A horseradish peroxidase study of afferent connections of the globus pallidus in Macaca mulatta

Summary The high tonic discharge rates of globus pallidus neurons in awake monkeys suggest that these neurons may receive some potent excitatory input. Because most current electrophysiological evidence suggests that the major described pallidal afferent systems from the neostriatum are primarily inhibitory, we used retrograde transport of horseradish peroxidase (HRP) to identify possible additional sources of pallidal afferent fibers. The appropriate location was determined before HRP injection by mapping the characteristic high frequency discharge of single pallidal units in awake animals. In animals with injections confined to the internal pallidal segment, retrograde label was seen in neurons of the pedunculopontine nucleus, dorsal raphe nucleus, substantia nigra, caudate, putamen, subthalamic nucleus, parafascicular nucleus, zona incerta, medial and lateral subthalamic tegmentum, parabrachial nuclei, and locus coeruleus. An injection involving the external pallidal segment and the putamen as well resulted in additional labeling of cells in centromedian nucleus, pulvinar, and the ventromedial thalamus.Abbreviations AC anterior commissure - CG central grey - CM centromedian nucleus - CN caudate nucleus - DM dorsomedial nucleus - DR dorsal raphe nucleus - DSCP decussation of superior cerebellar peduncle - GPe globus pallidus, external segment - GPi globus pallidus, internal segment - LC locus coeruleus - LL lateral lemniscus - MG medial geniculate nucleus - ML medial lemniscus - NVI abducens nucleus - OT optic tract - Pbl lateral parabrachial nucleus - Pbm medial parabrachial nucleus - Pf parafascicular nucleus - PPN pedunculopontine nucleus - PuO oral pulvinar nucleus - RN red nucleus - SCP superior cerebellar peduncle - SI substantia innominata - SNc substantia nigra, pars compacta - SNr substantia nigra, pars reticulata - STN subthalamic nucleus - TMT mamillothalamic tract - VA ventral anterior nucleus - VLc ventral lateral nucleus, pars caudalis - VLm ventral lateral nucleus, pars medialis - VLo ventral lateral nucleus, pars oralis - VPI ventral posterior inferior nucleus - VPM ventral posterior medial nucleus - VPLc ventral posterior lateral nucleus, pars caudalis - ZI zona incerta     

Pallidotectal projection to the inferior colliculus of the rat

Summary After injection of fluorescent tracer into the inferior colliculus (IC), retrogradely labeled cells were observed not only in the temporoauditory cortex (ACx) and the substantia nigra pars lateralis, but also in the globus pallidus (GP). These labeled GP cells were localized exclusively in the caudal portion of the GP, which has been known to project to the ACx. Employing a retrograde fluorescent double labeling technique, the GP-IC neurons were found to be distributed in a separate manner from the GP-ACx neurons within the caudal GP. The present study provides further anatomical evidence that the caudal GP has a functional role in auditory processing.Abbreviations ACx temporoauditory cortex - BC Brachium conjunctivum - CP cerebral peduncle - CPu caudate putamen - DY Diamidino Yellow - EP entopeduncular nucleus - FG Fluoro-Gold - GP globus pallidus - I internal capsule - IC inferior colliculus - OT optic tract - SC superior colliculus - SN1 substantia nigra pars lateralis - T thalamus - TB True Blue - TPC nucleus tegmenti pedunculopontinus pars compacta     

Topographic projections from the basal ganglia to the nucleus tegmenti pedunculopontinus pars compacta of the cat with special reference to pallidal projections

Summary Projections from the basal ganglia to the nucleus tegmenti pedunculopontinus pars compacta (TPC) were studied by using anterograde and retrograde tracing techniques with horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) in the cat. Following WGA-HRP injections into the medial TPC area, a substantial number of retrogradely labeled cells were seen in the entopeduncular nucleus (EP) and medial half of the substantia nigra pars reticulata (SNr), whereas following WGA-HRP injections into the lateral TPC area, labeled cells were marked in the caudal half of the globus pallidus (GP) and lateral half of the SNr. To confirm the retrograde tracing study, WGA-HRP was injected into the EP or the caudal GP, and anterograde labeling was observed in the TPC areas. Terminal labeling was located in the medail TPC area in the EP injection case, while terminal labeling was observed in the lateral TPC area in the caudal GP injection case. Projections from the striatum to the pallidal complex (the EP and the caudal GP) were also studied autoradiographically by injecting amino acids into various parts of the caudate nucleus and the putamen. Terminal labeling was distributed over the whole extent of the EP and the rostral GP following injections into the rostral striatum (the head of the caudate nucleus or the rostral part of the putamen), while terminal labeling was distributed over the caudal GP following injections into the caudal striatum (the body of the caudate nucleus or the caudal part of the putamen). From these findings, we conclude that there exists a medio-lateral topography in the projection from the basal ganglia to the TPC: The EP receives afferent projections from the rostral striatum and projects to the medial TPC area, whereas the caudal GP receives projections from the caudal striatum and sends fibers to the lateral TPC area.Abbreviations BC brachium conjunctivum - CD caudate nucleus - CP cerebral peduncle - DBC decussation of the brachium conjunctivum - EP entopeduncular nucleus - GP globus pallidus - IC internal capsule - ICo inferior colliculus - LH lateral habenular nucleus - ML medial lemniscus - PN pontine nuclei - PUT putamen - SCo superior colliculus - SI substantia innominata - SN substantia nigra - SNc substantia nigra pars compacta - SNr substantia nigra pars reticulata - STN subthalamic nucleus - TH thalamus - TPC nucleus tegmenti pedunculopontinus pars compacta     

