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Distribution of calmodulin, calbindin-D28k and calretinin among rat olfactory nerve bundles.

Calmodulin, calbindin-D28k and calretinin are calcium-binding proteins largely distributed in the bipolar olfactory receptor cells. In the olfactory epithelium their distribution seemed to be random. Using immunohistochemistry we have examined their localization in rat olfactory axons extending to the olfactory bulb. Sections were analyzed both horizontally and vertically. Almost all fibers were immunoreactive for one of the three intracellular calcium-binding proteins whose distribution was not random among the bundles. Three different subclasses of fibers could be detected: calbindin-D28k and calretinin immunoreactivities were restricted to external fibers whereas calmodulin immunoreactivity was intense, abundant and largely distributed throughout the internal portion of the olfactory nerve. This additional degree of organization detected in the olfactory axons might play a role in odor discrimination.

Animals↗

Immunohistochemical localization of calcium-binding proteins in the human cutaneous sensory corpuscles.

The localization of the calcium-binding proteins (CaBP) calbindin-D28k (CB), parvalbumin (PV) and S-100 protein (S100P) in the human cutaneous sensory corpuscles was studied by immunohistochemical procedure using monoclonal antibodies. Occurrence of CB, PV and S100P immunoreactivity (IR) was observed in the lamellar cells of Meissner's corpuscles. In the pacinian corpuscles, S100P IR was restricted to the inner-core cells whereas CB and PV IR were found labelling the axon, inner core, outer core and capsule. At the light-microscope level of resolution, the presence of IR in the axon of Meissner's corpuscles cannot be ensured. Since calcium ions (Ca2+) seem to participate in the mechanoreceptor electrogenesis, present results suggest that CaBP could be involved in buffering and/or transport of Ca2+ within the specialized cells surrounding the axon tips of sensory corpuscles, thus, maintaining the periaxonal microenvironment.

Adult↗

Immunohistochemical localization of calbindin-D28K in telencephalic regions of microencephalic rats.

The localization of calbindin neurons was studied in different brain areas of rats rendered microencephalic by gestational methylazoxymethanol acetate (MAM) treatment. In layers VI and V of the cortex, the only recognizable layers in MAM-treated rats, a higher density of calbindin interneurons and an apparent increase in protein expression was observed. In the hippocampus, calbindin pattern was essentially preserved, despite the dramatic decrease in size. In other telencephalic regions, calbindin distribution was not changed except for the septum, where a large increase of calbindin neurons was observed. The present results suggest that the MAM model may be used to investigate the role of calbindin.

Animals↗

Calbindin D28k-like immunoreactivity in the gustatory epithelium in the rat.

The distribution of calbindin D28k (CB)-like immunoreactivity (-LI) in the gustatory epithelium was examined in the adult rat. In the circumvallate and foliate papillae, CB-like immunoreactive (-IR) nerve fibers were observed in the subgemmal region, and some of these penetrated the taste buds. Two or three spindle-shaped gustatory cells displayed CB-LI in each taste bud of these lingual papillae; the immunoreactivity was restricted to the cytoplasm. In the fungiform papilla, CB-IR nerve fibers were detected in the subgemmal region, but no CB-IR cells were observed in the taste buds of the fungiform papillae. In the taste buds of the incisive papillae, many CB-IR intragemmal nerve fibers were observed, but no apparent CB-IR cells were detected. In the soft palate, CB-IR nerve fibers associated with the taste buds were also observed, but no CB-IR cells were detected in the taste buds. The present findings indicate that CB-IR gustatory cells were only localized in the taste buds in the posterior lingual papillae (circumvallate and foliate papillae), but not in the taste buds in other gustatory epithelium.

Animals↗

Analysis of parvalbumin and calbindin D28k-immunoreactive neurons in dorsal root ganglia of rat in relation to their cytochrome oxidase and carbonic anhydrase content.

