Highlighting type A RRs as potential regulators of the dkHK1 multi-step phosphorelay pathway in Populus.

In previous studies, we highlighted a multistep phosphorelay (MSP) system in poplars composed of two hybrid-type Histidine aspartate Kinases, dkHK1a and dkHK1b, which interact with three Histidine Phosphotransfer proteins, dkHPt2, 7, and 9, which in turn interact with six type B Response Regulators. These interactions correspond to the dkHK1a-b/dkHPts/dkRRBs MSP. This MSP is putatively involved in an osmosensing pathway, as dkHK1a-b are orthologous to the Arabidopsis osmosensor AHK1, and able to complement a mutant yeast deleted for its osmosensors. Since type A RRs have been characterized as negative regulators in cytokinin MSP signaling due to their interaction with HPt proteins, we decided in this study to characterize poplar type A RRs and their implication in the MSP. For a global view of this MSP, we isolated 10 poplar type A RR cDNAs, and determined their subcellular localization to check the in silico prediction experimentally. For most of them, the in planta subcellular localization was as predicted, except for three RRAs, for which this experimental approach gave a more precise localization. Interaction studies using yeast two-hybrid and in planta BiFC assays, together with transcript expression analysis in poplar organs led to eight dkRRAs being singled out as partners which could interfere the dkHK1a-b/dkHPts/dkRRBs MSP identified in previous studies. Consequently, the results obtained in this study now provide an exhaustive view of dkHK1a-b partners belonging to a poplar MSP.

Medial prefrontal cortical innervation of the intercalated nuclear region of the amygdala.

The projections of the infralimbic area (IL) of the medial prefrontal cortex to the intercalated nuclei (ICNs) of the amygdala are thought to form a critical component of the forebrain circuitry for fear extinction. Despite the importance of these projections, there have been no focussed anatomical studies that have investigated the extent of IL inputs to different portions of the ICN complex. The present investigation used anterograde tract tracing in the rat to study the projections of the ventromedial PFC, including the IL, to the ICNs and surrounding amygdalar regions. Immunohistochemistry for the µ-opioid receptor (MOR) was used to identify the ICNs. At rostral levels of the amygdala there was a very dense projection to a far lateral portion of the capsular subdivision of the central nucleus (CLC) located between the main and medial ICNs, but only very light projections to these ICNs and the lateral ICNs. This distinct portion of the CLC receiving strong IL inputs was termed the capsular infralimbic target zone (CITZ), and was MOR-negative. Likewise, at more caudal levels of the amygdala, IL projections to the medial, lateral, and dorsal ICNs were light to moderate compared with projections to adjacent portions of the basolateral amygdala and amygdalostriatal transitional area. These findings suggest that the putative role of the IL-to-ICN connection in fear inhibition may be mediated by light to moderate projections from the IL to the medial ICN, and that the CITZ may be an equally important amygdalar target for this function.

Neuronal localization of M2 muscarinic receptor immunoreactivity in the rat amygdala.

Muscarinic cholinergic neurotransmission in the amygdala is critical for memory consolidation in emotional/motivational learning tasks, but little is known about the neuronal distribution of different receptor subtypes. Immunohistochemistry was used in the present investigation to localize the m2 receptor (M2R). Differential patterns of M2R-immunoreactivity (M2R-ir) were observed in the somata and neuropil of the various amygdalar nuclei. Neuropilar M2R-ir was strongest in rostral portions of the basolateral nuclear complex (BLC). M2R-positive (M2R+) somata were seen in low numbers in all nuclei of the amygdala. Most M2R+ neurons associated with the BLC were in the lateral nucleus and external capsule. These cells were nonpyramidal neurons that contained glutamatic acid decarboxylase (GAD), somatostatin (SOM), and neuropeptide Y (NPY), but not parvalbumin (PV), calretinin (CR), or cholecystokinin (CCK). Little or no M2R-ir was observed in GAD+, PV+, CR+, or CCK+ axons in the BLC, but it was seen in some SOM+ axons and many NPY+ axons. M2R-ir was found in a small number of spiny and aspiny neurons of the central nucleus that were mainly located along the lateral and ventral borders of its lateral subdivision. Many of these cells contained SOM and NPY. M2R+ neurons were also seen in the medial nucleus, including a distinct subpopulation of neurons that surrounded its anteroventral subdivision. The latter neurons were negative for all neuronal markers analyzed. The intercalated nuclei (INs) were associated with two types of large M2R+ neurons, spiny and aspiny. The small principal neurons of the INs were M2R-negative. The somata and dendrites of the large spiny neurons, which were actually found in a zone located just outside of the rostral INs, expressed SOM and NPY, but not GAD. These findings indicate that acetylcholine can modulate a variety of discrete neuronal subpopulations in various amygdalar nuclei via M2Rs, especially neurons that express SOM and NPY.