An autoradiographic study of efferent connections of the globus pallidus in Macaca mulatta

Summary Radioactive amino acids were injected into restricted regions of the globus pallidus of rhesus macaques to allow identification of the organization and courses of efferent pallidal projections. The previously identified projection of the internal pallidal segment (GPi) to ventral thalamic nuclei showed a topographic organization, with the predominant projection from ventral GPi being to medial and caudal ventralis anterior (VA) and lateralis (VL) and from dorsal GPi to lateral and rostral VA and VL. Pallidal efferent fibers also extended caudally and dorsally into pars caudalis of VL, but they spared the portion of pars oralis of VL shown by others to receive input from the cerebellum. In addition to centromedian labeling in all animals, the parafascicular nucleus was also labeled when isotope was injected into dorsal GPi. The medial route from GPi to the midbrain tegmentum was more substantial than has been shown before, and along this route there was an indication that some fibers terminated in the prerubral region. The projection to the pedunculopontine nucleus was extensive, and fibers continued caudally into the parabrachial nuclei.Pallidal projections to the thalamus seem to be topographically organized but spare thalamic regions that interact with area 4. Caudally directed efferent fibers follow multiple routes and extend more caudally than to the pedunculopontine nuclei.Abbreviations Cd caudate nucleus - CM centromedian nucleus - CT central tegmental tract - DPCS decussation of superior cerebellar peduncle - F fornix - FLM medial longitudinal fasciculus - GPe globus pallidus, pars externa - GPi globus pallidus, pars interna - HbL lateral habenular nucleus - HbM medial habenular nucleus - Is interstitial nucleus - LM medial lemniscus - MD dorsomedial nucleus - PbL lateral parabrachial nucleus - PbM medial parabrachial nucleus - PCS superior cerebellar peduncle - Pf parafascicular nucleus - PPN pedunculopontine nucleus - Put putamen - R reticular nucleus - Rmg red nucleus, pars magnocellularis - Rpc red nucleus, pars parvocellularis - S stria medullaris - SI substantia innominata - SNc substantia nigra, pars compacta - SNr substantia nigra, pars reticulata - St subthalamic nucleus - ST stria terminalis - THI habenulointerpeduncular tract - TM tuberomamillary nucleus - TMT mamillothalamic tract - VA nucleus ventralis anterior - VAmg nucleus ventralis anterior, pars magnocellularis - VAp nucleus ventralis anterior, pars principalis - VI nucleus ventralis intermedius - VLc nucleus ventralis lateralis, pars caudalis - VLm nucleus ventralis lateralis, pars medialis - VLo nucleus ventralis lateralis, pars oralis - VPL nucleus ventralis posterior lateralis - X area X Supported by National Institutes of Health, grant RR00166, Rehabilitation Services Administration, grant 16-P-56818, and PHS grant NS10804     

The subcortical afferents to caudate nucleus and putamen in primate: A fluorescence retrograde double labeling study