Histochemical and immunohistochemical techniques were used to determine relationships between the parvalbumin or calbindin D28k content and the cytochrome oxidase or carbonic anhydrase activity of neurons in lumbar dorsal root ganglia in rat. Subpopulations of dorsal root ganglion neurons that displayed parvalbumin- or calbindin D28k-immunoreactivity were classified as containing either light, moderate or dense histochemical reaction product for cytochrome oxidase and either a positive or negative reaction for carbonic anhydrase. It was found that approximately 90% of all parvalbumin and calbindin D28k-immunoreactive cells exhibited dense staining for cytochrome oxidase and that 87% of parvalbumin- and 76% of calbindin D28k-immunoreactive cells were positive for carbonic anhydrase. Conversely, 85% of all cells with a dense cytochrome oxidase reaction contained parvalbumin and calbindin D28k. Although not quantified, it appeared that many, but not all, carbonic anhydrase-positive cells contained parvalbumin or calbindin D28k. These results indicate the existence of a subpopulation of primary sensory neurons that contains parvalbumin and calbindin D28k and that expresses high levels of cytochrome oxidase and carbonic anhydrase activity. It is suggested that primary afferent neurons with this cytochemical profile transmit a sensory modality that requires them to discharge rapidly and/or frequently. The existence of a subpopulation of carbonic anhydrase-positive cells that lack immunoreactivity for parvalbumin or calbindin D28k suggests that the role of carbonic anhydrase in some sensory neurons is unrelated to functions requiring these calcium binding proteins.

Animals↗

Septal GABAergic neurons innervate inhibitory interneurons in the hippocampus of the macaque monkey.

The septohippocampal projection was visualized in three Macaca mulatta monkeys by anterograde transport of Phaseolus vulgaris leucoagglutinin. Following injections of the lectin into the medial septal nucleus, P. vulgaris leucoagglutinin-labelled fibres were found in the hippocampal complex, mainly in stratum oriens of the CA1 subfield, throughout the CA3 subfield, and in the hilus and stratum moleculare of the dentate gyrus. The majority of labelled axons were varicose, and formed multiple contacts with cell bodies and dendrites of calbindin D28k- and parvalbumin-immunoreactive non-pyramidal cells. GABA immunoreactivity of P. vulgaris leucoagglutinin-labelled axons and their postsynaptic targets was investigated by sectioning varicose axon segments for correlated light and electron microscopy, and processing alternate ultrathin sections for postembedding immunogold staining for GABA. All P. vulgaris leucoagglutinin-labelled boutons examined were GABA-immunoreactive and the majority of them formed symmetrical synapses with GABA-immunoreactive cell bodies and dendrites. The results demonstrate that a GABAergic septohippocampal pathway exists in the monkey, and, similar to the rat, terminates on different types of GABAergic neurons, including the parvalbumin- and calbindin D28k-containing non-pyramidal cells.

Animals↗

Calretinin in rat brain: an immunohistochemical study.

Calretinin is a calcium-binding protein related to calbindin-D28k; both are present in different though overlapping sets of neurons in brains of birds and mammals. We describe in detail the pattern of calretinin immunoreactivity in the rat brain. As in chick brain, calretinin immunoreactivity is abundant in various sensory pathways (particularly certain cells and fibres of the cochlear nuclei and olfactory bulb), in the heterogeneous parts of the brainstem and in parts of the hypothalamus. Many primary sensory fibres are strongly positive. Major groups of calretinin-positive neurons also include the thalamic reticular nucleus, triangular septal nucleus, lateral mammillary nucleus and substantia nigra pars compacta. Many other calretinin-positive cells are recognizable as local inhibitory neurons. Calretinin is absent from all but a few cells in the cerebral cortex, and is never found in motor neurons. There are also some distinctive positive structures whose identity is uncertain, notably irregular "shells" of cells and fibres around the thalamus and in the amygdala and an unnamed cell type in the vestibulocerebellum.

Animals↗

Calretinin is present in non-pyramidal cells of the rat hippocampus--II. Co-existence with other calcium binding proteins and GABA.