Postsynaptic targets of GABAergic basal forebrain projections to the basolateral amygdala.

Recent studies indicate that the basolateral amygdala, like the neocortex and hippocampus, receives GABAergic inputs from the basal forebrain in addition to the well-established cholinergic inputs. Since the neuronal targets of these inputs have yet to be determined, it is difficult to predict the functional significance of this innervation. The present study addressed this question in the rat by employing anterograde tract tracing combined with immunohistochemistry at the light and electron microscopic levels of analysis. Amygdalopetal axons from the basal forebrain mainly targeted the basolateral nucleus (BL) of the amygdala. The morphology of these axons was heterogeneous and included GABAergic axons that contained vesicular GABA transporter protein (VGAT). These axons, designated type 1, exhibited distinctive large axonal varicosities that were typically clustered along the length of the axon. Type 1 axons formed multiple contacts with the cell bodies and dendrites of parvalbumin-containing (PV+) interneurons, but relatively few contacts with calretinin-containing and somatostatin-containing interneurons. At the ultrastructural level of analysis, the large terminals of type 1 axons exhibited numerous mitochondria and were densely packed with synaptic vesicles. Individual terminals formed broad symmetrical synapses with BL PV+ interneurons, and often formed additional symmetrical synapses with BL pyramidal cells. Some solitary type 1 terminals formed symmetrical synapses solely with BL pyramidal cells. These results suggest that GABAergic neurons of the basal forebrain provide indirect disinhibition, as well as direct inhibition, of BL pyramidal neurons. The possible involvement of these circuits in rhythmic oscillations related to emotional learning, attention, and arousal is discussed.

Parvalbumin-immunoreactive neurons and GABAergic neurons of the basal forebrain project to the rat basolateral amygdala.

The basal forebrain (BF) contains a diffuse array of cholinergic and non-cholinergic neurons that project to the cerebral cortex and basolateral nuclear complex of the amygdala (BLC). Previous studies have shown that the GABAergic subpopulation of non-cholinergic corticopetal BF neurons selectively innervates cortical interneurons. Although several investigations in both rodents and primates have indicated that some BF neurons projecting to the BLC are non-cholinergic, there have been no studies that have attempted to identify the neurochemical phenotype(s) of these neurons. The present study combined Fluorogold retrograde tract tracing with immunohistochemistry for two markers of BF GABAergic neurons, parvalbumin (PV) or glutamic acid decarboxylase (GAD), to determine if a subpopulation of BF GABAergic cells projects to the BLC. Injections of Fluorogold confined to the rat BLC, and centered in the basolateral nucleus, produced extensive retrograde labeling in the ventral pallidum and substantia innominata regions of the BF. Although the great majority of retrogradely labeled neurons were not double-labeled, about 10% of these neurons, located mainly along the ventral aspects of the fundus striati and globus pallidus, exhibited immunoreactivity for PV or GAD. The results of this investigation contradict the long-held belief that there is no extra-amygdalar source of GABAergic inputs to the BLC, and indicate that the cortex-like BLC, in addition to the cortex proper, receives inhibitory inputs from the basal forebrain.

Immunohistochemical characterization of parvalbumin-containing interneurons in the monkey basolateral amygdala.

