Distribution of somatostatin immunoreactivity in the forebrain of the squirrel monkey: basal ganglia and amygdala.

The distribution of somatostatin immunoreactivity in the basal ganglia and amygdala of the squirrel monkey (Saimiri sciureus) was studied with specific polyclonal antibodies directed against somatostatin-28 and somatostatin-28(1-12). Both antibodies gave similar results with regard to the distribution of somatostatin-immunoreactive neuronal profiles. A moderately dense and highly heterogeneous network of somatostatin-positive fibers was observed throughout the striatum. A dorsoventral gradient of increasing immunoreactivity was noted in the striatum and the caudate nucleus was found to strain generally less intensely than the putamen. The immunoreactive fibers within the striatum were mostly thin and varicose and formed patches corresponding to the striosomes, as visualized on adjacent sections immunostained for calbindin. Although some somatostatin cell bodies rimmed the striosomes, most of the positive cells were rather uniformly scattered in the striatum. These medium-sized cells were significantly smaller in the caudate nucleus (93 microns2, S.D. = 26 microns2) than in the putamen (122 microns2, S.D. = 39 microns2), but their density was significantly higher in the caudate nucleus (29.7 cells/mm2, S.D. = 8.8 cells/mm2) than in the putamen (20.5 cells/mm2, S.D. = 7.0 cells/mm2). The nucleus accumbens stained moderately and positive cell bodies were evenly dispersed throughout this structure. In contrast, the olfactory tubercle displayed a heavily stained neuropil but positive neurons were encountered only in its polymorph layer. In the sublenticular region, dense fiber plexuses appeared in register with nonreactive cell clusters of the nucleus basalis of Meynert and of the nucleus of the anterior commissure. More caudally, a dense bundle of positive fibers was observed at the level of the ansa lenticularis, the inferior thalamic peduncle, and the adjoining bed nucleus of the stria terminalis. Several fibers contributing to this bundle were of the woolly type. Woolly fibers also coursed in the substantia innominata between the ventral aspect of the globus pallidus and the optic tract, and ascended in the internal medullary lamina separating the internal and external segments of the globus pallidus. Somatostatin-immunoreactive cell bodies were uniformly scattered throughout the substantia innominata. The various nuclei of the amygdala showed a wide range of immunoreactivity. The central nucleus was lightly reactive, whereas the intercalated masses displayed a moderate staining. A dorsoventral gradient of immunostaining was noted in the ventrolateral portion of the amygdala, the lateral nucleus being moderately to densely stained and the basal nucleus very lightly to lightly immunoreactive.(ABSTRACT TRUNCATED AT 400 WORDS)     

Heterogeneous distribution of the limbic system-associated membrane protein in the caudate nucleus and substantia nigra of the cat

The limbic system-associated membrane protein is a glycoprotein selectively associated, in the adult, with dendrites and cell bodies of neurons of the limbic system and related brain regions. In the present study, the distribution of the limbic system-associated membrane protein was studied by immunohistochemistry in the caudate nucleus and substantia nigra of the cat to determine how its expression relates to the compartmentalization of these areas. In all areas of the caudate nucleus, the pattern of limbic system-associated membrane protein immunoreactivity was highly heterogeneous, labeling zones that were in register with areas expressing neurochemical markers that classically identify striosomes. The extrastriosomal matrix exhibited low levels of staining. The results show that the limbic system-associated membrane protein is expressed by neurons within the target areas (striosomes) of subsets of limbic afferents (originating mainly from the basolateral nucleus of the amygdala and the prefrontal cortex), whereas regions of the caudate nucleus (extrastriosomal matrix) receiving inputs from other subdivisions of the limbic system, such as the cingulate cortex and the ventral tegmental area, contain relatively low levels of limbic system-associated membrane protein immunoreactivity. Thus the expression of this antigen may reflect the targeting of specific groups of limbic afferents to regions that are intimately associated with distinct components of the limbic system.

The presence of limbic system-associated membrane protein in neurons of the substantia nigra pars compacta does not appear to be related to the presence or absence of the protein in their striatal target areas. In the substantia nigra, immunoreactivity to the limbic system-associated membrane protein was intense in the cell-sparse zone of the pars compacta, an area known to project to the extrastriosomal matrix of the caudate nucleus. This contrasted with the absence of immunostaining in areas containing dense clusters of dopaminergic neurons (densocellular zone), which project to the limbic system-associated protein-rich striosomes. By analogy to findings in the caudate nucleus and in other brain areas, the results suggest that subgroups of nigral dopaminergic neurons identified on the basis of their terminal fields in the caudate nucleus, may also differ in their limbic afferents.     

