Projections to the putamen from neurons located in the white matter and the claustrum in the macaque

Continuing investigations of corticostriatal connections in rodents emphasize an intricate architecture where striatal projections originate from different combinations of cortical layers, include an inhibitory component, and form terminal arborizations which are cell-type dependent, extensive, or compact. Here, we report that in macaque monkeys, deep and superficial cortical white matter neurons (WMNs), peri-claustral WMNs, and the claustrum proper project to the putamen. WMNs retrogradely labeled by injections in the putamen (four injections in three macaques) were widely distributed, up to 10 mm antero-posterior from the injection site, mainly dorsal to the putamen in the external capsule, and below the premotor cortex. Striatally projecting labeled WMNs (WMNsST) were heterogeneous in size and shape, including a small GABAergic component. We compared the number of WMNsST with labeled claustral and cortical neurons and also estimated their proportion in relation to total WMNs. Since some WMNsST were located adjoining the claustrum, we wanted to compare results for density and distribution of striatally projecting claustral neurons (ClaST). ClaST neurons were morphologically heterogeneous and mainly located in the dorsal and anterior claustrum, in regions known to project to frontal, motor, and cingulate cortical areas. The ratio of ClaST to WMNsST was about 4:1 averaged across the four injections. These results provide new specifics on the connectional networks of WMNs in nonhuman primates, and delineate additional loops in the corticostriatal architecture, consisting of interconnections across cortex, claustralstriatal and striatally projecting WMNs.     

Laminar origins of visual corticocortical connections in the cat

The interconnections among visual areas in cat cortex were studied with respect to the specific laminae in which the cortically projecting neurons are located. Single injections of HRP were made through recording micropipettes into nine different visual areas. In 15 cortical areas the laminar distribution of neurons which were retrogradely filled with HRP was plotted. In this way we determined the laminar origins of the cortical projections to the nine separate cortical visual areas which were injected. There are three major observations. First, areas 17 and 18 are the only two visual areas in which layers II and III are the primary site of cortically projecting cells; in the other 13 areas the deeper layers of cortex provide a large percentage of such neurons. Second, within any one cortical area, cortically projecting neurons may be distributed among different layers; the specific layer depends upon the cortical target of those neurons. Third, any one cortical area receives projections from several different cortical layers, the specific layers being dependent upon the area from which the projection originates. An individual cortical area, therefore, contributes to several different cortical visual circuits, with each of these circuits defined by the laminar connections of its neurons.     

Pattern in the laminar origin of corticocortical connections

The laminar origin of cortical projections to the frontal cortex was studied in 17 adult rhesus monkeys with the use of the retrograde transport of horseradish peroxidase (HRP). The frontal regions injected with HRP extended from the posterior periarcuate region to the frontal pole. The architectonic boundaries of areas containing HRP-labeled neurons were determined from matched sections stained for the visualization of cell bodies, myelin, or acetylcholinesterase. The results showed that the laminar origin of both nearby and distant corticocortical projections was correlated with the architectonic differentiation of the regions giving rise to the projecting afferent fibers. Frontally directed projections from limbic cortices, which show a rudimentary laminar organization, emanated mainly from deep layers. On the other hand, projections from increasingly more differentiated cortices arose progressively from the upper (or supragranular) layers. This pattern was observed for projections originating along the axis of architectonic differentiation of the visual, somatosensory, auditory, motor, and prefrontal cortical systems. Thus, as the cortical architecture within each system changes from limbic areas toward the primary cortices, the origin of frontally directed projections shifts from predominantly infragranular to predominantly supragranular layers.     

Organization of thalamostriatal terminals from the ventral motor nuclei in the macaque