Interneurons expressing the calcium-binding protein parvalbumin (PV) are a critical component of the inhibitory circuitry of the basolateral nuclear complex (BLC) of the mammalian amygdala. These neurons form interneuronal networks interconnected by chemical and electrical synapses, and provide a strong perisomatic inhibition of local pyramidal projection neurons. Immunohistochemical studies in rodents have shown that most parvalbumin-positive (PV+) cells are GABAergic interneurons that co-express the calcium-binding protein calbindin (CB), but exhibit no overlap with interneuronal subpopulations containing the calcium-binding protein calretinin (CR) or neuropeptides. Despite the importance of identifying interneuronal subpopulations for clarifying the major players in the inhibitory circuitry of the BLC, very little is known about these subpopulations in primates. Therefore, in the present investigation dual-labeling immunofluorescence histochemical techniques were used to characterize PV+ interneurons in the basal and lateral nuclei of the monkey amygdala. These studies revealed that 90-94% of PV+ neurons were GABA+, depending on the nucleus, and that these neurons constituted 29-38% of the total GABAergic population. CB+ and CR+ interneurons constituted 31-46% and 23-27%, respectively, of GABAergic neurons. Approximately one quarter of PV+ neurons contained CB, and these cells constituted one third of the CB+ interneuronal population. There was no colocalization of PV with the neuropeptides somatostatin or cholecystokinin, and virtually no colocalization with CR. These data indicate that the neurochemical characteristics of the PV+ interneuronal subpopulation in the monkey BLC are fairly similar to those seen in the rat, but there is far less colocalization of PV and CB in the monkey. These findings suggest that PV+ neurons are a discrete interneuronal subpopulation in the monkey BLC and undoubtedly play a unique functional role in the inhibitory circuitry of this brain region.

Dopaminergic innervation of interneurons in the rat basolateral amygdala.

The basolateral nuclear complex of the amygdala (BLC) receives a dense dopaminergic innervation that plays a critical role in the formation of emotional memory. Dopamine has been shown to influence the activity of BLC GABAergic interneurons, which differentially control the activity of pyramidal cells. However, little is known about how dopaminergic inputs interface with different interneuronal subpopulations in this region. To address this question, dual-labeling immunohistochemical techniques were used at the light and electron microscopic levels to examine inputs from tyrosine hydroxylase-immunoreactive (TH+) dopaminergic terminals to two different interneuronal populations in the rat basolateral nucleus labeled using antibodies to parvalbumin (PV) or calretinin (CR). The basolateral nucleus exhibited a dense innervation by TH+ axons. Partial serial section reconstruction of TH+ terminals found that at least 43-50% of these terminals formed synaptic junctions in the basolateral nucleus. All of the synapses examined were symmetrical. In both TH/PV and TH/CR preparations the main targets of TH+ terminals were spines and distal dendrites of unlabeled cells. In sections dual-labeled for TH/PV 59% of the contacts of TH+ terminals with PV+ neurons were synapses, whereas in sections dual-labeled for TH/CR only 13% of the contacts of TH+ terminals with CR+ cells were synapses. In separate preparations examined in complete serial sections for TH+ basket-like innervation of PV+ perikarya, most (76.2%) of TH+ terminal contacts with PV+ perikarya were synapses. These findings suggest that PV+ interneurons, but not CR+ interneurons, are prominent synaptic targets of dopaminergic terminals in the BLC.

Neuronal localization of 5-HT type 2A receptor immunoreactivity in the rat basolateral amygdala.

Although it is well established that there are alterations in type 2A 5-HT receptors (5-HT2ARs) in the basolateral nuclear complex of the amygdala (BLC) in several neuropsychiatric disorders, very little is known about the neuronal localization of these receptors in this brain region. Single-labeling and dual-labeling immunohistochemical techniques were utilized in the rat to address this question. Three different 5-HT2AR antibodies were used, each producing distinct but overlapping patterns of immunostaining. Two of three 5-HT2AR antibodies mainly stained pyramidal projection neurons in the BLC. The third antibody only stained pyramidal cells in the dorsolateral subdivision of the lateral amygdalar nucleus. With one of the antibodies, the most intensely stained neurons were a population of large nonpyramidal neurons whose morphology and distribution closely resembled those shown in previous studies to project to the mediodorsal thalamic nucleus (MD). This was confirmed in the present study using a technique that combined 5-HT2AR immunohistochemistry with fluorogold retrograde tract-tracing. Two of three 5-HT2AR antibodies stained large numbers of parvalbumin-containing interneurons in the BLC. One of these two antibodies also stained a subpopulation of somatostatin-containing neurons. None of the 5-HT2AR antibodies stained significant numbers of the other two main interneuronal subpopulations, the large cholecystokinin-positive neurons or the small interneurons that exhibit extensive colocalization of calretinin and cholecystokinin. Since each of the three antibodies was raised against a distinct immunizing antigen, they may recognize different conformations of 5-HT2AR in different neuronal domains. The expression of 5-HT2ARs in pyramidal cells and parvalbumin-positive interneurons in the BLC is consistent with the results of previous electrophysiological studies, and suggests that 5-HT may produce excitation of several neuronal populations in the BLC via 5-HT2ARs.