Co-expression of neuropeptides in the cat's striatum: an immunohistochemical study of substance P, dynorphin B and enkephalin.

The expression of tachykinin-like and opioid-like peptides was studied in medium-sized neurons of the caudate nucleus in tissue from adult cats pretreated with colchicine. Two methods, a serial thin-section peroxidase-antiperoxidase technique and a two-fluorochrome single-section technique, were applied. Quantitative estimates were made mainly with the peroxidase-antiperoxidase method. The numbers of neurons expressing substance P-like, dynorphin B-like, and enkephalin-like immunoreactivity were recorded in regions identified, respectively, as striosomes and extrastriosomal matrix. Striosomes were defined by the presence of clustered substance P-positive and dynorphin B-positive neurons and neuropil. Tests for the co-existence of enkephalin-like peptide and glutamate decarboxylase-like immunoreactivity were also made with the peroxidase-antiperoxidase method. Co-expression of substance P-like and dynorphin B-like immunoreactivities was the rule both in striosomes and in the matrix. In striosomes, substance P-like immunoreactivity was found in 96% of dynorphin B-immunoreactive neurons, and in the matrix 89% of dynorphin B-positive cells contained substance P-like immunoreactivity. Substance P/dynorphin B-positive neurons corresponded to over half (57%) of the neurons in striosomes but only 39% of the neurons in the matrix. Both in the matrix and in striosomes, about two-thirds of all neurons (63% and 65%, respectively) were identified as enkephalin-positive. Among all substance P/dynorphin B-positive medium-sized neurons, 76% also contained enkephalin-like antigen. The enkephalin-positive neurons characterized by triple peptide co-existence (enkephalin/substance P/dynorphin B) represented a mean of 63% of striosomal enkephalin-positive neurons (41% of all striosomal neurons) and 35% of matrical enkephalin-positive neurons (26% of all matrical neurons). Finally, nearly all enkephalin-positive neurons were immunoreactive for glutamate decarboxylase, and therefore probably GABAergic, but only about half the glutamate decarboxylase-positive population was enkephalin-immunoreactive. These findings suggest that neuropeptides from three distinct precursors may be co-localized in single medium-sized neurons in the striatum, and that the differential patterns of co-expression of substance P-like, dynorphin B-like, and enkephalin-like peptides may confer functional specializations upon subpopulations of GABAergic neurons giving rise to the efferent projections of the striatum. The linked expression of substance P-like and dynorphin B-like peptides in single neurons both in striosomes and matrix suggests that some regulatory mechanisms controlling peptide expression apply regardless of compartment.(ABSTRACT TRUNCATED AT 400 WORDS)     

Compartmental origins of striatal efferent projections in the cat

Injections of the retrograde tracer, wheat germ agglutinated-horseradish peroxidase were placed in the substantia nigra, in adjoining dopamine-containing cell groups A8 and A10, and in the internal and external parts of the pallidal complex of 20 cats in order to identify the compartmental origins of striatal efferent projections to the pallidum and midbrain. Patterns of retrograde cell-labeling in the caudate nucleus were analysed with respect to its striosomal architecture as detected in sections stained for acetylcholinesterase. Where possible, a similar compartmental analysis of cell-labeling in the putamen was also carried out. In 15 cats anterograde labeling in the striatum was studied in the sections stained with wheat germ agglutinated-horseradish peroxidase or in autoradiographically treated sections from cases in which [35S]methionine was mixed with the wheat germ agglutinated-horseradish peroxidase in the injection solution. Predominant labeling of projection neurons lying in striosomes (usually with some labeling of dorsomedial matrix neurons) occurred in a subset of the cases of nigral injection, including all cases (n = 9) in which the injection sites were centered in the densocellular zone of the substantia nigra pars compacta [Jiménez-Castellanos J. and Graybiel A. M. (1987) Neuroscience 23, 223-242.] Dense labeling of neurons in the extrastriosomal matrix, with at most sparse labeling of striosomal neurons, occurred in all cases of pallidal injection (n = 8) and in two cases of nigral injection in which the injection sites were lateral and anterior to the densocellular zone. Mixed labeling of striosomal and matrical neurons occurred in a third group of cases in which the injection sites were lateral to the densocellular zone but close to it. In a single case with an injection site situated in the pars lateralis of the substantia nigra, there was preferential labeling of striosomal neurons in the caudal caudate nucleus but widespread labeling of neurons in both striosomes and matrix in the putamen. A second type of compartmental ordering of projection neurons was found in the extrastriosomal matrix of the striatum. In cases of pallidal and nigral injection, there were gaps in cell labeling that did not match striosomes precisely, and often clusters of labeled cells appeared that did not correspond to acetylcholinesterase-poor striosomes but, instead, to patches of matrix. Especially prominent were clusters beside striosomes. There was a topographic ordering of striatal projection neurons both in the striosomes and in the extrastriosomal matrix according to their dorsoventral and latitudinal positions.(ABSTRACT TRUNCATED AT 400 WORDS)     