This study examines the organization of thalamostriatal projections from ventral tier nuclei that relay basal ganglia output to the frontal cortex. Although previous thalamostriatal studies emphasize projections from the intralaminar nuclei, studies in primates show a substantial projection from the ventral anterior (VA) and ventral lateral (VL) nuclei. These nuclei make up the main efferent projection from the basal ganglia to frontal cortical areas, including primary motor, supplementary, premotor, and cingulate motor areas. Functionally related motor areas of the frontal cortex and VA/VL have convergent projections to specific regions of the dorsal striatum. The distribution of VA/VL terminals within the striatum is crucial to understanding their relationship to motor cortical afferents. We placed anterograde tracer injections into discrete VA/VL thalamic areas. VA/VL thalamostriatal projections terminate in broad, rostrocaudal regions of the dorsal striatum, corresponding to regions innervated by functionally related cortical motor areas. The pars oralis division of VL projects primarily to the dorsolateral, postcommissural putamen, whereas the parvicellular VA targets more medial and rostral putamen regions, and the magnocellular division of VA targets the dorsal head of the caudate nucleus. Whereas these results demonstrate a general functional topography, specific VA/VL projections overlap extensively, suggesting that functionally distinct VA/VL projections may also converge in dorsal striatal areas. Within striatal territories, VA/VL projections terminate in a patchy, nonhomogeneous manner, indicating another level of complexity. Moreover, terminal fields contain both terminal clusters and scattered, long, unbranched fibers with many varicosities. These fiber morphologies resemble those from the cortex and raise the possibility that VA/VL thalamostriatal projections neurons have divergent connectional features.     

Relationship between the corticostriatal terminals from areas 9 and 46, and those from area 8A, dorsal and rostral premotor cortex and area 24c: an anatomical substrate for cognition to action

Our previous data indicate that there are specific features of the corticostriatal pathways from the prefrontal cortex. First, corticostriatal pathways are composed of focal, circumscribed projections and of diffuse, widespread projections. Second, there is some convergence between terminal fields from different functional regions of the prefrontal cortex. Third, anterior cingulate projections from area 24b occupy a large region of the rostral striatum. The goal of this study was to determine whether these features are also common to the corticostriatal projections from area 8A (including the frontal eye field; FEF), the supplementary eye field (SEF), dorsal and rostral premotor cortex (PMdr) and area 24c. Using a new approach of three-dimensional reconstruction of the corticostriatal pathways, along with dual cortical tracer injections, we mapped the corticostriatal terminal fields from areas 9 and 46, 8A-FEF, SEF, PMdr and 24b and c. In addition, we placed injections of retrogradely transported tracers into key striatal regions. The results demonstrated that: (i) a diffuse projection system is a common feature of the corticostriatal projections from different frontal regions; (ii) key striatal regions receive convergent projections from areas 9 and 46 and from areas 8A-FEF, SEF, PMdr and 24c, suggesting a potential pivotal role of these striatal regions in integrating cortical information; (iii) projections from area 24c, like those from area 24b, terminate widely throughout the striatum, interfacing with terminals from several frontal areas. These features of the corticostriatal frontal pathways suggest a potential integrative striatal network for learning.     

Differential laminar distribution of corticostriatal neurons in the prefrontal and pericruciate gyri of the dog

This investigation was undertaken to determine whether the laminar distribution of corticostriatal neurons differs between different anatomical and functional areas of the frontal lobe. Injections of lectin-conjugated horseradish peroxidase were made into the lateral, intermediate, and medial parts of the head of the caudate nucleus. After injections into the medial part of the caudate, most of the labeled neurons were found in the proreal and subproreal gyri of the prefrontal region. In both gyri, the majority of labeled cells were localized within layer V, and fewer neurons were located in the deeper part of layer III and in layers IV (proreal gyrus) and VI. No labeled neurons were noted in layer II. This distribution contrasted with that seen following injections into the intermediate and lateral parts of the caudate nucleus. In those cases, the majority of labeled cells were located in the pericruciate region (areas 4 and 6) and on the medial and lateral banks of the presylvian sulcus (paraorbital and internal composite areas, respectively). In the paraorbital and internal composite areas, the majority of labeled cells were still localized within layer V, but a larger percentage of labeled neurons were also noted in layer III. In areas 4 and 6, the laminar distribution shifted so that the majority of labeled cells were now located in layer III, with a widespread distribution of neurons throughout layers II-VI. These results indicate that corticostriatal neurons in the frontal lobe have different laminar distributions, depending on the particular anatomical or functional area in which they are located. The majority of neurons in the prefrontal region are located in infragranular layers, while those in the pericruciate region are located in supragranular layers.     