A novel subpopulation of 5-HT type 3A receptor subunit immunoreactive interneurons in the rat basolateral amygdala.

The amygdalar basolateral nuclear complex (BLC) has very high levels of the 5-HT type 3 receptor (5-HT(3)R). Previous studies have reported that 5-HT(3)R protein in the BLC is expressed in interneurons and that 5-HT(3)R mRNA is coexpressed with GABA and certain neuropeptides or calcium-binding proteins in these cells. However, there have been no detailed descriptions of the distribution of 5-HT(3)R+ neurons in the rat amygdala, and no quantitative studies of overlap of neurons expressing 5-HT(3)R protein with distinct interneuronal subpopulations in the BLC. The present investigation employed dual-labeling immunohistochemistry using antibodies to the 5-HT-3A receptor subunit (5-HT(3A)R) and specific interneuronal markers to address these questions. These studies revealed that there was a moderate density of nonpyramidal 5-HT(3A)R+ neurons in the BLC at all levels of the amygdala. In addition, immunostained cells were also seen in anterior portions of the cortical and medial nuclei. Although virtually all 5-HT(3A)R+ neurons in the BLC were GABA+, very few expressed neuropeptide or calcium-binding protein markers for individual subpopulations. The main interneuronal marker expressed by 5-HT(3A)R+ neurons was cholecystokinin (CCK), but only 8-16% of 5-HT(3)R+ neurons in the BLC, depending on the nucleus, were CCK+. Most of these CCK+/5-HT(3A)R+ double-labeled neurons appeared to belong to the subpopulation of large type L CCK+ interneurons. Very few 5-HT(3A)R+ neurons expressed calretinin, vasoactive intestinal peptide, or parvalbumin, and none expressed somatostatin or calbindin. Thus, the great majority of neurons expressing 5-HT(3A)R protein appear to constitute a previously unrecognized subpopulation of GABAergic interneurons in the BLC.

Differential expression of Kv3.1b and Kv3.2 potassium channel subunits in interneurons of the basolateral amygdala.

The expression of Kv3.1 and Kv3.2 voltage-gated potassium channel subunits appears to be critical for high-frequency firing of many neuronal populations. In the cortex these subunits are mainly associated with fast-firing GABAergic interneurons containing parvalbumin or somatostatin. Since the basolateral nuclear complex of the amygdala contains similar interneurons, it is of interest to determine if these potassium channel subunits are expressed in these same interneuronal subpopulations. To investigate this issue, peroxidase and dual-labeling fluorescence immunohistochemistry combined with confocal laser scanning microscopy was used to determine which interneuronal subpopulations in the basolateral nuclear complex of the rat amygdala express Kv3.1b and Kv3.2 subunits. Antibodies to parvalbumin, somatostatin, calretinin, and cholecystokinin were used to label separate subsets of basolateral amygdalar interneurons. Examination of immunoperoxidase preparations suggested that the expression of both channels was restricted to nonpyramidal interneurons in the basolateral amygdala. Somata and proximal dendrites were intensely-stained, and axon terminals arising from presumptive basket cells and chandelier cells were lightly stained. Immunofluorescence observations revealed that parvalbumin+ neurons were the main interneuronal subpopulation expressing the Kv3.1b potassium channel subunit in the basolateral amygdala. More than 92-96% of parvalbumin+ neurons were Kv3.1b+, depending on the nucleus. These parvalbumin+/Kv3.1b+ double-labeled cells constituted 90-99% of all Kv3.1b+ neurons. Parvalbumin+ neurons were also the main interneuronal subpopulation expressing the Kv3.2 potassium channel subunit. More than 67-78% of parvalbumin+ neurons were Kv3.2+, depending on the nucleus. However, these parvalbumin+/Kv3.2+ double-labeled cells constituted only 71-81% of all Kv3.2+ neurons. Most of the remaining neurons with significant levels of the Kv3.2 subunit were somatostatin+ interneurons. These Kv3.2-containing somatostatin+ interneurons constituted 27-50% of the somatostatin+ population, depending on the nucleus in question. These data suggest that both fast-firing and burst-firing parvalbumin+ interneurons in the basolateral amygdala express the Kv3.1b subunit. The significance of Kv3.2 expression in some parvalbumin+ and somatostatin+ interneurons remains to be determined.