Quantitative electron microscopic study of immunoreactive somatostatin axons in the rat neostriatum

Various features of immunoreactive somatostatin axons including bouton size, synaptic length, the type of synapse formed (symmetric or asymmetric) and postsynaptic target, were examined at the ultrastructural level in the caudate nucleus. These features were compared to those of unlabeled axons in the surrounding caudate neuropil. Results showed that immunoreactive somatostatin axons make relatively short-surfaced, symmetric contacts, mostly with dendritic shafts whereas the majority of unlabeled axons form long-surfaced, asymmetric synapses with dendritic spines. Observations indicate that immunoreactive somatostatin axons belong to a sparse and homogeneous population of axons, have features corresponding to those of intrinsic caudate neurons, and synapse with caudate spiny cells. These findings are consistent with earlier speculation that immunoreactive somatostatin axons in caudate arise from a population of aspiny interneurons which have previously been identified to contain the peptide.     

Cellular substrate of the histochemically defined striosome/matrix system of the caudate nucleus: a combined Golgi and immunocytochemical study in cat and ferret

In order to learn what morphological substrate might underly the histochemical compartments of the neostriatum, sections of the caudate nucleus and the putamen of cats and ferrets were stained immunocytochemically with antisera directed against several neuropeptides and transmitter-related enzymes and were then Golgi-impregnated. Adjacent sections were stained to reveal acetylcholinesterase activity to identify the acetylcholinesterase-poor striosomes. The immunostaining produced by several of the antibody preparations was in register with the acetylcholinesterase-poor striosomes but the most prominent staining of these zones occurred with the antibodies directed against substance P. The striosomes were delineated by intense substance P-immunostaining of neuronal perikarya and dendrites, and in the rostral and dorsal caudate nucleus the boundary between substance P-immunostained and extrastriosomal matrix was abrupt. For these reasons we analysed Golgi-impregnated neurons in sections immunostained for substance P in order to assess the influence of the chemically defined striosomal architecture on the position and dendritic arborization of neurons located both within the striosomes and within the extrastriosomal matrix. The most commonly impregnated neurons were of the medium-size densely spiny class. Those that were present within the striosomes and lay within one dendritic radius of the boundary were divided into two types: (1) neurons whose dendritic arborization was apparently not influenced by the boundary and (2) neurons whose dendritic arborization was markedly influenced by the boundary. For neurons of the latter type, dendrites either emerged from the parts of the perikaryon away from the boundary, so avoiding crossing it, or they exhibited abrupt changes in their course, apparently to avoid crossing the boundary. Spiny neurons located in the extrastriosomal matrix but close to the striosomal boundary had dendrites that were either influenced by, or not influenced by the compartmental boundary. We conclude that there is a specific cytoarchitecture underlying the histochemical compartments of the neostriatum and that different sub-populations of medium-size spiny neurons underly (1) the segregation of information flow in striosomes and the extrastriosomal matrix and (2) communication between striosomes and the extrastriosomal matrix.     

Subdivisions of the dopamine-containing A8-A9-A10 complex identified by their differential mesostriatal innervation of striosomes and extrastriosomal matrix