Characterization of transient cortical projections from auditory, somatosensory, and motor cortices to visual areas 17, 18, and 19 in the kitten

We have examined the anatomical features of ipsilateral transient cortical projections to areas 17, 18, and 19 in the kitten with the use of axonal tracers Fast Blue and WGA-HRP. Injections of tracers in any of the three primary visual areas led to retrograde labeling in frontal, parietal, and temporal cortices. Retrogradely labeled cells were not randomly distributed, but instead occurred preferentially at certain loci. The pattern of retrograde labeling was not influenced by the area injected. The main locus of transiently projecting neurons was an isolated region in the ectosylvian gyrus, probably corresponding to auditory area A1. Other groups of transiently projecting neurons had more variable locations in the frontoparietal cortex. The laminar distribution of neurons sending a transient projection to the visual cortex is characteristic and different from that of parent neurons of other cortical pathways at the same age. In the frontoparietal cortex, transiently projecting neurons were located mainly in layer 1 and the upper part of layers 2 and 3. In the ectosylvian gyrus, nearly all the neurons are located in layers 2 and 3. In addition, a few transiently projecting neurons are found in layer 6 and in the white matter. Transiently projecting neurons have a pyramidal morphology except for the occasional spindle-shaped cell of layer 1 and multipolar cells observed in the white matter. Anterograde studies were used to investigate the location of transient fibers in the visual cortex. Injections of WGA-HRP at the site of origin of transient projections gave rise to few retrogradely labeled cells in areas 17, 18, and 19, demonstrating that transient projections to these areas are not reciprocal. Although labeled axons were found over a wide area of the posterior cortex, they were more numerous over certain regions, including areas 17, 18, and 19, and absent from other more lateral cortical regions. Transient projecting fibers were present in all cortical layers at birth. Plotting the location of transient fibers in numerous sections and at all ages showed that these fibers are not more plentiful in the white matter than they are in the gray matter. We found no evidence that the white/gray matter border constituted a physical barrier to the growth of transient axons. Comparison of the organization of this transient pathway to that of other transient connections is discussed with respect to the development of the cortex.     

Corticostriatal and corticosubthalamic input zones from the presupplementary motor area in the macaque monkey: comparison with the input zones from the supplementary motor area

The presupplementary motor area (pre-SMA) is a cortical motor-related area which lies in the medial wall of the frontal lobe, immediately anterior to the supplementary motor area (SMA). This area has been considered to participate in the control of complex forelimb movements in a way different from the SMA. In an attempt to analyze the patterns of projections from the pre-SMA to the basal ganglia, we examined the distributions of pre-SMA inputs in the striatum and the subthalamic nucleus and compared them with the SMA input distributions. To detect morphologically the terminal fields from the pre-SMA and the forelimb region of the SMA, anterograde tracers were injected into such areas that had been identified electrophysiologically in the macaque monkey. Corticostriatal inputs from the pre-SMA were distributed mainly in the striatal cell bridges connecting the rostral aspects of the caudate nucleus and the putamen, as well as in their neighboring striatal portions. These input zones were located, with no substantial overlap, rostral to corticostriatal input zones from the SMA forelimb region. Corticosubthalamic input zones from the pre-SMA were almost localized in the medial aspect of the nucleus, where corticosubthalamic inputs from the SMA forelimb region were also distributed predominantly. However, the major terminal fields from the pre-SMA were centered ventrally to those from the SMA. The present results indicate that the corticostriatal and corticosubthalamic input zones from the pre-SMA appear to be segregated from the SMA-derived input zones. This implies the possibility of parallel processing of motor information from the pre-SMA and SMA in the cortico-basal ganglia circuit.     

Neurochemical,morphologic, and laminar characterization of cortical projection neurons in the cingulate motor areas of the macaque monkey

The primate cingulate gyrus contains multiple cortical areas that can be distinguished by several neurochemical features, including the distribution of neurofilament protein-enriched pyramidal neurons. In addition, connectivity and functional properties indicate that there are multiple motor areas in the cortex lining the cingulate sulcus. These motor areas were targeted for analysis of potential interactions among regional specialization, connectivity, and cellular characteristics such as neurochemical profile and morphology. Specifically, intracortical injections of retrogradely transported dyes and intracellular injection were combined with immunocytochemistry to investigate neurons projecting from the cingulate motor areas to the putative forelimb region of the primary motor cortex, area M1. Two separate groups of neurons projecting to area M1 emanated from the cingulate sulcus, one anterior and one posterior, both of which furnished commissural and ipsilateral connections with area M1. The primary difference between the two populations was laminar origin, with the anterior projection originating largely in deep layers, and the posterior projection taking origin equally in superficial and deep layers. With regard to cellular morphology, the anterior projection exhibited more morphologic diversity than the posterior projection. Commissural projections from both anterior and posterior fields originated largely in layer VI. Neurofilament protein distribution was a reliable tool for localizing the two projections and for discriminating between them. Comparable proportions of the two sets of projection neurons contained neurofilament protein, although the density and distribution of the total population of neurofilament protein-enriched neurons was very different in the two subareas of origin. Within a projection, the participating neurons exhibited a high degree of morphologic heterogeneity, and no correlation was observed between somatodendritic morphology and neurofilament protein content. Thus, although the neurons that provide the anterior and posterior cingulate motor projections to area M1 differ morphologically and in laminar origin, their neurochemical profiles are similar with respect to neurofilament protein. This suggests that neurochemical phenotype may be a more important unifying feature for corticocortical projections than morphology. © 1996 Wiley-Liss, Inc.     

