptor-binding to
 R. M. BECKSTEAD
A striatonigral D, receptors. Even within the pallidal
CDiPU
B tors in the pallidum and nigra are associated with
CDIPU W EP SNr SNC
basal ganglia structure
Fig. 6. Comparison of the mean values (N = 8) of specific
D, (A) and Dz (B) radioligand binding in various basal
ganglia components on the non-lesion (black bars) and
lesion (white bars) sides after a Ghydroxydopamine infusion
in the region of the pars compacta on one side. The error
bars are one standard error of the mean. Students l-test for
unpaired data was used to estimate the significance of the
differences (*P < 0.005; **P cc 0.05).
homogenates and tissue sections of the dopamine
denervated striata of rats.4*s~44.53Increased
[3H]spiroperidol-binding density has also been re-
ported in autoradiograms of the striatum of
l-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated
monkey. 33 Other than the present experiments, no
studies have been performed to date in the cat brain
on dopamine denervation-induced changes of recep-
tor density in the striatum.
Perhaps more importantly, the 6-hydroxy-
dopamine-indu~d destruction of nigral dopamine
neurons had no effect on the patterns of high- and
low-density radioligand-binding in the cats caudate
nucleus. Coupled with the observation that the
heterogeneous patterns are completely eradicated by
striatal ibotenate lesions, this finding indicates that
the high- and low-density zones are determined by the
relative densities of receptors in striatal cell popu-
lations and not by nigrostriatal axons.
Lesion-induced changes in pallidal und nigral ceil
groups
The changes in the pallidum and nigra caused by
the subtotal, striatal ibotenate lesions appeared to
reflect the topographic nature of the striatonigral
projection in that the depletions were not uniform,
but instead exhibited regions of relatively pro-
nounced receptor loss and regions where there was
only slight diminution. This phenomenon was ob-
served earlier by Altar and Hauser for the rat
and nigral portions showing the greatest depletion,
however, the receptor-binding was not completely
eliminated. Instead, the maximum depletions were
approximately 75% for D, and 50% for D, receptors.
Assuming the depletion was a result of striatal axon
loss, this finding indicates that the topographic or-
ganization in the striatopallidal and -nigral
projections may be somewhat less than strict. An-
other possibility is that some of the dopamine recep-
tissue elements other than the striatal axons such as
the intrinsic neurons or non-striatal afferent axons.
Only direct, axon-sparing lesions of the pallidal and
nigral components themselves can answer this ques-
tion definitively.
After unilateral, 6-hydroxydopamine-indu~d le-
sions of the pars compacta ceils, the major difference
between the two sides occurs within the substantia
nigra itself where the D, receptor density is reduced
by 93% in the area of the pars compacta and 40%
in the area of the pars reticulata on the side of the
6-hydroxydopamine-induced lesion. This finding in
the cat is not surprising in light of the already
substantial evidence from lesion experiments in the
rat which shows that the D, receptors in the substan-
tia nigra are located mainly on dopamine neurons of
the pars compacta. 21,38,46 The present observation of
a signi~~ant reduction of [3H]spiro~ridol-binding in
the pars reticula& seems likely to result from loss of
D, receptor-containing dendrites of pars compacta
neurons. That this depletion of Dz receptors in the
pars reticulata was less than half indicates that many
of the D, receptors are associated with non-
dopaminergic tissue elements. Somewhat sur-
prisingly, the remaining D, receptors cannot be as-
signed to striatonigral axons since the striatal lesions
did not result in reduced [3H]spiroperido1-binding in
the pars reticulata. The findings of Ruffieux and
SchultzSo are pertinent to this point. They found that
pars reticulata neurons which project to the thalamus
are responsive to iontophoretically applied do-
pamine, thus suggesting the presence of dopamine
receptors on such cells. More recently, Heyer et a1.17
found that ventral mesencephalic neurons grown in
culture express D, receptors, as assessed by the
binding of [Hlspiroperidol, even after the dopami-
nergic population has been eliminated by
6-hydroxydopamine toxicity. These two obser-
vations, together with the present binding data, raise
the possibility that the non-dopamine cells of the pars
reticulata express D2 receptors as do the dopamine
cells of the pars compacta. Further work is required
to definitively answer this important question.
Earlier studies on the rat nigra have disassociated
D, receptors and dopamine neurons; after
6-hydroxydopamine-induced lesions, the dopamine
stimulation of adenylate cyclase reportedly remained
intact?.46,5 Conversely, excitotoxin-induced lesions
 Dopamine receptor associations 859

Fig. 7. Autoradiograms of tissue sections through the substantia nigras to compare radioligand binding
on the lesion (A and C) side to the non-lesion (I3 and D) side. The D, radioligand binding is depicted
in (A) and (B), and the D, radioligand binding in (C) and (D). Note the reduced binding of
13H]spiroperidolin the pars compacta on the lesion side (A). The midline is toward the center of the figure.
Scale bar = I mm.