The mesostriatal projections from the dopamine-containing cells groups A8, A9 and A10 have been studied in the cat in relation to the histochemical compartments known to exist in the striatum. In order to do this, we made stereotaxic injections in the substantia nigra of either [3H]proline-[3H]leucine, [35S]methionine, wheat germ agglutinin-horseradish peroxidase, or the two last tracers combined, and compared the location of anterograde labeling in the striatum to the locations of striosomes and extrastriosomal matrix identified by their low or high content, respectively, of the enzyme acetylcholinesterase. A discrete innervation of dorsolateral striosomes by a caudal densocellular subdivision of the substantia nigra pars compacta was found. This densocellular zone of the pars compacta was readily identifiable in sections stained for tyrosine hydroxylase-like immunoreactivity and corresponded to the uniquely acetylcholinesterase-poor zone detected in the substantia nigra pars compacta in serially adjacent sections stained for this enzyme. Selective anterograde labeling of the extrastriosomal matrix occurred in cases with injection sites centered in cell group A8. Tracer deposits in cell group A10 also elicited a preferential labeling of the extrastriosomal matrix, but this innervation was sparse compared to the prominent labeling of fibers in the ventral striatum. An almost exclusive innervation of caudal and ventral striosomes of the head of the caudate nucleus occurred after a deposit of tracer in the pars lateralis of the substantia nigra. Mixed labeling of striosomes and matrix occurred with injection sites centered in the rostral, cell-sparse part of the pars compacta of the substantia nigra. Clusters of tyrosine hydroxylase-immunoreactive neurons within this zone, most likely representing finger-like extensions of the caudal densocellular zone of the pars compacta, might have accounted for part of the striosomal labeling in these cases. We conclude that different subdivisions of the A8-A9-A10 dopamine-containing cell complex of the cat's mesencephalon project preferentially to striosomes or to extrastriosomal matrix. On this basis we suggest that there may be different functional channels in the mesostriatal projection, including, from cell group A8, a channel providing dopaminergic modulation of sensorimotor processing in the striatal matrix, and, from the densocellular zone of the substantia nigra pars compacta, a channel leading to limbic-related mechanisms represented in the striosomal system.     

Three-dimensional organization of the striosomal compartment and patchy distribution of striatonigral projections in the matrix of the cat caudate nucleus

Acetylcholinesterase staining on successive frontal or sagittal sections was used to determine the three-dimensional organization of the striosomal and matrix compartments in the adult cat caudate nucleus. Reconstruction drawings of the acetylcholinesterase-poor zones (striosomes) indicated that the striosomal compartment is a labyrinthine network organized in the rostrocaudal and mediolateral axis which is reproducible from one animal to another. Four main anteroposterior channels converging in the mediorostral pole of the caudate nucleus were distinguished. Seven to eight diagonally oriented channels crossing the previous ones were seen also in the mediolateral axis on the central core of the caudate nucleus. The pattern of organization of the numerous and tortuous striosomal channels was more complicated medially, while the lateral part of the caudate nucleus was represented mainly by the matrix compartment. In addition, a sub-compartmentation of the matrix was demonstrated by retrograde tracing studies made by injecting either horseradish peroxidase-wheat germ agglutinin, [14C]amino acids or a mixture of horseradish peroxidase-wheat germ agglutinin and [14C]amino acids in several areas of the substantia nigra pars reticulata. Labelled patches were seen with both tracers, their topographical localization depended on the nigral injection site but reconstruction analysis indicated that the populations of cells which innervate the substantia nigra pars reticulata originate in the two third lateral parts of the caudate nucleus all along its rostrocaudal extension. Examination of horseradish peroxidase-wheat germ agglutinin labelled cells indicated that not all cells were labelled in patches suggesting a further sub-compartmentation of these patches. Finally, a comparison of the topographical distributions of labelled patches and of striosomes revealed that most patches were located in the extrastriosomal matrix.     

Correspondence between the dopamine islands and striosomes of the mammalian striatum

During the development of the mammalian striatum, the early-forming dopamine innervation is broken up into macroscopic patches called "dopamine islands". These express high tyrosine hydroxylase-like immunoreactivity and are also rich in acetylcholinesterase activity. The mature striatum has prominent macroscopic compartments called "striosomes" that were first characterized by their low acetylcholinesterase activity and since have been related to heterogeneities in striatal input-output organizations. This report describes two sets of experiments designed to determine the relationship between the dopamine islands and the striosomes. The distributions of striatal tyrosine hydroxylase-like immunoreactivity and acetylcholinesterase activity were first compared in a series of kittens and young cats ranging in age from 1-228 postnatal days. During this time, the pattern of tyrosine hydroxylase-like immunoreactivity changed from islandic (patchy) to diffuse, and the pattern of acetylcholinesterase staining changed from one of acetylcholinesterase-rich patches to one of acetylcholinesterase-poor striosomes. The dopamine islands were in register with the acetylcholinesterase-poor patches at early developmental stages and at later stages the islands matched striosomes. These observations establish a correspondence between the dopamine islands and striosomes and demonstrate that the acetylcholinesterase-rich patches of the immature caudate nucleus become the acetylcholinesterase-poor striosomes of the adult. In a second set of experiments, cat fetuses were exposed to [3H]thymidine at embryonic days 22-29 in order to label the clustered subpopulations of striatal neurons known from previous experiments to lie in striosomes [Graybiel and Hickey (1982) Proc. natn. Acad. Sci. U.S.A. 79, 198-202]. The [3H]thymidine-labeled brains were examined at late fetal (embryonic days 50-52), early postnatal (days 1-21) and later postnatal (days 62-199) ages. The clusters of [3H]thymidine-labeled neurons were aligned with tyrosine hydroxylase-rich, acetylcholinesterase-rich patches early in development, and with acetylcholinesterase-poor striosomes at later stages. There were marked dorsoventral differences in the intensity of tyrosine hydroxylase-like immunoreactivity in the dopamine islands and this was confirmed in neonatal rats. A "dorsal islandic system" was defined as having crisp, highly immunoreactive islands; ventrally, regions of low and medium tyrosine hydroxylase-like immunoreactivity formed a mosaic.(ABSTRACT TRUNCATED AT 400 WORDS)     