Single-axon tracing study of corticostriatal projections arising from primary motor cortex in primates

The axonal projections arising from the forelimb area of the primary motor cortex (M1) in cynomolgus monkeys (Macaca fascicularis) were studied following microiontophoretic injections of biotinylated dextran amine under electrophysiological guidance. The microinjections were centered on layer V, and 42 anterogradely labeled corticofugal axons were reconstructed from serial frontal or sagittal sections with a camera lucida. Our investigation shows that the primate striatum receives both direct and indirect projections from M1. The direct corticostriatal projection is formed by axons that remain uniformly thin and unbranched throughout their sinuous trajectory to the ipsilateral striatum. They divide as they enter the dorsolateral sector of the post-commissural putamen, the so-called sensorimotor striatal territory. The indirect corticostriatal projection derives from a thin collateral emitted within the corona radiata by thick, long-range fibers that descend toward the brainstem. The collateral enters the putamen dorsomedially and remains unbranched until it reaches the dorsolateral sector of the putamen, where it breaks out into two to four axonal branches displaying small and equally spaced varicosities. Both direct and indirect corticostriatal axons branch moderately but occupy vast rostrocaudal striatal territories, where they appear to contact en passant several widely distributed striatal neurons. These findings reveal that, in contrast to current beliefs, the primate motor corticostriatal system is not exclusively formed by axons dedicated solely to the striatum. It also comprises collaterals from long-range corticofugal axons, which can thus provide to the striatum a copy of the neural information that is being conveyed to the brainstem and/or spinal cord.     

Distributed but convergent ordering of corticostriatal projections: analysis of the frontal eye field and the supplementary eye field in the macaque monkey.

The degree of parallel processing in frontal cortex-basal ganglia circuits is a central and debated issue in research on the basal ganglia. To approach this issue directly, we analyzed and compared the corticostriatal projections of two principal oculomotor areas of the frontal lobes, the frontal eye field (FEF) and the supplementary eye field (SEF). We first identified cortical regions within or adjacent to each eye field by microstimulation in macaque monkeys and then injected each site with either 35S-methionine or WGA-HRP conjugate. We analyzed the corticostriatal projections and also the interconnections of the pairs of cortical areas. We observed major convergence of the projections of the FEF and the SEF within the striatum, principally in the caudate nucleus. In cross sections through the striatum, both projections were broken into a series of discontinuous input zones that seemed to be part of complex three-dimensional labyrinths. Where the FEF and SEF projection fields were both present, they overlapped patch for patch. Thus, both inputs were dispersed within the striatum but converged with one another. Striatal afferents from cortex adjacent to the FEF and the SEF did not show convergence with SEF and FEF inputs, but did, in part, converge with one another. For all pairs of cortical areas tested, the degree of overlap in the corticostriatal projections appeared to be directly correlated with the degree of cortical interconnectivity of the areas injected. All of the corticostriatal fiber projections observed primarily avoided immunohistochemically identified striosomes. We conclude that there is convergence of oculomotor information from two distinct regions of the frontal cortex to the striatal matrix, which is known to project into pallidonigral circuits including the striatonigrocollicular pathway of the saccadic eye movement system. Furthermore, functionally distinct premotor areas near the oculomotor fields often systematically projected to striatal zones adjacent to oculomotor field projections, suggesting an anatomical basis for potential interaction of these inputs within the striatum. We propose that parallel processing is not the exclusive principle of organization of forebrain circuits associated with the basal ganglia. Rather, patterns of both convergence and divergence are present and are likely to depend on multiple functional and developmental constraints.     