of the rat caudatoputamen resulted in decreased
binding of the D, radioligand rHlSKF38393 in the
pars reticulata,3,9 and various interruptions of the
striatonigral projection caused a massive reduction in
dopamine-stimulated, D,-dependent adenylate cy-
clase activity. 23.M*55
Likewise, D,-mediated regulation
of the phosphoprotein DARPP-32 is greatly reduced
by a striatal lesion, but not by a lesion of the pars
compacta neurons. 37 Therefore, the preservation of
these enzyme activities was attributed to the stimu-
lation of D, receptors in striatal axons rather than
dopamine neurons. Two recent reports contradict
this finding. First, Porceddu et ~1.~ found a 60%
reduction of dopamine-sensitive adenylate cyclase
activity which is accompanied by a 27% reduction in
[3H]SCH23390-binding to nigral homogenates from
rat brains 15 days after unilateral infusions of
6-hydroxydopamine in the medial forebrain bundle.
Second, Savasta et al. used an autoradiographic
technique in the rat to show that
[3H]SCH23390-binding is reduced in the pars com-
pacta, but not in the pars reticulata, by
6-hydroxydopamine-induced lesions of the dopamine
neurons. On the basis of their respective obser-
vations, both groups concluded that dopamine neu-
rons of the pars compacta do express receptors of the
D, as well as the DZ subtype. An alternative expla-
nation is that the apparent reduction of D,-binding in
the area of the pars compacta is a secondary effect of
removing the targets of D,-containing striatonigral
axons and a subsequent reorganization of such
axons. In an attempt to address this possibility,
Savasta et al. demonstrated that ibotenate-induced
lesions of the caudatoputamen do not cause a reduc-
tion of [3H]SCH23390-binding in the pars compacta,
but only in the pars reticulata. Superficially, this
finding would seem to support the conclusion that the
D, receptors of the pars compacta are not assignable
to the striatonigral axons. However, since the striat-
onigral projection to the pars compacta originates in
860 R. M. BECKSTEAD

Fig. 8. Autoradiograms of adjacent frontal sections through the head of the caudate nucleus to show (A)
D, and (B) D, radioligand binding in the striatum 6 weeks after a unilateral infusion of
6-hydroxydopamine in the vicinity of the substantia nigra. The lesion side is on the viewers right.
 Dopamine receptor associations 861

the ventral caudatoputamen and nucleus accu- the rat.10J6J7*52 We made the additional discovery of
mbens,39 the lesions produced by Savasta er al.) an unprecedented relationship between the Dl and D,
would not involve the striatal axons to the pars receptor patterns in the caudate nucleus of the cat;
compacta. namely, that the high-density zones of Di receptors
The present data on the cats nigra are entirely are sometimes in precise register with the low-density
consistent with the idea that most of the D, receptors zones of D, receptors in the caudate nucleus.8 The
in the pars compacta, like those in the pars reticulata,
present report furthers our knowledge of dopamine
are associated with striatal axons; the small reduc- receptors in the cat basal ganglia by implicating the
tions that occur in specific [H]SCH23390-binding tissue elements responsible for these selective distri-
after 6-hydroxydopamine lesions do not reach statis- butions of the two dopamine receptor subtypes.
tical significance and more likely seem to be a con- Over the past decade, there has been an explosion
sequence of non-specific (mechanical) damage to a of research on the basal ganglia that has led to some
few of the striatonigral fibers at the infusion site important new concepts of corpus striatal or-
rather than the loss of dopamine neurons. If the ganization. Foremost among these new concepts is
majority of D, receptors in the pars compacta are the notion that the striatum, or input side of the
located on striatal axons, it seems likely that they corpus striatum, is organized into compartments that
would arise from cells of the nucleus accumbens and can be identified by a number of cytochemical,
ventral caudate nucleus since these striatal areas are connectional, and physiological criteria. The present
known to project selectively to the pars compacta in observation that the heterogeneous pattern of D, and
the rat. Thus, lesions of the nucleus accumbens and D, receptors in the striatum is a product of intrinsic
ventral caudate nucleus might be expected to produce neurons lends further support to the notion of a
a reduction in D, receptor-binding in the area of the functional compartmentation of striatal cell popu-
pars compacta even larger than the 30% seen in the lations that is likely to be important for the basal
present cases of caudate nucleus lesion. However, ganglias role in behavioral control. The present
since none of the subtotal striatal lesions of the finding that the differential distributions of the two
present study eliminate [3H]SCH23390-binding en- dopamine receptor subtypes in pallidal and nigral
tirely, and no lesions of the nucleus accumbens have structures is mainly attributable to axons of striatal
been produced in this or any other published study origin reinforces the concept of a duality in the
to date, it remains possible that a fraction of the D, striatopallidal-nigral projection system. Moreover,
receptors are associated with dopamine neurons of this same observation strongly suggests that striatal
the pars compacta. cell compartments which send axons to the globus
pallidus (external segment) either do not express D,
receptors or do not transport such receptors ortho-
CONCLUDING REMARKS
gradely in their axons.) Although there is now
evidence that some striatal cells express both receptor
Recently my own lab and that of Richfield et ~1.~
subtypes,42 it seems reasonable to posit the existence
reported that dopamine receptors of both the D, and
of a population of striatal cells which sends axons
D2 subtype are differentially distributed in the cats
selectively to the globus pallidus,5*20,4 and does not
basal ganglia in at least two important ways. First,
synthesize substance P6*26or D, dopamine receptors.
they are distributed in irregular high- and low-density
An important course in future basal ganglia research
zones in the striatum, a situation that has been
will be to elucidate how these compartmentalized
reported for D2 receptors in the monkey and human
mechanisms interact to produce an operational net-
striatum as well.32*35Second, there is a differential
work.
distribution of the two dopamine receptor subtypes in
the pallidal and nigral targets of striatal axons,*.48 Acknowledgements-This work was supported by NSF
which is similar to that reported in the monkey and grant BNS 8504438. I thank Ms. T. Pickens for expert
human,* but more exclusive than that reported for technical assistance.

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(Accepted 11 May 1988)