Striosomes and extrastriosomal matrix contain different amounts of immunoreactive choline acetyltransferase in the human striatum

Cholinergic neuropil and cell bodies were identified in the human striatum by immunohistochemistry carried out with a polyclonal antibody raised against choline acetyltransferase (ChAT). The cholinergic neuropil was not uniformly distributed in the striatum, and especially in the caudate nucleus ChAT-poor zones corresponding to acetylcholinesterase (AChE)-poor striosomes were identified. Striosomal organization of ChAT-positive neuropil was also detected in striatal tissue from patients who had suffered parkinsonian and choreic disorders.     

Dopaminergic innervation of human basal ganglia

This paper summarises the results of some of our recent tyrosine hydroxylase (TH) immunohistochemical studies of the dopaminergic innervation of the human basal ganglia. It also reports new findings on the presence of TH-immunoreactive (ir) neurons in the striatum. Our data show the existence of nigrostriatal TH-ir axons that provide collaterals arborizing in the globus pallidus and subthalamic nucleus. These thin and varicose collaterals emerge from thick and smooth axons that course along the main output pathways of the basal ganglia, including the ansa lenticularis, the lenticular fasciculus and Wilson's pencils. We postulate that this extrastriatal innervation, which allows nigral dopaminergic neurons to directly affect the pallidum and subthalamic nucleus, plays a critical role in the functional organisation of human basal ganglia. The TH-ir fibres that reach the striatum arborize according to a highly heterogeneous pattern. At rostral striatal levels, numerous small TH-poor zones embedded in a TH-rich matrix correspond to calbindin-poor striosomes and calbindin-rich extrastriosomal matrix, respectively. At caudal striatal levels, in contrast, striosomes display a TH immunostaining that is more intense than that of the matrix. A significant number of small, oval, aspiny TH-ir neurons scattered throughout the rostrocaudal extent of the caudate nucleus and putamen, together with a few larger, multipolar, spiny TH-ir neurons lying principally within the ventral portion of the putamen, were disclosed in human. This potential source of intrinsic striatal dopamine might play an important role in the functional organisation of the human striatum, particularly in case of Parkinson's disease.     

Compartmental organization of the peptide network in the human caudate nucleus

The mammalian striatum may be divided into a striosomal compartment and a surrounding matrix region. We have examined the distribution of leucine enkephalin (LENK) and substance P (SP) immunoreactivity in relation to striosomes defined by calbindin-D (CABD) staining in alternate 70 μm serial sections from the human caudate nucleus. The distribution of LENK immunoreactivity showed a transition from dorsal to ventral striatum: dorsally, LENK-rich patches were present in a lightly stained matrix; mid-ventrally, annular patches of LENK staining with a lighter core were seen. These patches corresponded to striosomal regions defined by CABD-poor zones. In contrast, in the ventral caudate and nucleus accumbens, LENK-poor zones matched CABD-defined striosomes. CABD staining in the matrix was intense in the dorsal caudate, diminishing ventrally. SP-rich zones in dorsal caudate and SP-poor areas in the mid-ventral region overlapped striosomes. In the ventromedial sector, the SP staining pattern was complex and did not consistently correlate with striosomes. Computer-assisted three-dimensional reconstruction of the striosomal system in the human, based on regions of either high LENK or low CABD immunoreactivity, revealed the existence of considerable long-range order. Patches appeared aligned over several millimeters to form long, horizontal structures in the caudate nucleus, with occasional orthogonal interconnecting crossbridges. Our results are in accord with previous work in the human and in other species. These three-dimensional networks are strikingly similar across individuals and may relate to the segregation of and interactions between striatal circuits.     