The possible co-existence of calretinin with other calcium binding proteins, parvalbumin and calbindin D28k, and with GABA, was studied in non-pyramidal cells of the rat dorsal hippocampal formation, using the mirror technique. The majority of the calretinin-containing neurons (83%) were found to be immunoreactive for GABA (79% in the dentate gyrus, 84% in the CA2-3, and 88% in the CA1 subfield). Most of the GABA-negative calretinin-immunoreactive neurons were located in the hilus of the dentate gyrus and in stratum lucidum of the CA3 subfield. Detailed analysis of the calretinin-immunoreactive cells of these subfields revealed that the two morphologically distinct types of calretinin neurons, i.e. the spiny and the spine-free cells, differ in their immunoreactivity for GABA. The overwhelming majority (92%) of the spine-free neurons were GABA-positive, whereas the immunoreactivity of spiny cells was ambiguous. At the sensitivity threshold of the immunocytochemical techniques used in the present study, most of the spiny cells (89%) had to be considered as GABA-negative, although the staining intensity in their cell bodies was somewhat above background level. Colchicine treatment resulted in a degeneration of calretinin-immunoreactive neurons; therefore, its effect on the GABA content of spiny neurons could not be evaluated. Nevertheless, the observations suggest that calretinin-containing neurons are heterogeneous both morphologically and neurochemically. Examination of the co-existence of calcium binding proteins revealed that none of the hippocampal cells contained both calretinin and parvalbumin in any regions of the hippocampal formation. Some overlap was detected between the calretinin- and the calbindin D28k-containing cell populations, 5.1% of the former and 6.2% of the latter were immunoreactive for both calcium binding proteins. This may be due to a small degree of cross-reactivity of the calbindin D28k antiserum with calretinin. Thus, our results demonstrate that the majority of calretinin-immunoreactive neurons are GABAergic and represent a subpopulation of non-pyramidal cells with no or only a negligible overlap with the subpopulations containing the other calcium binding proteins, parvalbumin and calbindin.

Animals↗

Calbindin D28k-containing nonpyramidal cells in the rat hippocampus: their immunoreactivity for GABA and projection to the medial septum.

Calbindin D28k-containing non-pyramidal cells were found in all layers and subfields of the hippocampus, with the highest frequency in stratum radiatum of the CA1-CA3 subfields. A large number of these neurons had a vertically oriented dendritic tree, often restricted to to stratum radiatum. In stratum oriens and near to the border of strata radiatum and lacunosum moleculare cells with horizontally running dendrites were also found. Multipolar cells were most common in stratum radiatum of the CA3 region. The GABAergic nature of the calbindin D28k-containing non-pyramidal cells was studied using the "mirror" technique. Adjacent thick sections were immunostained for calbindin D28k and GABA, and halved neurons were identified on the common surfaces. The majority of calbindin D28k-containing non-pyramidal cells were shown to be GABAergic. The GABA-negative calbindin cells were found in relatively large numbers in stratum oriens of the CA1-CA3 region, and occasionally in strata radiatum and pyramidale of CA2, and in stratum radiatum of the CA3c region near to the border of the dentate hilus. However, even in these cells a weak immunostaining, only slightly but consistently above background level, was always observed. Earlier studies have demonstrated that the somata of GABAergic neurons with distant projections may contain a level of GABA that is below the detection threshold of immunocytochemistry. Here we provide direct evidence that the calbindin-containing non-pyramidal cells were among those projecting to the medial septum. Following horseradish peroxidase injections into the medial septum 80% of the retrogradely labelled non-pyramidal cells were found to be immunoreactive for calbindin D28k, and 20% contained neuropeptide Y. These results suggest that the calbindin D28k-containing and apparently GABA-immunonegative non-pyramidal cells in stratum oriens of the CA1-CA3 regions may also be GABAergic, but have a distant projection, that is, to the medial septum.

Animals↗

Calretinin and calbindin-D28k in rat brain: patterns of partial co-localization.