Localization of the CB1 type cannabinoid receptor in the rat basolateral amygdala: high concentrations in a subpopulation of cholecystokinin-containing interneurons.

The neuronal localization of the CB1 cannabinoid receptor in the rat basolateral amygdala was studied using peroxidase and fluorescence immunohistochemical techniques. All nuclei of the basolateral amygdala contained a large number of lightly stained pyramidal neurons and a small number of more intensely stained non-pyramidal neurons. Most of the latter cells had medium-sized to large multipolar somata and three to four aspiny dendrites, but some exhibited smaller oval somata. The axon initial segments of some of these non-pyramidal neurons exhibited large swollen varicosities in colchicine-injected animals, suggesting that much of the CB1 receptor protein is transported down the axons of these cells. Double-labeling studies using immunofluorescence histochemistry combined with confocal laser scanning microscopy revealed that the great majority of non-pyramidal neurons with CB1 receptor immunoreactivity belonged to a cholecystokinin-containing subpopulation. Whereas none of the other subpopulations of non-pyramidal neurons (exhibiting immunoreactivity for calretinin, parvalbumin, or somatostatin) expressed high levels of CB1 receptor immunoreactivity, a small percentage of these cells exhibited low levels of immunoreactivity. The results indicate that cannabinoids may modulate the activity of pyramidal projection neurons as well as a subpopulation of cholecystokinin-containing non-pyramidal neurons in the basolateral amygdala. Previous studies indicate that most of the latter are inhibitory interneurons that utilize GABA as a neurotransmitter. The intense staining of the cholecystokinin-containing interneurons and the evidence that large amounts of CB1 receptor protein are transported down the axons of these cells suggests that, as in the hippocampus, cannabinoids may inhibit the release of GABA from the axon terminals of these neurons.

Colocalization of calcium-binding proteins and GABA in neurons of the rat basolateral amygdala.

The basolateral amygdala contains subpopulations of non-pyramidal neurons that express the calcium-binding proteins parvalbumin, calbindin-D28k (calbindin) or calretinin. Although little is known about the exact functions of these proteins, they have provided useful markers of specific neuronal subpopulations in studies of the neuronal circuitry of the cerebral cortex and other brain regions. The purpose of the present study was to investigate whether basolateral amygdalar non-pyramidal neurons containing parvalbumin, calbindin, or calretinin exhibit immunoreactivity for GABA, and to determine if calretinin is colocalized with parvalbumin or calbindin in the rat basolateral amygdala. Pyramidal neurons were distinguished from non-pyramidal neurons on the basis of staining intensity. Using immunofluorescence confocal laser scanning microscopy, as well as the 'mirror technique' on immunoperoxidase-stained sections, it was found that there was virtually no colocalization of calretinin with parvalbumin or calbindin, but that the great majority of basolateral amygdalar non-pyramidal neurons containing parvalbumin, calbindin, or calretinin exhibited GABA immunoreactivity. Calbindin-positive neurons constituted almost 60% of the GABA-containing population in both subdivisions of the basolateral nucleus and more than 40% of the GABA-containing population in the lateral nucleus. Parvalbumin-positive neurons constituted 19-43% of GABA-immunoreactive neurons in the basolateral amygdala, depending on the nucleus. Calretinin-positive non-pyramidal neurons constituted about 20% of the GABA-positive neuronal population in each nucleus of the basolateral amygdala. These findings indicate that non-pyramidal neurons containing parvalbumin, calbindin, or calretinin comprise the majority of GABA-containing neurons in the basolateral amygdala, and that the calretinin subpopulation is distinct from non-pyramidal subpopulations containing parvalbumin and calbindin. These separate neuronal populations may play unique roles in the inhibitory circuitry of the amygdala.