Thalamic inputs to striatal interneurons in monkeys: synaptic organization and co-localization of calcium binding proteins.

Recent studies indicate that extrinsic inputs from sensorimotor regions of the cerebral cortex and the centromedian intralaminar thalamic nucleus terminate preferentially upon specific subpopulations of striatal output neurons in monkeys. The objective of the present study was to verify whether this specificity of innervation also characterizes the synaptic interactions between thalamic inputs from the centromedian nucleus and the four major populations of striatal interneurons. This was achieved by double labelling techniques at the electron microscope level, combining the anterograde transport of biotinylated-dextran amine with the immunostaining for specific markers of striatal interneurons (somatostatin, parvalbumin, choline acetyltransferase and calretinin). Injections of biotinylated-dextran amine in the centromedian nucleus led to dense bands of anterograde labelling which, in double immunostained sections, largely overlapped with the four populations of interneurons in the post-commissural region of the putamen. In the electron microscope, biotinylated-dextran amine-containing terminals formed asymmetric axo-dendritic synapses with somatostatin-, parvalbumin-, and choline acetyltransferase-containing elements. However, synapses between anterogradely labelled terminals and calretinin-positive neurons were not found. In sections processed to localize biotinylated-dextran amine and parvalbumin or calretinin, double-labelled terminals (biotinylated-dextran amine/parvalbumin and biotinylated-dextran amine/calretinin), morphologically similar to thalamostriatal boutons, were found in the striatum indicating that calcium binding proteins may be expressed by thalamostriatal neurons. To test this possibility, we combined the retrograde transport of lectin-conjugated horseradish peroxidase from the putamen with parvalbumin and calretinin immunostaining and found that, indeed, most of the retrogradely labelled cells in the centromedian nucleus displayed parvalbumin and calretinin immunoreactivity. Moreover, co-localization studies revealed that calretinin and parvalbumin co-exist in single neurons of the centromedian nucleus. In conclusion, striatal interneurons immunoreactive for somatostatin, parvalbumin and choline acetyltransferase, but not those containing calretinin, receive strong inputs from the centromedian nucleus in monkeys. Moreover, our findings indicate that parvalbumin and calretinin co-exist in individual thalamostriatal neurons. In combination with our previous data, these results suggest that thalamic information may be conveyed to striatal projection neurons both, directly via excitatory synaptic inputs, or indirectly via striatal interneurons. The relative importance of those direct and indirect thalamic influences upon the activity of striatal output neurons remains to be established.     

The striatal mosaic in primates: patterns of neuropeptide immunoreactivity differentiate the ventral striatum from the dorsal striatum.

Patterns of immunoreactivity for calcium-binding protein, tyrosine hydroxylase and four neuropeptides in the ventral striatum (nucleus accumbens, olfactory tubercle and ventromedial parts of the caudate nucleus and putamen) were compared to patterns of these markers in the dorsal striatum (the majority of the neostriatum) in rhesus monkey. The striatal mosaic was delineated by calcium-binding protein and tyrosine hydroxylase immunoreactivities. Both markers were found preferentially in the matrix of the dorsal striatum. The mosaic configurations of tyrosine hydroxylase, but not calcium-binding protein immunoreactivity, were similar in dorsal and ventral striatal regions. Substance P and leucine-enkephalin were not distributed homogeneously; distinct types and the prevalence of patches of substance P and leucine-enkephalin immunoreactivity distinguish the dorsal striatum from the ventral striatum and distinguish the caudate nucleus from the putamen. In the dorsal striatum, substance P and leucine-enkephalin patches consist of dense islands of immunoreactive neurons and puncta or clusters of immunoreactive neurons marginated by a dense rim of terminal-like puncta; the matrix was also enriched in leucine-enkephalin-immunoreactive neurons but contained less substance P-immunoreactive neurons. Patches were more prominent in the caudate nucleus than in the putamen. In the caudate, compartments low in tyrosine hydroxylase and calcium-binding protein immunoreactivities corresponded to cytologically identified cell islands and to patches enriched in substance P and leucine-enkephalin. These patches had a discrete infrastructure based on the location of substance P and leucine-enkephalin-immunoreactive neurons and terminals. In the ventral striatum, patches that showed low levels of substance P and leucine-enkephalin immunoreactivities were embedded in a matrix rich in immunoreactive cell bodies, fibers and terminals. In the accumbens, regions showing little tyrosine hydroxylase were in spatial register with patches low in substance P and leucine-enkephalin. Neurotensin- and somatostatin-immunoreactive neurons or processes were also compartmentally organized, particularly in the ventral striatum. Neurotensin-immunoreactive neurons were present predominantly in the nucleus accumbens but not in the dorsal striatum. Some regions enriched in neurotensin immunoreactivity were spatially registered with zones low in tyrosine hydroxylase, substance P and zones enriched in leucine-enkephalin. Areas enriched in somatostatin-immunoreactive processes overlapped with both tyrosine hydroxylase-rich and -poor regions in the ventral striatum. Our results show that the chemoarchitectonic topography of the striatal mosaic is different in the dorsal and ventral striatum of rhesus monkey and that the compartmental organization of some neurotransmitters/neuropeptides in the ventral striatum is variable and not as easily divisible into conventional patch and matrix regions as in the dorsal striatum.(ABSTRACT TRUNCATED AT 400 WORDS)     