Calretinin and calbindin-D28k are homologous calcium-binding proteins, each present in a variety of neurons in the brain. Their distributions in the rat brain have been compared at the cellular level to determine whether they tend to occur in the same or in different cells, and to determine whether calbindin-positive cells show any common features once crossreaction with calretinin has been eliminated. The results show great heterogeneity. Most cells which contain one of the proteins do not contain the other, but many cells do contain both; even in the ventral cochlear nucleus, where there is abundant calretinin and most calbindin-like immunoreactivity is due to crossreaction, a few cells contain both proteins. In the substantia nigra and ventral tegmental area, many cells are double-positive but some only contain one or the other protein. Only the triangular septal nucleus is uniformly positive for both proteins. Cells which look like local-circuit neurons in many forebrain areas (cortex, hippocampus, olfactory bulb, anterior olfactory nucleus) are exclusively positive for either calretinin or calbindin, in spite of their similar morphology. In the more heterogeneous parts of the brain (including hypothalamus central gray and substantia gelatinosa), there are mixtures of calretinin-positive, calbindin-positive, and double-positive cells. In comparison with previous data on the chick, some aspects of the distributions are conserved, but double-positive cells are more frequent in the rat. The degree of heterogeneity observed, even within comparatively well-defined neuronal populations, makes it difficult to infer in what neuronal properties these proteins could be involved.

Animals↗

Mouse fetal trisomy 13 and hypotrophy of the spinal cord: effect on calbindin-D28k and calretinin expressed by neurons of the spinal cord and dorsal root ganglia.

Trisomy 13 was detected in 10% of mouse embryos obtained from pregnant females which were doubly heterozygous for Robertsonian chromosomes involving chromosome 13. The developing dorsal root ganglia and spinal cords were examined in trisomy 13 and littermate control mice between days 12 and 18 of gestation (E12-18). The overall size of the dorsal root ganglia and number of ganglion cells within a given ganglion were not altered, but the number of neurons immunoreactive for calbindin and calretinin was reduced. The trisomic spinal cord was reduced in size with neurons lying in a tightly compact distribution in the gray matter. In trisomic fetuses, the extent of the neuropil of the spinal cord was reduced, and may represent a diminished field of interneuronal connectivity, due to reduced arborization of dendritic processes of the neurons present, particularly of calbindin-immunostained neurons. Furthermore, the subpopulation of calretinin-immunoreactive neurons and axons was also reduced in developing trisomic gray and white matter, respectively. Thus, overexpression of genes on mouse chromosome 13 exerts a deleterious effect on the development of neuropil, affecting both dendritic and axonal arborization in the trisomy 13 mouse. The defect of calbindin or calretinin expression by subsets of dorsal root ganglion or spinal cord neurons may result from deficient cell-to-cell interactions with targets which are hypoplastic.

Animals↗

Differential localization of NADPH-diaphorase and calbindin-D28k within the cholinergic neurons of the basal forebrain, striatum and brainstem in the rat, monkey, baboon and human.

The localization of Calbindin-D28k and NADPH-diaphorase in the cholinergic neurons of the basal forebrain, striatum and brainstem was investigated in the rat, monkey, baboon and human using calbindin and choline acetyltransferase immunohistochemistry and NADPH-diaphorase histochemistry. Considerable regional and species-specific variations were observed. Double-stained sections demonstrated that NADPH-diaphorase activity occurred in as much as 20-30% of basal forebrain cholinergic neurons in the rat but in virtually none of those neurons in the monkey, baboon or human. In all of the species studied, virtually every cholinergic neuron within the pedunculopontine and laterodorsal tegmental nuclei contained NADPH-diaphorase activity, while none of the cholinergic neurons of the striatum did so. In the rat brain, calbindin immunoreactivity was not present in any of the cholinergic neurons of the basal forebrain, while in the primate brain virtually all of the basal forebrain cholinergic neurons were also calbindin-positive. None of the cholinergic neurons of the striatum, pedunculopontine nucleus or laterodorsal tegmental nucleus were found to be calbindin-positive in any of the species examined. These results demonstrate major species-specific differences in the cytochemical signatures of the basal forebrain cholinergic neurons, in contrast to the cholinergic neurons of the striatum and brainstem, which displayed little interspecies variation with respect to the markers that were used in this study. Our findings also suggest that caution must be exercised in using results from studies of rodent basal forebrain cholinergic systems to infer the role of this system in the primate brain.