Projections of the lateral entorhinal cortex to the amygdala: a Phaseolus vulgaris leucoagglutinin study in the rat.

In addition to providing a gateway to the hippocampus, the entorhinal cortex has significant projections to the amygdala. In the present investigation, the organization of the projections of the lateral entorhinal cortex to the amygdala was studied in the rat using the sensitive anterograde tracer Phaseolus vulgaris leucoagglutinin. Each of the three main subdivisions of the lateral entorhinal cortex provided a characteristic projection to the amygdala that mainly arose from the deep cortical layers. The projections from the dorsolateral and ventrolateral entorhinal areas were much stronger than those arising from the ventromedial entorhinal area. The primary targets of the dorsolateral and ventrolateral entorhinal areas were the basolateral amygdala, lateral capsular subdivision of the central nucleus and caudal portions of the cortical nuclear complex. The dorsolateral entorhinal area projects mainly to the lateral part of the basal nucleus, while the ventrolateral entorhinal area projects mainly to its medial part. A transitional region at the rostral pole of the ventrolateral entorhinal cortex has additional strong projections to the lateral subdivision of the central nucleus, medial amygdaloid nucleus and the intra-amygdaloid portion of the bed nucleus of the stria terminalis. The results of the present study indicate that the amygdala is one of the principal targets of the entorhinal cortex. The correspondence between the topography of entorhino-hippocampal connections and entorhino-amygdaloid connections suggests that the amygdaloid projection arising in each of the three main subdivisions of the entorhinal cortex conveys information processed in different septotemporal portions of the hippocampal formation. These entorhinal projections, which probably convey complex relational (including contextual) information to the amygdala, are in a position to produce different behavioral responses by activating different portions of the amygdaloid nuclear complex.

Immunohistochemical localization of the beta 2 and beta 3 subunits of the GABAA receptor in the basolateral amygdala of the rat and monkey.

The basolateral amygdala has a strong intrinsic inhibitory system mediated by GABAA receptors and is the main site of the anxiolytic actions of benzodiazepines. In an effort to identify the anatomical substrates for these transmitter and drug actions, immunohistochemical techniques were used to analyse the neuronal localization of the beta 2 and beta 3 receptor subunits of the GABAA-benzodiazepine receptor complex in the rat and monkey basolateral amygdala. The overall pattern of GABAA-benzodiazepine receptor immunoreactivity was very similar in both species. The density of the immunoreactivity in the neuropil varied in different nuclei of the basolateral amygdaloid complex. In both species the neuropil of the lateral nucleus exhibited the most robust staining. Immunoreactivity was also seen in neuronal perikarya and dendrites where it was localized to the cytoplasm and/or surface membrane. The cell type with the strongest immunoreactivity was a subpopulation of small non-pyramidal neurons that had numerous thin dendrites. Other larger non-pyramidal neurons were also stained. Pyramidal neurons in the rat and monkey basolateral amygdala exhibited light to moderate perikaryal staining that varied in different nuclei. The results of this study indicate that the pattern of GABAA-benzodiazepine receptor immunoreactivity in the neuropil of the rat and monkey basolateral amygdala closely resembled the distribution of benzodiazepine receptors localized in previous radioligand autoradiographic studies. The finding of intense immunoreactivity in subpopulations of non-pyramidal neurons suggests the existence of disinhibitory mechanisms which may be important for the activation of basolateral amygdaloid projection neurons.

Cortico-cortical and cortico-amygdaloid projections of the rat occipital cortex: a Phaseolus vulgaris leucoagglutinin study.