The cellular origin and degree of collateralization of the subcortical afferents to the caudate nucleus and the putamen in squirrel monkeys (Saimiri sciureus) were studied using the following combinations of fluorescent retrograde tracers: Evans blue and DAPI-Primuline, Fast blue and Nuclear yellow, True blue and Nuclear yellow. After the injections, cells containing the tracer delivered in caudate nucleus (caudate-labeled cells) and others labeled with the complementary tracer injected in putamen (putamen-labeled cells) occur in large number in intralaminar nuclei, substantia nigra pars compacta, midbrain raphe nuclei and central midbrain tegmentum. In addition, a small to moderate number of putamen-labeled cells is found in external pallidum, pulvinar and laterodorsal thalamic nuclei, and basolateral amygdaloid nucleus, whereas some caudate and putamen-labeled cells are scattered in ventral tegmental area and locus coeruleus. However, very few double-labeled cells are present in all these structures. In rostral intralaminar nuclei, the labeled cells are not confined to the known cytoarchitectonic boundaries of the nuclei but impinge slightly upon ventrolateral and mediodorsal nuclei. At this level, the caudate-labeled cells lie more dorsally and medially relative to putamen-labeled cells, but a high degree of intermingling exists and some double-labeled cells occur particularly in nucleus centralis lateralis. In caudal intralaminar nuclei, caudate-labeled cells are strictly confined to parafascicular nucleus and putamen-labeled cells present only in centre median, without any overlap between the two neuronal populations. In substantia nigra pars compacta, clusters of caudate-labeled cells are closely intermingled with clusters of putamen-labeled cells according to a complex mosaic-like pattern that varies along the rostrocaudal extent of the structure. Overall, however, caudate-labeled cells predominate rostrodorsally and putamen-labeled cells are more abundant caudoventrally in substantia nigra pars compacta, with only a few double-labeled cells. Some caudate and putamen-labeled cells are also scattered in contralateral substantia nigra pars compacta. In dorsal raphe nucleus, putamen-labeled cells tend to occupy a more lateral position relative to caudate-labeled cells, with again very few double-labeled neurons. The caudate and putamen-labeled cells are less numerous and more closely intermingled in nucleus centralis superior. Numerous striatal afferent cells are also found bilaterally in the peribrachial region of midbrain tegmentum, comprising the pedunculopontine nucleus area. There, the putamen-labeled cells are slightly more numerous than the caudate-labeled cells with less than 10% of these neurons being double-labeled.

Our findings suggest that the subcortical afferents to caudate and to putamen in primates arise largely from different neurons in thalamus and midbrain. These two types of striatal afferent neurons are distributed according to various patterns that are much more complex than could have been inferred from current knowledge of the topographical organization of striatal afferents.     

Striatal substance P cell clusters coincide with the high density terminal zones of the discontinuous nigrostriatal dopaminergic projection system in the cat: a study by combined immunohistochemistry and autoradiographic axon-tracing

A portion of the nigrostriatal projection that originates from presumably dopaminergic neurons in the caudal pars compacta of the substantia nigra and the suprajacent pars dorsalis (retrorubral area), was shown by [3H]amino acid autoradiographic tracing to distribute nonhomogeneously in the head of the caudate nucleus, such that zones of high density termination are in register with the archipelago of substance P cell clusters revealed immunohistochemically in the same and adjacent tissue sections of the cat's brain. Axons from this same portion of the substantia nigra distribute densely at caudal levels of the putamen where again substance P-immunoreactive striatal cells are numerous. In nearby tissue sections from the same cases, tyrosine hydroxylase-like immunoreactivity suggested only subtle variations in the density of the catecholamine axon network within the striatum. Thus, whereas dopamine axons are distributed densely throughout the striatum, those originating from cells in the caudal pars compacta et dorsalis of the substantia nigra and ending in the head of the caudate nucleus appear to terminate preferentially within the substance P cell clusters. These data suggest that the striatal substance P cells, which send their axons selectively to the entopeduncular nucleus and substantia nigra, but much less so the globus pallidus, are a major target of nigrostriatal dopamine transmission. This result is discussed with respect to the anatomical, neurochemical and functional organization of the striatifugal projection system.     

Metabolic activity of excitatory parafascicular and pedunculopontine inputs to the subthalamic nucleus in a rat model of Parkinson's disease

Using a combination of metabolic measurement and retrograde tracing, we show that the neurons in the pedunculopontine nucleus and parafascicular nucleus of the thalamus that project to the subthalamic nucleus are hyperactive after nigrostriatal dopaminergic denervation in rats. In Parkinson's disease, the loss of dopaminergic neurons induces a cascade of functional changes in the basal ganglia circuitry including a hyperactivity of the subthalamic nucleus. This hyperactivity is thought to be due to a diminution of the inhibitory pallidal influence. However, recent studies have suggested that other cerebral structures are involved in the subthalamic neuronal hyperactivity. This study was undertaken to identify these cerebral structures. Neurons projecting to the subthalamic nucleus were identified by retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase, injected into the subthalamic nucleus of rats with 6-hydroxydopamine unilateral lesion of the substantia nigra pars compacta and sham-lesioned animals. Metabolic activity was determined in the same neurons using in situ hybridization for the first subunit of cytochrome oxidase messenger RNA, a metabolic marker, and image analysis. Horseradish peroxidase-labeled neurons were found in the globus pallidus, parafascicular and pedunculopontine nucleus and sometimes in raphe nuclei and the substantia nigra pars compacta. Measurement of metabolic activity was performed for the globus pallidus, the pedunculopontine and parafascicular nuclei. The expression level of the first subunit of cytochrome oxidase messenger RNA in neurons projecting to the subthalamic nucleus was 62% higher in parafascicular neurons and 123% higher in pedunculopontine neurons in 6-hydroxydopamine-lesioned rats, compared to sham-lesioned animals. An increase was also observed in the globus pallidus, but did not reach significance.Our results suggest that hyperactivity of subthalamic neurons could be due, at least in part, to an increase of excitatory input arising from the pedunculopontine and parafascicular nuclei. These data also suggest that the latter structures may play an important role in the physiopathology of Parkinson's disease.     