Compartmental origins of the striatopallidal projection in the primate.

The organization of striatopallidal projection neurons in the primate was studied by injecting horseradish peroxidase conjugated with wheat germ agglutinin and fluorescent markers (latex microspheres, Fluorogold, Diamidino Yellow or Nuclear Yellow) into the globus pallidus of 20 adult squirrel monkeys (Saimiri sciureus). Single injections of horseradish peroxidase conjugated with wheat germ agglutinin were placed so as to involve predominantly either one or both pallidal segments. In the double-tracer experiments, fluorescent tracer injections were centered in the external pallidum and deposits of horseradish peroxidase conjugated with wheat germ agglutinin were made in the internal pallidum. In control cases, injections were made in nearby parts of the internal capsule or striatum. Distributions of retrogradely labeled neurons in the striatum were analysed in relation to its striosomal architecture as demonstrated by histochemistry and immunohistochemistry. Three principal findings emerged. (1) Both the external and the internal segments of the primate pallidum receive input from both the caudate nucleus and the putamen, but different sets of striatal cells within these nuclei project to the two segments. (2) The striatopallidal projection in the primate originates mainly in the extrastriosomal matrix, although striosomes in the fields of labeling almost always contain some labeled neurons. (3) Heterogeneous groupings of striatopallidal projection neurons exist in the matrix and appear to be parts of three-dimensional projection-neuron arrays. We conclude that in the primate, separate lines of conduction lead from the striatum to the external and the internal pallidal segments, and raise the possibility that the cells of origin of these pathways form a mosaic in the extrastriosomal matrix.     

GABAergic interneurons and neuropil of the intralaminar thalamus: an immunohistochemical study in the rat and the cat,with notes in the monkey

Summary Immunohistochemistry using antibodies to glutamic acid decarboxylase (GAD) was used to investigate the intralaminar nuclei of the thalamus in rat, cat and monkey. Antibodies to gamma aminobutyric acid (GABA) were also used in the cat. Intralaminar immunoreactive cell bodies were not detected in the rat, but were clearly present in cat and monkey. In the latter species, GABA- or GAD-immunopositive perikarya were distributed throughout the anterior intralaminar nuclei, whereas in the posterior intralaminar complex they prevailed in the lateral part of the centre median nucleus and around the fasciculus retrofiexus. Measurements of the area of immunostained intralaminar cell bodies in cat and monkey indicated that they are represented by small neurons. Experiments in the cat, based on retrograde tracers injections involving large sectors of the frontal and parietal cortices and the head of the caudate nucleus, revealed that the GABA- or GAD-immunoreactive cells and the retrogradely labeled projection neurons represented two separate intralaminar cell populations, although the latter also included small cells. Considerable differences were observed in the immunoreactive GABAergic neuropil of the anterior and posterior intralaminar nuclei. Clusters of densely packed bouton-like immunoreactive elements were detected in the former structures in the rat, cat and monkey, and were especially evident in the central lateral nucleus; immunopositive varicose fibers and puncta were diffusely distributed in the posterior intralaminar structures. Taken together with data from the literature, the present findings indicate that in cat and monkey local circuit inhibitory cells regulate not only the activity of principal thalamic nuclei which project densely upon restricted cortical fields, but also of the intralaminar structures which are widely connected with the cerebral cortex and the striatum. Regional variations in the distribution of GABAergic fibers and terminals suggest major differences in the organization of inhibitory circuits and synaptic arrangements of the anterior and posterior intralaminar thalamus.Abbreviations CeM central medial nucleus - CL central lateral nucleus - CM-Pf centre median-parafascicular complex - fr fasciculus retrofiexus - Hb habenular complex - LD laterodorsal nucleus - LP lateroposterior nucleus - MD mediodorsal nucleus - Pc paracentral nucleus - PO thalamic posterior complex - PvA anterior paraventricular nucleus - Sm stria medullaris - VL ventral lateral nucleus - VP ventroposterior complex     

Mosaic distribution of phosphate-activated glutaminase-like immunoreactivity in the rat striatum.