The efferent projections of the occipital cortex of the rat were investigated using the Phaseolus vulgaris leucoagglutinin anterograde tract tracing technique. Particular attention was focused on projections to the amygdala and amygdalopetal cortical areas. The primary visual cortex had projections to the medial and lateral portions of occipital area 2 and other cortical regions, but no projections to the amygdala or amygdalopetal cortical areas. The only occipital area that had direct projections to the amygdala was the most ventral portion of lateral occipital area 2, located just dorsal to temporal area 2. This occipitotemporal junction region, which received projections from secondary visual cortical areas but not from the primary visual cortex, had projections to the lateral nucleus, magnocellular basal nucleus, and lateral capsular subdivision of the central nucleus of the amygdala. Occipital area 2 had projections to seven amygdalopetal cortical regions: temporal area 2, temporal area 3, frontal area 2, ventrolateral orbitofrontal area, occipitotemporal junction region, lateral entorhinal area, and the perirhinal cortex. Projections to the perirhinal cortex targeted regions located adjacent to the parietal cortex and caudal temporal cortex, but not regions adjacent to the rostral temporal cortex. Other cortical regions receiving projections from medial and lateral portions of occipital area 2 included the presubiculum, retrosplenial areas, and caudal portions of the parietal cortical areas 1 and 2. The results of the present investigation, in conjunction with previous anatomical and neurobehavioral studies, support the concept that rodent cortical visual pathways, like those of primates, consist of a dorsal system involved with visuospatial functions and a ventral system involved with object recognition. As in primates, the ventral pathway projects to the temporal-perirhinal region in a cascading manner; only highly processed information from tertiary visual cortical areas reaches the amygdala. Unlike primates, however, cortical areas in the rat brain that receive highly processed visual information appear to be regions of multisensory convergence.

Projections of the medial and lateral prefrontal cortices to the amygdala: a Phaseolus vulgaris leucoagglutinin study in the rat.

The projections of different subfields of the medial and lateral prefrontal cortices to the amygdala were studied in the rat using the sensitive Phaseolus vulgaris leucoagglutinin anterograde tract tracing technique. Injections into the infralimbic cortex produced anterograde labeling in the lateral capsular subdivision of the central nucleus, superficial (corticomedial) amygdaloid nuclei, lateral and accessory basal nuclei, and the anterior amygdaloid area. Injections into the caudal portion of the infralimbic cortex produced additional labeling in the intermediate subdivision of the central nucleus. The prelimbic cortex had projections to the medial portion of the magnocellular basal nucleus and adjacent portions of the lateral nucleus and lateral capsular subdivision of the central nucleus. The medial precentral cortex had projections to the rostromedial part of the magnocellular basal nucleus and adjacent portions of the lateral capsular subdivision of the central nucleus. Injections into the lateral orbital and ventral agranular insular cortices produced labeled fibers in the rostral part of the superficial amygdala, lateral capsular subdivision of the central nucleus, and the lateral and accessory basal nuclei. The dorsal agranular insular area had projections to several different subdivisions of the central nucleus as well as to the rostrolateral magnocellular basal nucleus; the latter projections were complementary to those originating in the prelimbic area. The present study indicates that each portion of the prefrontal cortex has a distinctive projection to the amygdala. The ventral areas of the lateral and medial prefrontal cortices, which receive olfactory projections, are the only prefrontal cortical areas with projections to the olfactory-related superficial amygdaloid nuclei. The more dorsally situated prefrontal areas, the dorsal agranular insular area and prelimbic cortex, have complementary projections to the basal nucleus, suggesting that they modulate separate prefrontal cortico-striatal-pallid circuits. The specificity of prefrontal cortico-amygdaloid projections is indicative of their involvement in discrete functions.

Synaptology of prefrontal cortical projections to the basolateral amygdala: an electron microscopic study in the rat.

Prefrontal projections to the magnocellular basal amygdaloid nucleus (Bmg) of the rat were investigated using Phaseolus vulgaris leucoagglutinin (PHA-L) as an anterograde tracer. Electron microscopic examination revealed that most axon terminals in Bmg labeled by PHA-L injections into the prelimbic area contained round synaptic vesicles and made asymmetric synapses. The great majority of labeled terminals (93%) made synaptic contact with dendritic spines; a few contacts (7%) were seen with thin dendrites. These findings indicate that the main postsynaptic targets of PFC afferents to Bmg are spiny pyramidal neurons, the projection neurons of the basolateral amygdala. The morphology of the synapses suggests that they are excitatory.

Neuropeptide Y and somatostatin-like immunoreactivity in neurons of the monkey amygdala.