Direct projections from the central amygdaloid nucleus to the globus pallidus and substantia nigra in the cat.

Employing both anterograde and retrograde axonal tracing, we investigated direct projections from the central amygdaloid nucleus to the basal ganglia in the cat. The anterograde axonal tracing of Phaseolus vulgaris-leucoagglutinin revealed that projection fibers from the central amygdaloid nucleus to the basal ganglia ended in the globus pallidus (the feline homolog to the external segment of the globus pallidus of primates) and substantia nigra. The amygdalopallidal fibers terminated chiefly in the medial most part of the globus pallidus at its caudal level. The amygdalonigral fibers terminated densely in the substantia nigra pars lateralis, and moderately in the dorsolateral part of the substantia nigra pars reticulata; none of them were found to end in the substantia nigra pars compacta. Both of the amygdalopallidal and amygdalonigral projections were ipsilateral. These neuronal connections were confirmed by retrograde axonal tracing of cholera toxin B subunit in the second set of the experiments: The cells of origin of the amygdalopallidal and amygdalonigral projections were located predominantly in the lateral part of the central amygdaloid nucleus, and additionally in the intercalated cell islands of the amygdala. Most of them were of small bipolar or multipolar type. The cells projecting to the globus pallidus were preferentially distributed at the rostral levels of the central nucleus and intercalated cell islands of the amygdaloid complex, while those projecting to the substantia nigra were mainly located at the caudal levels of these amygdaloid subdivisions. In the third set of the experiments, sequential double-antigen immunofluorescence histochemistry for transported cholera toxin B subunit and horseradish peroxidase showed that some single neurons in the lateral part of the central amygdaloid nucleus, particularly at its middle level, issued axon collaterals to both the globus pallidus and substantia nigra pars lateralis. The results of the present study indicate that the central amygdaloid nucleus sends projection fibers to the globus pallidus and substantia nigra possibly to exert a limbic influence upon forebrain motor mechanisms.     

Distribution of divalent metal transporter-1 in the monkey basal ganglia

An accumulation of iron occurs in the brain with age, and it is thought that this may contribute to the pathology of certain neurodegenerative diseases, including Parkinson's disease. In this study, we elucidated the distribution of divalent metal transporter-1 (DMT1) in the monkey basal ganglia by immunocytochemistry, and compared it with the distribution of ferrous iron in these nuclei by Turnbull's Blue histochemical staining. We observed a general correlation between levels of DMT1, and iron staining. Thus, regions such as the caudate nucleus, putamen, and substantia nigra pars reticulata contained dense staining of DMT1 in astrocytic processes, and were also observed to contain large numbers of ferrous iron granules. The exceptions were the globus pallidus externa and interna, which contained light DMT1 staining, but large numbers of ferrous iron granules. The thalamus, subthalamic nucleus, and substantia nigra pars compacta contained neurons that were lightly stained for DMT1, but few or no iron granules. The high levels of DMT1 expression in some of the nuclei of the basal ganglia, particularly the caudate nucleus, putamen, and substantia nigra pars reticulata, may account for the high levels of iron in these regions.     

Comparative activities of tetanus and botulinum toxins

Using immunohistochemical methods we have studied the distribution of substance P fibers, terminals and perikarya throughout the basal ganglia of baboons and at selected levels of the human brain. Immunoreactivity in the substantia nigra pars reticulata, internal segment of the globus pallidus and ventral pallidum was dense and of a characteristic, “woolly-fiber” morphology. The caudate nucleus and putamen contained sharply circumscribed patches of dense immunoreactivity superimposed on a moderately stained background. The external division of the globus pallidus displayed very little immunoreactivity. Two morphological types of immunoreactive cell bodies were present in the caudate nucleus, putamen and nucleus accumbens, and were clustered within the dense patches. The distribution of immunoreactive perikarya within the striatum differed from that reported for rats, as immunoreactive neurons were distributed evenly throughout the rostrocaudal extent rather than being concentrated in the rostral portions.     