The dorsal and ventral striatum of mammals has been known to be organized in a mosaic manner, referred to as "patches" and "matrix" of the caudatoputamen. The present study was primarily attempted in order to reveal the relationship of glutamatergic neuronal components to the mosaic organization in the rat striatum by using a monoclonal antibody to phosphate-activated glutaminase, a major synthetic enzyme of transmitter glutamate. Antibodies against glutamate decarboxylase and choline acetyltransferase were also used as the markers for GABAergic and cholinergic neuronal components, respectively. Glutaminase immunoreactivity was seen in a number of large- and a few medium-sized neurons in the caudatoputamen, nucleus accumbens and olfactory tubercle. The large neurons with glutaminase immunoreactivity were observed in the neuropil of the caudatoputamen and nucleus accumbens; glutaminase immunoreactivity was particularly marked in the neuropil of island-like patchy areas although it was seen throughout the neuropil of the nuclei. In the caudatoputamen, island-like areas with marked glutaminase immunoreactivity exhibited less marked choline acetyltransferase immunoreactivity than the surrounding background region, and were thus considered to correspond to the patches. The mosaic distribution of glutamate decarboxylase immunoreactivity in the caudatoputamen seemed identical with that of glutaminase immunoreactivity. However, in the nucleus accumbens, the mosaic pattern of neuropil labeling for glutaminase was neither consistent with that for glutamate decarboxylase nor that for choline acetyltransferase, suggesting the presence of non-GABAergic glutaminase-containing nerve terminals in the nucleus. In an attempt to clarify the origin of neuropil labeling for glutaminase in the striatum, lesions were made in the regions sending projection fibers to the caudatoputamen and nucleus accumbens. After placing lesions in the cerebral cortex, glutaminase immunoreactivity was decreased in neuropil of the caudatoputamen, but the mosaic pattern remained. Lesions which were placed in the intralaminar thalamic nuclei, amygdaloid body, globus pallidus or substantia nigra produced no substantial change in glutaminase immunoreactivity in the caudatoputamen and nucleus accumbens. After injection of kainic acid into the caudatoputamen or nucleus accumbens, glutaminase immunoreactivity in the neuropil of the affected regions was decreased to lose the mosaic pattern, indicating that neuronal components with glutaminase immunoreactivity in the neuropil of the patches were mainly of intrinsic origin. In summary, possible axon terminals containing glutaminase were observed with mosaic patterns in the caudatoputamen and nucleus accumbens, in which large cholinergic and medium-sized non-cholinergic neurons were immunoreactive for glutaminase. In the caudatoputamen, glutaminase immunoreactivity in neuropil was more marked in the patches than in the matrix.(ABSTRACT TRUNCATED AT 400 WORDS)     

Immunohistochemical evidence for coexistence of methionine-enkephalin and tyrosine hydroxylase in neurons of the locus coeruleus complex projecting to the spinal cord of the cat.

Previous studies have revealed the presence of pontospinal neurons with either methionine-enkephalin- or tyrosine hydroxylase-like immunoreactivity in the dorsolateral pontine tegmentum of the cat. Using a combined fast blue retrograde transport technique and simultaneous immunofluorescence histochemistry, the present study was designed to reveal the coexistence of enkephalin and tyrosine hydroxylase in cat coerulospinal neurons and to determine if and to what extent the coerulospinal pathway is heterogeneous. Fast blue-labelled neurons with tyrosine hydroxylase- and enkephalin-like immunoreactivities were found in the nucleus locus coeruleus, nucleus subcoeruleus, K?lliker-Fuse nucleus, and the medial and lateral parabrachial nuclei. Approximately 87% of tyrosine hydroxylase-like immunoreactive neurons had enkephalin-like immunoreactivity, whereas about 76% of the enkephalin-like immunoreactive neurons had tyrosine hydroxylase-like immunoreactivity. About 71% of all coerulospinal neurons exhibited both tyrosine hydroxylase- and enkephalin-like immunoreactivities. These findings indicate that coerulospinal activity may lead to spinal cord effects reflecting both norepinephrine and enkephalin activity in most cases but do not rule out each transmitter's isolated functions.     

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.