Animals↗

Relative loss of the striatal striosome compartment, defined by calbindin-D28k immunostaining, following developmental hypoxic-ischemic injury.

The striatum is especially vulnerable to hypoxic-ischemic injury, both in adulthood and during development. Striatal injury is likely to play a major role in the chronic abnormalities of motor control which occur as a consequence of developmental hypoxia-ischemia. Previous studies have shown that two striatal neuron phenotypes, cholinergic and NADPH-diaphorase-positive, are resistant to developmental hypoxia-ischemia, but little is otherwise known of patterns of vulnerability among other striatal neurons. In particular, there has been no data available about patterns of vulnerability within the major striatal neuron group, the medium-sized neurons. Since a major anatomical and functional organization of these neurons is in their localization to either the striosome or the matrix compartments, we have examined the effect of developmental hypoxia-ischemia on these compartments using a quantitative morphologic analysis of immunostaining for the calcium-binding protein calbindin-D28k. We have found that there is a predominant loss of the striosome compartment; in the presence of a mean loss of 33% of total striatal area, there was a 49% decrease in striosomal area. There was also a 41% reduction in the number of striosomes, and a small (14%) but significant decrease in the mean area of individual striosomes. The striosome loss was uniform in the rostrocaudal dimension. At a cellular level, the density of calbindin-positive neurons, expressed as number per unit area, was preserved. While there are several possible explanations for the selective loss of the striosome compartment, one hypothesis is that the lower level of calbindin within these neurons makes them more vulnerable to increases in intracellular calcium, which has been postulated to play a role in hypoxic-ischemic injury. The predominant loss of the striosome compartment following hypoxic-ischemic injury may lead to an imbalance with the functionally distinct matrix system. Such an imbalance may contribute to the abnormalities of motor control observed after this form of injury.

Age Factors↗

Immunocytochemical properties of rat renal afferent neurons in dorsal root ganglia: a quantitative study.

Immunocytochemical properties of dorsal root ganglion neurons innervating the kidney were studied with retrograde tracing of Fluorogold or Fast Blue dyes applied to the cut renal nerves in the rat. The proportions and sizes of renal afferent neurons labelled with a variety of markers were quantified in T9-L1 dorsal root ganglia from five rats. Compared with the overall size distribution in these ganglia, renal afferent neurons were mainly small with a few medium-sized neurons. The majority (79%) of renal afferent dorsal root ganglion neuronal somata were unlabelled by an anti-neurofilament antibody, RT79, and classified as neurofilament-poor with probable C-fibres. These had an approximately normal distribution of cell sizes. Only 21% were RT79-positive and classified as neurofilament-rich with probable A-fibres, and even these were small to medium sized cells, consistent with them being mostly A delta-fibre neurons. Percentages of renal afferent neurons showing labelling were as follows: peripherin-like immunoreactivity: 69%; calcitonin-gene related peptide: 93%; substance P: 37%; the lectins soybean agglutinin: 57% and peanut agglutinin: 68%; Calbindin D28k-like immunoreactivity: 21% (only weak labelling); carbonic anhydrase like immunoreactivity: 0%. There were differences between probable C-fibre and probable A-fibre neurons, classified according to their labelling with RT97. The percentages of RT97-negative and RT97-positive neurons respectively labelled with the other markers were as follows: peripherin-like immunoreactivity: 82%, 25%; calcitonin gene-related peptide-like immunoreactivity: 99%, 79%; substance P-like immunoreactivity: 43%, 0%; soybean agglutinin: 69%, 24%; peanut agglutinin: 76%, 47%; calbindin-like immunoreactivity: 26%, 0%. Thus, the biggest differences between the probable A- and C-fibre renal afferent neurons were in their peripherin, substance P and calbindin contents. Thus, renal afferent neurons in the dorsal root ganglion are not homogeneous and it is suggested the differences may relate to the known different afferent receptor types within the kidney. It is suggested that the low proportion of neurons with substance P-like immunoreactivity in the renal afferent innervation compared to that of other viscera may relate to the role of the renal vasculature in urine formation.