Synaptic convergence of motor and somatosensory cortical afferents onto GABAergic interneurons in the rat striatum

Cortical afferents to the basal ganglia, and in particular the corticostriatal projections, are critical in the expression of basal ganglia function in health and disease. The corticostriatal projections are topographically organized but also partially overlap and interdigitate. To determine whether projections from distinct cortical areas converge at the level of single interneurons in the striatum, double anterograde labeling from the primary motor (M1) and primary somatosensory (S1) cortices in the rat, was combined with immunolabeling for parvalbumin (PV), to identify one population of striatal GABAergic interneurons. Cortical afferents from M1 and S1 gave rise to distinct, but partially overlapping, arbors of varicose axons in the striatum. PV-positive neurons were often apposed by cortical terminals and, in many instances, apposed by terminals from both cortical areas. Frequently, individual cortical axons formed multiple varicosities apposed to the same PV-positive neuron. Electron microscopy confirmed that the cortical terminals formed asymmetric synapses with the dendrites and perikarya of PV-positive neurons as well as unlabelled dendritic spines. Correlated light and electron microscopy revealed that individual PV-positive neurons received synaptic input from axon terminals derived from both motor and somatosensory cortices. These results demonstrate that, within areas of overlap of functionally distinct projections, there is synaptic convergence at the single cell level. Sensorimotor integration in the basal ganglia is thus likely to be mediated, at least in part, by striatal GABAergic interneurons. Furthermore, our findings suggest that the pattern of innervation of GABAergic interneurons by cortical afferents is different from the cortical innervation of spiny projection neurons.     

Thalamic projections to areas 3a, 3b,and 4 in the sensorimotor cortex of the mature and infant macaque monkey

Area 3a in the macaque monkey, located in the fundus of the central sulcus, separates motor and somatosensory cortical areas 4 and 3b. The known connections of areas 4 and 3b differ substantially, as does the information which they receive, process, and transfer to other parts of the central nervous system. In this analysis the thalamic projections to each of these three cortical fields were examined and compared by using retrogradely transported fluorescent dyes (Fast Blue, Diamidino Yellow, Rhodamine and Green latex microspheres) as neuron labels. Coincident labeling of projections to 2–3 cortical sites in each monkey allowed the direct comparison of the soma distributions within the thalamic space of the different neuron populations projecting to areas 3a, 3b, and 4, as well as to boundary zones between these cortical fields. The soma distribution ofthalamic neurons projecting to a small circumscribed zone (diameter = 0.5–1.0 mm) strictly within cortical area 3a (in region of hand representation) filled out a “territory” traversing the dorsal half of the cytoarchitectonically defined thalamic nucleus, VPLc (abbreviations as in Olszewski [1952] The Thalamus of the Macaca mulatta. Basel: Karger). This elongate, rather cylindrical, territory extended caudally into the anterior pulvinar nucleus, but not forward into VPLo. The rostrocaudal extent of the thalamic territory defining the soma distribution of neurons projecting to small zones of cortical area 3b was similar, but typically extended into the ventral part of VPLc, filling out a medially concavo-convex laminar space. Two such territories projecting to adjacent zones of areas 3a and 3b, respectively, overlapped and shared thalamic space, but not thalamic neurons. Contrasting with the 3a and 3b thalamic territories, the soma distribution of thalamic neurons projecting to a circumscribed zone in the nearby motor cortex (area 4) did not penetrate into VPLc, but instead filled out a mediolaterally flattened territory extending from rostral VLo, VLm, VPLo to caudal and dorsal VLc, LP, and Pulo. These territories skirted around VPLc. All three cortical areas (4, 3a, and 3b) also received input from distinctive clusters of cells in the intralaminar Cn.Md. It is inferred that, in combination, the thalamic territories in areas 3a, 3b, and 4 (and also area 1 and 2), which would be coactive during the execution of a manual task, constituted a lamellar space extending from VLo rostrally to Pul.o caudally. How Pul.o neuron populations relate to the more rostral populations within the same thalamic territory projecting to a localized cortical zone remains uncertain. Within the medially located territories the distribution of the neuron population in Pul.o was spatially continuous with the more rostral thalamic cells projecting to the same localized cortex, but in lateral thalamic territories these 2 populations were usually spatially discontinous. In the newborn macaque an orderly change in the territorial projections to localized zones in area 4, 3a, and 3b was also demonstable. However, thalamic nuclear projections were more expansive than in the mature animal. As well as the VPLc input, a third of the thalamic input to area 3a was now from VLo, VPLo, and VLm. Area 4 also had a significant input from VPLc, an input not observed in the mature macaque. © 1993 Wiley-Liss, Inc.     