Single-cell RT-PCR, in situ hybridization histochemical, and immunohistochemical studies of substance P and enkephalin co-occurrence in striatal projection neurons in rats

Single-cell RT-PCR studies in 3-4-week-old rats have raised the possibility that as many as 20% of striatal projection neurons may be a unique type that contains both substance P (SP) and enkephalin (ENK). We used single-cell RT-PCR, retrograde labeling, in situ hybridization histochemistry, and immunolabeling to characterize the abundance of this cell type, its projection target(s), and any developmental changes in its frequency. We found by RT-PCR that 11% of neurons containing either SP or ENK contained both in 4-week-old rats, while in 4-month-old rats SP/ENK colocalization was only 3%. SP-only neurons tended to co-contain dynorphin and ENK-only neurons neurotensin, while SP/ENK neurons tended to contain dynorphin. Single-cell RT-PCR showed SP/ENK co-occurrence in 4-week-old rats to be no more common among striatal neurons retrogradely labeled from the substantia nigra than among those retrogradely labeled from globus pallidus. Double-label in situ hybridization showed SP/ENK perikarya to be scattered throughout striatum, making up 8% of neurons containing either SP or ENK at 4 weeks, but only 4% at 4 months. Immunolabeling showed that presumptive striatal terminals in globus pallidus externus, globus pallidus internus and substantia nigra pars reticulata that colocalized SP and ENK were scarce. Terminals colocalizing SP and ENK were, however, abundant in the substantia nigra pars compacta. Thus, SP-only and ENK-only neurons make up the vast majority of striatal projection neurons in rats, the frequency of SP/ENK colocalizing striatal neurons is low in adult rats (3-4%), and SP/ENK colocalizing neurons primarily project to SNc but do not appear to be confined to striosomes.     

Quantitative imaging of substance P in the human brain using a brain mapping analyzer

The distribution of substance P (SP)-like immunoreactive neurons in the brains of aged normal human was analyzed quantitatively. Consecutive coronal sections in which the striatum and the substantia nigra were exposed widely, were obtained from the right hemisphere and stained immunohistochemically for SP. Each stained section was divided into approximately three million microareas and the immunohistochemical fluorescence intensity in each area was measured using a human brain mapping analyzer, which is a microphotometry system for analysis of the distribution of neurochemicals in a large tissue slice. These distributions are displayed in color and monochromatic graphics. In the analyzed brain regions, conspicuously intense SP-like immunoreactivity was observed in the substantia nigra and the internal segment of the globus pallidus. Within the substantia nigra, the SP-like immunoreactive intensity in the pars compacta was 25%, higher than that in the pars reticulata, and the distribution of melanin-containing neurons corresponded well to the distribution of the SP-containing structures. SP-like immunoreactive intensity in the internal segment of the globus pallidus, which was lower than that in the substantia nigra, was approximately twice as high as that in the external segment of the globus pallidus. Very intense immunoreactivity was localized at the most medial area of the internal segment of the globus pallidus. The SP-like immunoreactive intensity in the caudate nucleus and putamen was moderate, and the distribution was heterogeneous and observed in patches.     

Evidence that histochemically distinct zones of the primate substantia nigra pars compacta are related to patterned distributions of nigrostriatal projection neurons and striatonigral fibers

Summary A marked histochemical compartmentalization is visible in the substantia nigra of the squirrel monkey in sections stained for acetylcholinesterase (AChE). In nigral regions containing tyrosine hydroxylase-positive neurons, there are AChE-poor and AChE-rich zones, and many of the AChE-poor zones have the form of narrow fingers extending ventrally into an AChE-rich matrix (Jimenez-Castellanos and Graybiel 1987b). The study reported here was carried out to determine whether this histochemical heterogeneity of the primate's substantia nigra is related to the known differentiation within its pars compacta of subdivisions projecting respectively to the caudate nucleus and to the putamen. Retrograde and anterograde labeling in the substantia nigra was elicited by tracer injections placed in the caudate nucleus or putamen and was plotted in relation to patterns of AChE staining and tyrosine hydroxylase immunostaining. Much of the labeling observed was organized according to borders visible with AChE histochemistry: labeled nigral neurons (and afferent fibers) tended to be clustered precisely within the AChE-poor ventrally-extending fingers or to be situated outside these zones. However, projection neurons in these ventrally-extending fingers were not exclusively related either to the caudate nucleus or to the putamen. After injections in the caudate nucleus, labeled neurons were predominantly in the AChE-poor fingers in some cases, but predominantly in AChE-rich nigral zones outside them in other cases. Labeling in and out of the ventrally-extending fingers, and along the edges of the fingers, also occurred following different tracer injections in the putamen. These findings confirm the independent clustering of nigrostriatal neurons projecting respectively to the caudate nucleus and to the putamen. The plan of nigrostriatal connections additionally appears concordant with the histochemical compartmentalization of the substantia nigra that can be detected with acetylthiocholinesterase histochemistry.     