Animals↗

Sparing of the dopaminergic neurons containing calbindin-D28k and of the dopaminergic mesocortical projections in weaver mutant mice.

In mice carrying the weaver mutation there is a spontaneous degeneration of dopaminergic neurons that is heterogeneous among cell groups: nigrostriatal neurons are more affected than mesolimbic neurons, while involvement of the mesocortical system is controversial. We questioned whether the pattern of cell loss in mesencephalon and fiber depletion in telencephalon could be related to the differential content of Calbindin-D28k in dopaminergic cells. The mesencephalon of seven-month-old mutants was serially sectioned and alternate series were immunostained with tyrosine hydroxylase and Calbindin-D28k. Cell counts indicated a 40% loss for the ensemble of dopamine mesencephalic neurons. However, double-immunostained preparations revealed that this cell loss was restricted to the neurons that lacked Calbindin-D28k, which were reduced by 72%, while the dopaminergic neurons containing Calbindin-D28k were completely spared. Calbindin-D28k was present in both the cytoplasm and nucleus of the dopaminergic cells. This nuclear localization was confirmed at the ultrastructural level. In the telencephalon of weaver mutants, areas receiving projections from the Calbindin-D28k-positive dopaminergic neurons, such as the cerebral cortex, contained normal densities of fibers, while areas harboring projections from the non-Calbindin-D28k dopaminergic neurons, such as the dorsal striatum, had reduced amounts of fibers. The vulnerability pattern in the mesencephalon of weaver mutants bears similarities to that described in idiopathic Parkinson's disease or in N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced Parkinsonism: Calbindin-D28k may thus delimit a group of dopaminergic neurons resistant to cell death in different conditions. On the other hand, the vulnerability pattern of dopaminergic fibers in weaver differs from that of Parkinson's disease, since there is a complete sparing of the dopaminergic mesocortical projection in weaver, contrasting with the damage of these projections in Parkinson's disease.

Afferent Pathways↗

Differential distribution of calbindin-D28k and parvalbumin in somatic and visceral sensory neurons.

The purpose of the present investigation was to determine whether calbindin-D28k and parvalbumin are distributed to different subpopulations of somatic and visceral sensory neurons. Immunofluorescent and retrograde techniques were combined to examine the distribution of calbindin- and parvalbumin-like immunoreactivity in the cell bodies of somatic and visceral primary afferent neurons in dorsal root ganglia L1-S1 of rats. Calbindin and parvalbumin were differentially distributed to essentially non-overlapping subpopulations of primary sensory neurons that could be distinguished by their segmental and size distributions, as well as by their innervation of somatic and visceral structures. Calbindin-like immunoreactivity was found in a population of smaller-sized cell bodies comprising approximately 14%of all dorsal root ganglion cells examined, with the proportions being greatest in L6 and S1. In contrast, parvalbumin was found in a population of larger-sized cells that made up about 11% of dorsal root ganglion cells and that were most concentrated in L4 and L5. Sensory neurons were further characterized by retrograde transport following the application of the neuroanatomical tracer FluoroGold to somatic (sural and gastrocnemius) and visceral (hypogastric and pelvic) nerves. Somatic tissues were innervated by a population of calbindin-containing as well as a separate population of parvalbumin-containing sensory neurons. In contrast, afferent neurons innervating visceral structures contained only a subpopulation of calbindin-containing neurons and very few parvalbumin-positive cells.

Animals↗

Ultrastructural localization of calbindin-D28k and GABA in the matrix compartment of the rat caudate-putamen nuclei.