Morphology and synaptic connections of crossed corticostriatal neurons in the rat

The neurons of origin of the bilateral corticostriatal projection arising from the medial agranular cortical field in rats were identified by antidromic activation from contralateral neostriatal stimulation. The same cells were tested for antidromic activation from the contralateral neocortex and for orthodromic responses to stimulation of neocortex of the contralateral hemisphere or ipsilateral rostral thalamus. The neurons were then stained by intracellular injection of horseradish peroxidase. The laminar distribution of these neurons was compared to that of cortical cells stained retrogradely after injection of wheat germ agglutinin/HRP in the ipsilateral or contralateral neostriatum. The morphological features of physiologically identified corticostriatal neurons, their laminar organization, and their responses to stimulation were examined and compared with crossed corticocortical and brainstem-projecting cells. Crossed corticostriatal cells of the medial agranular cortical field were medium-sized pyramidal neurons found in the superficial part of layer V and in the deep part of layer III. Their basilar dendritic fields and initial intracortical axon collateral arborizations were coextensive with the layer defined by the distribution of corticostriatal neurons. The apical dendrites were thin and sparsely branched but consistently reached layer I, where they made a small arborization. These morphological features were shared by cortical neurons projecting to contralateral neocortex but not responding antidromically to stimulation of contralateral neostriatum, but they were not shared by brainstem-projecting cortical cells. Orthodromic responses to contralateral cortical stimulation consisted of brief excitatory postsynaptic potentials that were followed by powerful and longer-lasting inhibitory postsynaptic potentials. Corticostriatal cells also exhibited small excitatory postsynaptic potentials in response to thalamic stimulation. Many crossed corticostriatal neurons were also commissural corticocortical neurons. The results of reciprocal collision tests showed that this was due to the existence of two separate axonal branches, one projecting to contralateral neocortex and one to contralateral neostriatum. Intracellular staining of these neurons revealed ipsilateral axonal projections to the neostriatum and cortex.     

Corticocortical connections of cat primary auditory cortex (AI): laminar organization and identification of supragranular neurons projecting to area AII

The laminar distribution and structure of the supragranular cells projecting from primary auditory cortex (AI) to the second auditory cortex (AII) in the cat were studied with horseradish peroxidase. Injections in AII retrogradely labeled somata in ipsilateral cortical layers I-VI of AI. A bimodal laminar disposition was found, with approximately 40% of the labeled cells in layer III, 25% in layer V, and 10-15% each in layers II, IV, and VI; only a few cells were found in layer I. The labeled cells were scattered in small aggregates between which unlabeled neurons were interspersed. There was some, though not a strict, topographical distribution of the labeled cells according to the locus of the injection in AII. Injections in the caudal part of AII labeled cells in more rostral AI, while rostral AII injections labeled cells in more caudal AI. Ventral AII injections labeled more ventrally located AI cells, while more dorsal AII injections labeled more dorsally situated AI cells. AII injections also labeled cells in other auditory cortex subdivisions, including the posterior ectosylvian, ventroposterior, temporal, and dorsal auditory zone/suprasylvian fringe cortical areas, and in some non-auditory cortical areas. In layers II and III, both pyramidal and non-pyramidal cells were labeled. More pyramidal cells were labeled in layer III than layer II (80% vs. 62%), and the proportion of non-pyramidal cells in layer II was more than twice that in layer IV (27% vs. 12%). The types of labeled cells were distinguished from one another on the basis of size, somatic and dendritic shape, and laminar distribution. The profiles of labeled cells in these experiments were compared to, and correlated with, those in Golgi-impregnated material. In layer II, the classes of corticocortical projecting cells consisted of small and medium-sized pyramidal, bipolar, and multipolar cells. Those in layer III included small, medium-sized, and large pyramidal neurons, and bipolar and multipolar cells. The average somatic area of the labeled cells did not differ significantly from that of the unlabeled cells, and both were about equal in somatic size to neurons accumulating tritiated gamma-aminobutyric acid in layers II and III. These findings suggest that there is convergent, ipsilateral input onto AII from every layer in AI, and from other cortical auditory and non-auditory areas. A morphologically heterogeneous population of cells in AI contributes to these projections. Diversity in the cytological origins of corticocortical projections implies functional differences between layers II and III since the latter also projects commissural, while layer II in the cat, does not.     