Peptidase-containing neurons in rat striatum

The effects of surgical lesions on peptidase activity have been studied in the striatonigral system of the rat brain. Knife cuts separating the anterior part of the caudate putamen from the globus pallidus resulted in a decrease in the activity of angiotensin-converting enzyme and alanyl aminopeptidase in both the globus pallidus and substantia nigra. The activity of nigral prolyl endopeptidase and leucyl aminopeptidase was also decreased. An increase in dipeptidyl aminopeptidase and arginyl endopeptidase activity was observed in both the caudate putamen and globus pallidus. These results suggest that the striatal neurons containing angiotensin-converting enzyme or alanyl aminopeptidase project to both the globus pallidus and substantia nigra, and the neurons containing prolyl endopeptidase and/or leucyl aminopeptidase project to the substantia nigra. Dipeptidyl aminopeptidase and arginyl endopeptidase are probably associated with glial function.     

Association of dopamine D1 and D2 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

To ascertain the cellular associations of the D1 and D2 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 vitro receptor autoradiography with [3H]SCH23390 (D1 radioligand) or [3H]spiroperidol (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 D1 and D2 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 cat's 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, D2 binding in the two pallidal segments declines by approximately 50%, but remains unchanged in the substantia nigra. Binding of the D1 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 D2 receptor-binding by 95% in the pars compacta and 40% in the pars reticulata, but have no effect on the concentration of D1 or D2 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, D1 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 D2 receptors are associated with striatal cells and their axons in pallidal structures. Nearly all D2 receptors in the substantia nigra are associated with dopamine neurons (autoreceptors). Finally, the heterogeneous patterns of D1 and D2 receptors in the striatum are a consequence of intrinsic neuronal distributions.     

A survey of substance P, somatostatin, and neurotensin levels in aging in the rat and human central nervous system

Levels of the neuropeptides substance P, somatostatin, and neurotensin were measured by radioimmunoassay in regions of the rat and human central nervous system (CNS) in aging. Somatostatin levels were significantly lower only in the corpus striatum of older rats. Substance P levels and neurotensin levels were generally stable with aging as were levels of somatostatin in regions other than the corpus striatum. In post-mortem human CNS tissues, no significant negative correlations of levels of the three peptides were observed with time to refrigeration or time to freezer for the samples. In the human CNS, there were no significant age-related alterations in substance P levels in frontal cortex, thalamus, hypothalamus, caudate nucleus, globus pallidus, or substantia nigra. There was a significant age-related decrease in substance P levels in the human putamen. This age-related decrease was not present in tissues from victims of Huntington's disease nor was there any striking difference in substance P levels as a function of duration of the disease. There were no significant age-related changes in somatostatin levels in human frontal cortex, caudate nucleus, putamen, medial globus pallidus, or substantia nigra. Among these same regions, there was a significant age-related decrease in neurotensin levels only in the pars compacta and pars reticulata of the human nigra. These results implicate neuropeptides in aging processes in certain regions of the CNS. There are differences between rats and humans with respect to neuropeptides in the aging process in the CNS. Deterioration of some neuropeptide pathways in and to human basal ganglia may be involved in the suspected functional deterioration of parts of the extrapyramidal system in aging.     

Regional distribution of cholecystokinin binding sites in macaque basal ganglia determined by in vitro receptor autoradiography

Cholecystokinin binding sites were labeled with [3H]cholecystokinin-8, [125I]cholecystokinin-33, and [125I]cholecystokinin-8 in major structures of macaque basal ganglia by in vitro receptor autoradiography. Analysis of autoradiograms revealed areas of heavy cholecystokinin binding in the neostriatum and substantia nigra that were set off, often quite sharply, from the adjacent globus pallidus and subthalamic nucleus where labeling was, by contrast, very light. Heavy label characterized the ventromedial and posterior parts of the caudate nucleus and adjacent putamen, binding was of moderate intensity in central areas of these regions, while, the dorsolateral margin of the head of the caudate and precommissural putamen, the dorsolateral one-third of the body of the caudate, and all but the most medial and ventral portions of the posterior putamen lateral to the pallidum were sparsely labeled. The pattern of cholecystokinin binding within the neostriatum was mottled; patches of reduced label stood out from the background of more prominent binding. However, those patches were only imperfectly correlated with the striosomal organization of both the caudate nucleus and putamen as revealed by acetylcholinesterase staining. Cholecystokinin binding in the substantia nigra was also intricately patterned. Moderately dense, vertically orientated bands of label were found in the dorsal one-third to half of the pars reticulata, providing a marked contrast to the near background levels in the ventral pars reticulata and overlying pars compacta. The present study shows that heavy cholecystokinin binding is confined to particular areas within the primate basal ganglia; the pattern of label within the substantia nigra and neostriatum can be linked to intrinsic and afferent connections of these structures. The confinement of binding sites to the dorsal pars reticulata suggests an association with dendrites of pars compacta neurons which invade this region; this interpretation is consistent with recent evidence of depletion of nigral cholecystokinin binding sites in macaques following chemical lesion of dopaminergic cells of the par compacta. In the neostriatum the distribution of binding shows overlap with its topographically organized corticostriatal innervation; portions of heavily labeled striatum coincide with regions innervated by association cortex of the frontal and temporal lobes, whereas regions of diminished binding correspond to areas innervated mainly by sensory and motor cortex. These latter findings suggest that cholecystokinin may have a particularly strong influence on cognitive aspects of striatal function.     