Neurons in the monkey amygdala exhibiting neuropeptide Y-like immunoreactivity and somatostatin-like immunoreactivity were identified using an avidin-biotin immunohistochemical technique. Differential co-existence of the two peptides was demonstrated using two-color immunoperoxidase and adjacent section methods. Numerous neuropeptide Y-positive neurons were observed in the basolateral and superficial amygdaloid nuclei. A moderate number of neuropeptide Y-positive neurons was seen in the medial subdivision of the central nucleus, but only a few neurons were observed in the lateral subdivision. Numerous somatostatin-positive neurons were stained in all major amygdaloid nuclei and always outnumbered neuropeptide Y-positive cells. All amygdaloid nuclei contained numerous peptide-positive fibers whose density varied depending on the nucleus. Approximately 90% of neuropeptide Y-positive neurons also exhibited somatostatin-like immunoreactivity. The percentage of somatostatin-positive neurons that exhibited neuropeptide-Y immunoreactivity varied in different nuclei. In the superficial amygdaloid nuclei, medial subdivision of the central nucleus and most portions of the basolateral nuclei the predominant cell type stained with both the neuropeptide Y and somatostatin antibodies was a spine-sparse non-pyramidal neuron. In the dorsal portion of the lateral nucleus, however, most peptide-positive neurons had spiny dendrites. Only the cell bodies and proximal dendrites of somatostatin-positive neurons in the lateral subdivision of the central nucleus were immunostained. This study demonstrates that specific cell populations in the primate amygdala contain neuropeptide Y, somatostatin or both peptides. Most peptide-positive neurons in the basolateral and superficial amygdaloid nuclei appear to be local circuit neurons that contribute to the dense plexus of peptide-positive axons in these regions. The finding of neurons with spiny dendrites in the dorsal part of the lateral nucleus suggests that these cells may be functionally different from peptide-positive neurons in other portions of the basolateral amygdala. The lateral subdivision of the central nucleus is distinguished from other amygdaloid nuclei by containing a large population of somatostatin-positive neurons that do not exhibit neuropeptide Y immunoreactivity.

Cholecystokinin corticostriatal pathway in the rat: evidence for bilateral origin from medial prefrontal cortical areas.

The anterograde tracer Phaseolus vulgaris-leucoagglutinin was used to examine the organization of the projections to the striatum from medial prefrontal and frontal cortical areas in the rat with reference to their relation to cholecystokinin-like immunoreactivity in the striatum. Medial prefrontal cortical areas projected bilaterally, with an ipsilateral predominance, to the striatum. Most of the positive fibres were found in medial and ventral areas of the caudate-putamen and in the nucleus accumbens. Labelled fibres formed distinct patch-like arrangements throughout the dorsomedial striatum, whereas more ventrally the fibres were densely packed and spread to lateral areas. Almost no fibres were found in the dorsolateral aspects of the caudate-putamen. Cholecystokinin-like immunoreactivity in the striatum was diffusely distributed in the medial aspects, in fine punctate elements as well as in patches of fibres. Overlapping of corticostriatal clusters of fibres, from medial prefrontal cortex, with cholecystokinin-immunoreactive patches was found at all rostrocaudal levels studied, but predominantly in rostral areas. The overlap was present both in the ipsilateral and the contralateral side. Often the cluster of corticostriatal fibres was completely and precisely overlaid by a cholecystokinin-immunoreactive patch. At more caudal planes the overlap was only partial and in some instances cholecystokinin-positive patches "avoided" zones of dense corticostriatal fibre terminations. Frontal cortex injections of tracer gave rise to a network of fibres in the lateral aspects of the striatum, sparing the medial areas. No overlap with cholecystokinin-immunoreactive patches was found in these cases. These results suggest that a large number of cholecystokinin-containing striatal fibres originate in medial prefrontal cortical areas.

A sexually dimorphic population of CRF neurons in the medial preoptic area.

The neuropeptide corticotropin-releasing factor (CRF) is thought to mediate the induction of a constellation of behavioral, endocrine, and autonomic responses which are important for an animal's adaptation to stressful events. We have found that the anteroventral periventricular preoptic nucleus (AVPv) and medial preoptic nucleus (MPN) of colchicine-injected female rats contained numerous intensely stained CRF-immunoreactive neurons. The AVPv/MPN in males contained very few CRF-immunoreactive neurons per section, even in colchicine-injected animals. This sexually dimorphic population of CRF-immunoreactive neurons in the AVPv may play some role in the sex-related differences in hormonal responses to stress and/or in the control of female reproductive events.