Size, laminar and columnar distribution of efferent cells in the sensory-motor cortex of monkeys.

The cells of origin of cortico-cortical and subcortical projections from the subfields of the somatic sensory area and from the motor cortex have been identified in cynomolgus and squirrel monkeys by the retrograde axonal transport method. The somata of the cells of origin of a particular fiber system have a specific laminar or sublaminar distribution. The somata of the majority of cortico-cortical cells lie in the supragranular layers. Those projecting to the opposite cortex are confined to the deeper half of layer III (layer IIIB). Ipsilateral cortico-cortical neurons lie mainly superficial to them in layers IIIA and II, but in the second somatic sensory area (SII) and in area 2 of the first (SI), small numbers are also found in layer V. Corticospinal cells lie in the deeper part of layer V and corticostriatal cells in the superficial part. Corticopontine, corticobulbar and corticorubral cells lie in between. The majority of corticothalamic cells lies in layer VI but a second, smaller population is found in the deep part of layer V. The cells giving rise to a particular set of efferent connections can be distinguished in terms of size and, with the exception of the corticospinal cells, their size does not vary greatly from area to area. In many cases, the size and laminar specificity indicates that cells sending axons to one site cannot have collateral branches projecting to another. In most of the fiber systems studied, labeled cells form single or multiple strips, 0.5–1 mm wide and oriented mediolaterally across the cortex. The strips appear in all of the subfields of the somatic sensory and motor areas and may form the basis of the clustering of like groups of efferent neurons demonstrable in physiological studies.     

Cells of origin and terminal distribution of corticostriatal fibers arising in the sensory-motor cortex of monkeys.

The cells of origin of the corticostriatal projection have been identified in squirrel monkeys by the use of the retrograde horseradish peroxidase method. In the subfields of the somatic sensory, motor, parietal and frontal areas of the cortex, cells projecting to the ipsilateral striatum are relatively sparsely distributed and form a group of small- to medium-sized pyramidal cells with an average somal diameter from area to area of 14-16 mum. Such cells are found only in layer V of the cortex (mainly in the more superficial parts of the layer). Since they are consistently smaller than the pyramidal cells of layer V that project to the brainstem and spinal cord and since they lie outside layer VI which gives rise to corticothalamic axons, the corticostriatal axons are unlikely to be collaterals of axons projecting to other sites. The cells of origin of the crossed corticostriatal projection are also found in layer V and are pyramidal cells with somal diameters in the same range as above. They are found only in areas 4, 8, and 6. Studies with the anterograde, autoradiographic method in rhesus, cynomologous and squirrel monkeys, indicate that the somatic sensory areas project to most of the antero-posterior extent of the ipsilateral putamen. Subareas 3a, 3b, 1 and 2 of the somatic sensory cortex project to the same region and the projection overlaps similarly extensive projections from the motor and certain other areas of the cortex. However, in each case the pattern of terminal labeling is in the form of interrupted clusters, strips and bands. A single small injection of the cortex is associated with only one or two such clusters of terminal labeling. This seems to imply that individual corticostriatal fibers end in a very restricted manner and that the terminal ramifications of fibers from one cortical area may alternate in the putamen with those arising in other areas.     

Quantitative analyses of neurons projecting to primary motor cortex zones controlling limb movements in the rat

The objective was to determine if projections of single neurons to primary motor cortex preferentially terminate in several efferent zones that could form synergies for the execution of limb movements. Intracortical microstimulation was used to identify zones evoking hip flexion (HF), elbow flexion (EF), and both plantarflexion (PF) and dorsiflexion (DF) about the ankle. Histological examination showed that the zones from which some movements were evoked extended beyond the agranular cortex into granular cortex. Fluorogold, Fast blue, and propridium iodide or rhodamine-labeled dextran were injected into three of these four efferent zones in each rat. There was a virtual absence of multiple-labeled cells despite having an intermingling of different-colored cells of which 15% in frontal cortex were less than 1.2 mm away from a neighboring neuron that projected to a different efferent zone. This suggests that single neurons projecting to the motor cortex do not hard-wire specific synergies but rather project to single efferent zones in order to offer the greatest degree of freedom for the generation of movements. The distribution of ventral posterolateral and ventrolateral thalamic nucleus labeling depended on whether the injections were in granular or agranular cortex. Conversely, frontal cortex projections to motor efferent zones were made irrespective of their location in either granular or agranular cortex and thereby supporting their presumed role in the control of movements. Hindlimb motor cortex injections yielded retrograde labeling that extended into the more localised distribution of frontal cortex neurons retrogradely labeled from forelimb injections. This may allow hindlimb movements to be synchronized by forelimb movements during walking on challenging terrain.     