Striatal axons to the globus pallidus, entopeduncular nucleus and substantia nigra come mainly from separate cell populations in cat

The extent to which individual striatal neurons send collaterals to the globus pallidus, entopeduncular nucleus and substantia nigra in the cat brain was determined by double-retrograde tracing with rhodamine fluorescent latex microspheres in combination with either horseradish peroxidase or the fluorescent nuclear dye Diamidino Yellow. In each case, two of the three target nuclei were injected, each with a different tracer, until all three possible combinations of two had been obtained several times. In all cases in which the tracer encroaches upon a striatal target, there are cells labeled in the striatum of a size and shape that is consistent with the observation that they mainly belong to the category of medium striatal cells. Since the striatal projections to the globus pallidus, entopeduncular nucleus and substantia nigra are each topographically organized, the zones of cell-labeling within the striatum vary depending upon the portion of the target nucleus involved by the deposit. Thus, in many cases the fields of striatal cells containing one label overlap only slightly with those in which cells containing the other label occur. In other cases, however, there is extensive overlap of the striatal zones containing cells marked with either tracer. In all cases, very few double-labeled cells can be found, even where hundreds of cells labeled with either tracer are freshly intermingled. Doubly labeled cells occur somewhat more frequently in those cases where the tracers are placed in the entopeduncular nucleus and substantia nigra than in those with the other two combinations, suggesting that striatal axons branch more often to the entopeduncular nucleus and substantia nigra than to the globus pallidus and nigra or globus pallidus and entopeduncular nucleus. These findings confirm, that, in the cat as in the primate, the striatal axons to the substantia nigra arise from cells that are largely separate from the striatopallidal population, and further show that the axons to the globus pallidus and entopeduncular nucleus also emanate mainly from different cells.     

Organization and efferent projections of nucleus tegmenti pedunculopontinus pars compacta with special reference to its cholinergic aspects

In an attempt to evaluate the cellular organization and efferent projections of the nucleus tegmenti pedunculopontinus pars compacta, several experiments were performed in the rat. From measurements of neurons in the nucleus tegmenti pedunculopontinus pars compacta in Nissl-stained sections, the nucleus was observed to contain many large neurons which made it possible to demarcate this nucleus from surrounding pontomesencephalic reticular formation. Two other neuronal populations, medium and small neurons, were also seen in the nucleus tegmenti pedunculopontinus pars compacta. Detailed measurements showed that 90% by volume of all neurons in the nucleus tegmenti pedunculopontinus pars compacta were large and medium-sized neurons. After injections of [ ³H]leucine into the nucleus tegmenti pedunculopontinus pars compacta, transported label was observed in dorsally and ventrally coursing ascending fibers. The dorsally coursing fibers entered the centrolateral nucleus and centre median-parafascicular complex of the thalamus. The ventrally coursing fibers produced accumulation of silver grains in the ventral tegmental area, substantia nigra pars compacta, subthalamic nucleus, zona incerta and lateral hypothalamus. Crossed fibers of the nucleus tegmenti pedunculopontinus pars compacta were observed sparsely at the levels of the thalamus and posterior commissure, and to a greater degree through the supraoptic commissure of Meynert. Much less anterograde labeling was seen in the equivalent terminal sites on the contralateral side of the brain. By electron microscopic autoradiography major terminal sites of axons of the nucleus tegmenti pedunculopontinus pars compacta were examined in rats injected with [ ³H]leucine in the nucleus tegmenti pedunculopontinus pars compacta and later injected with horseradish peroxidase in the striatum and pallidum. Statistical data showed preferential radiolabeling of terminals forming asymmetrical synaptic contact with dendrites in the centrolateral nucleus, centre median-parafascicular complex and subthalamic nucleus. Apparent terminations in the substantia nigra pars compacta proposed in earlier studies and shown in the present light microscopic autoradiograms were not supported by this ultrastructural analysis. Several radiolabeled terminals of the asymmetrical type contacting horseradish peroxidase labeled dendrites in the thalamus confirmed direct input from the nucleus tegmenti pedunculopontinus pars compacta to the thalamostriate projection neurons. [ ³H]choline injections into the thalamus and subthalamic nucleus produced retrograde perikaryal labeling of large neurons in the nucleus tegmenti pedunculopontinus pars compacta. These neurons were unlabeled after [ ³H]choline injections in the substantia nigra. Other findings suggested retrograde transport of [ ³H]choline through cholinergic terminals as well as cholinergic fibers of passage. These data suggested a selective uptake mechanism for cholinergic fibers of passage.The results emphasize the cholinergic nature of the nucleus tegmenti pedunculopontinus pars compacta innervation of the thalamus and subthalamic nucleus. Large neurons in the nucleus tegmenti pedunculopontinus pars compacta seem responsible for this cholinergic innervation and probably provide the axon terminals making asymmetrical synapses in the thalamus and subthalamic nucleus as described above. In addition, large neurons as well as medium and small ones in the nucleus tegmenti pedunculopontinus pars compacta whose transmitters and exact destinations remain unknown send a number of axons through the supraoptic commissure of Meynert to innervate the contralateral subthalamic nucleus.     