The calcium binding protein, Calbindin-D28k, is known to be localized within spiny neurons of the matrix of the dorsal striatum, caudate-putamen nuclei. This compartment is also known to contain an abundance of GABAergic neurons and to receive extensive input from excitatory limbic and cortical afferents whose activation produces rapid influxes of calcium in neuronal targets. We used electron microscopic immunocytochemistry to examine a potential role for calbindin in GABAergic neurons in the caudate-putamen nuclei. Sections of striatal tissue from acrolein-fixed adult rat brains were dual-labeled using immunoperoxidase for the localization of rabbit anti-calbindin and immunogold-silver for the localization of rat anti-GABA antibodies. Calbindin-D28k and GABA were mainly co-localized in somata and large dendrites. The peroxidase reaction product for calbindin was diffusely distributed throughout the neuronal cytoplasm, but appeared more densely localized along asymmetric, excitatory-type, postsynaptic junctions of dendritic spines, as well as saccules of smooth endoplasmic reticulum near dendritic appositions. In contrast, the immunogold-silver labeling for GABA was largely restricted to perikarya and large dendrites. Axon terminals forming symmetric junctions were also sometimes dual-labeled for calbindin and GABA. However, the majority of the calbindin-immunoreactive terminals did not contain GABA and many formed asymmetric excitatory-type synapses with either unlabeled or calbindin-labeled dendritic spines. These results suggest that, in the striatal matrix, Calbindin-D28k contributes to the immobilization of calcium (i) in selectively activated postsynaptic spines of GABAergic and possibly non-GABAergic neurons and (ii) in terminals containing GABA as well as other excitatory and inhibitory transmitters. The extent to which calbindin is able to restrict the cytosolic increases in calcium to selective sites of utilization in these neurons may have important consequences for normal synaptic function and for neuroprotection against excitoxicity.

Animals↗

Patterns of overlap and segregation between insular cortical, intermediodorsal thalamic and basal amygdaloid afferents in the nucleus accumbens of the rat.

Regions of the prefrontal cortex that project to the nucleus accumbens in the rat receive input from midline thalamic and basal amygdaloid nuclei which also project to the same striatal region as their prefrontal cortical target. For example, the prelimbic cortex projects to the medial nucleus accumbens, and receives input from the paraventricular thalamic nucleus and the parvicellular basal amygdala. These latter two areas also project to the medial nucleus accumbens. It has been shown that afferents from the prelimbic cortex, the paraventricular thalamic nucleus and the parvicellular basal amygdala to the nucleus accumbens overlap or are separated in the nucleus accumbens, depending upon their position in the shell and core. The dorsal agranular insular cortex, the intermediodorsal thalamic nucleus and the magnocellular basal amygdaloid nucleus terminate in the lateral part of the nucleus accumbens and adjacent ventral part of the caudate-putamen. The intermediodorsal thalamic nucleus and the magnocellular basal amygdaloid nucleus reach both the dorsal agranular insular cortex and the lateral nucleus accumbens, and thus appear positioned to influence the prefrontal corticostriatal system at cortical and striatal levels. However, all three afferent systems have a heterogeneous distribution within this striatal region, and whether these projections actually reach the same areas is unknown. We investigated the patterns of separation and overlap in the nucleus accumbens between dorsal agranular insular cortical, magnocellular basal amygdaloid and intermediodorsal thalamic afferents with respect to the histochemical features of the nucleus. Techniques allowing the detection of two different anterograde tracers, or a single anterograde tracer and Calbindin-D28k immunoreactivity, in the same tissue sections were used. The results demonstrate that the afferents from the dorsal agranular insular area and the intermediodorsal thalamic nucleus avoid the shell of the lateral nucleus accumbens, which receives strong inputs from the magnocellular basal amygdala. In the matrix of the core and the ventral part of the caudate-putamen, fibers from the superficial layers of the dorsal agranular insular area overlap precisely with afferents from the intermediodorsal nucleus. In the patches, projections from the deep layers of the dorsal agranular insular cortex coincide with those from the magnocellular basal amygdala. The present findings have implications for the compartmental structure of the nucleus accumbens and provide novel insights into the organizational principles of prefrontal corticostriatal circuits.

Afferent Pathways↗