Overlapping corticostriatal projections from the supplementary motor area and the primary motor cortex in the macaque monkey: An anterograde double labeling study

The purpose of the present study was to determine if the cortical efferents from homologous body regions of the supplementary motor area (SMA) and the primary motor cortex (MI) project to separate or to overlapping regions in the striatum. In order to investigate the dual corticostriatal projections, we employed an anterograde double labeling paradigm in which two tracers could be simultaneously detected in the same histological section. Prior to the injections, the forelimb representation in the two cortical motor areas was identified by using intracortical microstimulation in four Japanese monkeys (Macaca fuscata). Multiple injections of biotinylated dextran amine (BDA) were made into the forelimb regions of MI and wheat germ agglutinin conjugated horseradish peroxidase (WGA-HRP) was injected into the arm region of the SMA. In additional animals, the tracers were reversed such that BDA was injected into the SMA and WGA-HRP was injected into the MI. The tissue was processed sequentially using different chromogens in order to visualize both tracers in a single section. We analyzed the distribution of the ipsilateral anterograde label. The striatal labeling from each cortical area basically consisted of a wide band of patchy dense labeling interrupted by lighter labeling. The SMA striatal projections were located mainly within the putamen, distributing from the level of the anterior commissure to the most posterior extent of the putamen. At an intermediate level, the label spread obliquely from the ventrolateral edge of the putamen dorsomedially as far as the lateral edge of the caudate nucleus. The label from the MI was observed in comparable portions of the putamen, although the SMA projections were shifted more anterior and dorsomedial to the MI projections and the heaviest projections from the SMA and the MI were separately located. On the basis of the double anterograde labeling technique, we found considerable overlap mainly in the central portion of the putamen from the SMA and MI forelimb representation. These results suggest that the homologous body regions of the SMA and MI send widespread, and substantially overlapping projections, to portions of the striatum. © 1996 Wiley-Liss, Inc.     

Cortical Regulation of Striatal Neuropeptide Y (NPY)-Containing Neurons in the Rat

This study examined the functional relationships established by nigral, cortical, and thalamic striatal afferent pathways with neuropeptide Y (NPY)-containing neurons in the rat rostral striatum by coupling selective deafferentation procedures and NPY immunohistochemistry. Previous experiments have shown that after unilateral 6-hydroxydopamine (6-OHDA)-induced degeneration of nigrostriatal dopaminergic neurons, the mean number of NPY-immunoreactive (Ir) neurons per frontal section was increased in the striatum ipsilateral to the lesion side and unaltered in the contralateral striatum. The present topographical analysis of the 6-OHDA lesion effects led us to state that the increase in NPY-Ir neuron density occurs in restricted ventral and medial zones of the ipsilateral striatum. Unilateral ablation of the frontoparietal cerebral cortex by thermocoagulation was moreover shown to elicit, 20 - 30 days later, a significant bilateral increase in the number of striatal NPY-Ir cells. The increase was more marked in the striatum ipsilateral to the hemidecortication where it was similar in amplitude to that induced by the 6-OHDA lesion. The topographical analysis of the cortical lesion effects also revealed an uneven striatal response, but, in contrast to that observed for the 6-OHDA lesion, changes were restricted to dorsolateral areas of the striatum in both brain sides, revealing an apparent complementarity of nigral dopaminergic and cortical influences over striatal NPY neuronal system. Combined unilateral nigral and cortical lesions surprisingly counteracted in a survival time dependent manner the effects of each lesion considered separately. In that condition topographical changes related to the 6-OHDA lesion totally disappeared and those related to the cortical lesion were attenuated but still present. These results suggest that expression of striatal dopamine - NPY interaction is dependent on corticostriatal transmission. Interestingly lesion of thalamic areas projecting to the striatum did not significantly modify the mean number of NPY-Ir neurons determined per section from the whole striatal surface, but selectively increased the NPY neuron density in the mediodorsal region of the striatum, suggesting that the striatal NPY-containing neuronal system is also influenced by thalamostriatal projections.