Topographical projections from the posterior thalamic regions to the striatum in the cat,with reference to possible tecto-thalamo-striatal connections

Summary Projections from the posterior thalamic regions to the striatum were studied in the cat by the anterograde tracing method after injecting wheat germ agglutinin-horseradish peroxidase conjugate (WGA-HRP) into the caudalmost regions of the lateroposterior thalamic nucleus (caudal LP), suprageniculate nucleus (Sg) and magnocellular division of the medial geniculate nucleus (MGm). The results were further confirmed by the retrograde tracing method after injecting WGA-HRP into the regions of the caudate nucleus (Cd) and putamen (Put) where afferent fibers from the caudal LP, Sg and MGm were distributed. Fibers from the MGm, Sg or caudal LP were distributed mainly in the medial, middle or lateral part of the caudal half of the putamen (caudal Put), respectively. Although there was a considerable overlap, thalamostriatal fibers from the caudal LP terminated more caudally than those from the MGm. On the other hand, thalamocaudate fibers from the MGm, Sg and lateral part of the caudal LP overlapped with each other in the ventrolateral part of the caudal half of the caudate nucleus (caudal Cd). Fibers from the medial part of the caudal LP were distributed in the ventral part of the caudal Cd. In the superior colliculus (SC) of the cats with WGA-HRP injections in the caudal LP, retrogradely labeled neuronal cell bodies were mainly seen ipsilaterally in the superficial SC layer, and simultaneously, anterogradely labeled axon terminals were observed in the striatum. On the other hand, when WGA-HRP was injected into the Sg or MGm, labeled SC neurons were mainly located in the intermediate and deep SC layers. Thus, ascending impulses from the superficial SC layer may possibly be conveyed ipsilaterally via the caudal LP to the ventral and ventrolateral parts of the caudal Cd and the lateral part of the caudal Put, whereas those from the intermediate and deep SC layers may be relayed via the Sg and/or MGm to the ventrolateral part of the caudal Cd and the middle and medial parts of the caudal Put.Abbreviations AC anterior commissure - Am amygdaloid nucleus - Cd caudate nucleus - Ce centromedial nucleus - CL centrolateral nucleus - Cl claustrum - CM-Pf centre médian-parafascicular complex - CP cerebral peduncle - d deep SC layer - EC external capsule - Ep entopeduncular nucleus - GP globus pallidus - i intermediate SC layer - IC internal capsule - Ip interpeduncular nucleus - LG lateral geniculate nucleus - LP lateroposterior nucleus - MD mediodorsal nucleus - MG medial geniculate nucleus - MGm magnocellular division of MG - MGp principal division of MG - NBIC nucleus of brachium of inferior colliculus - O oculomotor nucleus - OT optic tract - Pom medial division of posterior group of thalamus - Pt pretectum - Pul pulvinar nucleus - Put putamen - Pv paraventricular nucleus of thalamus - R reticular nucleus of thalamus - Rh rhomboid nucleus - RN red nucleus - s superficial SC layer - SC superior colliculus - Sg suprageniculate nucleus - SN substantia nigra - SNpc pars compacta of SN - SNpr pars reticulata of SN - V lateral ventricle - VA ventroanterior nucleus - VL ventrolateral nucleus - VM ventromedial nucleus - WGA-HRP wheat germ agglutinin-HRP conjugate