Rat Amygdaloid Complex Research Articles

Paleoamygdala: Morphogenesis of the Posterior Cortical Nucleus of the Rat Amygdaloid Complex of the Brain during the Early Juvenile Period

Sex-related differences and the dynamic of formation of the posterior cortical nucleus of the rat amygdaloid complex were revealed in the early juvenile period by planimetric characteristics, numbers of neurons and glial cells, and glial and apoptotic indexes reflecting morphological restructuring on postnatal days 21, 24, 28, and 31.

Spike distributions of a population based hierarchical network of the rat amygdaloid complex

Background The amygdaloid nuclear complex (ANC) is a conglomerate of gray matter lying in the depth of the anteromedial temporal lobe ventral to the lentiform nucleus. ANC is critical for producing appropriate emotional and behavioral responses to biologically relevant sensory stimuli. It constitutes an essential link between sensory and limbic areas of the cerebral cortex and subcortical brain regions, such as the hypothalamus, brainstem, and striatum, that are responsible for eliciting emotional and motivational responses. The connectome of the rat ANC is modeled in neuroVIISAS (a platform for mapping, visualization and analysis of connectivity data from tracing studies) that is used to generate realistic large scale networks of all regions of the central nervous system. The models are transferred by a PyNEST interface to the simulation engine NEST and simulation results can be analyzed and visualized by neuroVIISAS.

Distribution of 5-HT2A receptor immunoreactivity in the rat amygdaloid complex and colocalization with γ-aminobutyric acid

The 5-HT2A receptor (5-HT2Ar) is located in a variety of excitatory and inhibitory neurons in many regions of the central nervous system and is a major target for atypical antipsychotic drugs. In the present study, an immunoperoxidase experiment was used to investigate the distribution of 5-HT2Ar immunoreactivity in the rat amygdaloid complex. In the basolateral amygdala, the colocalization of 5-HT2Ar with inhibitory transmitter γ-aminobutyric acid (GABA) was studied using double-immunofluorescence confocal microscopy. The staining pattern obtained was colchicine-sensitive. In fact, pretreatment with colchicine increased the number of 5-HT2Ar-immunoreactive somata. Accordingly, with the exception of the intercalated nuclei, the amygdaloid complex of colchicine-injected rats exhibited a high density of 5-HT2Ar-IR somata. Morphological analyses indicated that 5-HT2Ar was located on both excitatory and inhibitory neurons in the rat amygdaloid complex. In addition, double-immunofluorescence observations revealed that the great majority of GABA-immunoreactive neurons in the basolateral amygdala exhibited 5-HT2Ar immunoreactivity (66.3%–70.6% depending on the nucleus). These data help to clarify the complex role of the 5-HT2Ar in the amygdaloid complex suggesting that this receptor can regulate amygdaloid activity by acting on different neuronal populations.

Enkephalin co‐expression with classic neurotransmitters in the amygdaloid complex of the rat

This study aimed at characterizing the neurotransmitter phenotype of enkephalin neurons in the rat amygdaloid complex. We first established the detailed distribution of vesicular glutamate transporters 1 and 2 (VGLUT1 and -2) and glutamate decarboxylase 65 (GAD65) in the amygdala by using in situ hybridization. In the amygdaloid complex, GAD65 is strongly expressed in striatal-like divisions, namely, the anterior amygdaloid area, the central nucleus (CEA), the intercalated nuclei, and the dorsal part of the medial nucleus (MEA). VGLUT1 and -2 expression is mostly segregated to specific divisions of the amygdale, with VGLUT2 being expressed only in the MEA, the anterior cortical nucleus (COAa), and the anterior basomedial nucleus (BMAa), whereas VGLUT1 is expressed in all other divisions of the amygdala. Second, we assessed the co-expression of preproenkephalin (ppENK) with GAD65, VGLUT1, or VGLUT2 by using double fluorescent in situ hybridization. We found that ppENK mRNA co-localized exclusively with GAD65 in all striatal-like structures of the amygdaloid complex with the exception of the MEA, where ENK also co-localized with VGLUT2 mRNA. This co-localization is most apparent in the posteroventral part of the MEA, where 70% of ENKergic cells expressed VGLUT2. In addition, ppENK also co-localized with VGLUT1 because more than 95% of ENK cells in the basolateral amygdala expressed VGLUT1. In contrast, less than 25% of ENKergic cells expressed VGLUT1 in the lateral nucleus of the amygdale, with the majority of ENK cells expressing GAD65 mRNA in this nucleus. These results have broad implications for understanding the functional roles of enkephalinergic neurotransmission in the amygdaloid complex.

Morphology, size and distribution of corticotropin releasing factor (CRF) immunoreactive neurons in the central nucleus of the rat amygdaloid complex

The amygdaloid complex (AC) is a heterogenous group of cortical and nuclear structures. As an important component of the limbic system contains numerous neurotransmitters including neuropeptides. In this study we have investigated the types, distribution, and morphometric characteristics of corticotropin releasing factor (CRF) immunoreactive neurons in the central nucleus (Ce) of the AC. We have also identified CRF- immunoreactive fibers in this nucleus. This research was performed on 5 adult rats kept in standard conditions. Animals were perfused 48 hours after application of colhicin. The removed brains were postfixed and cut into free floating sections. The sections were treated with antibodies against the rabbit CRF and ABC immunohistochemstry was applied. The neurons were measured and drawn with a camera lucida. We found uniform distribution of CRF immunoreactive neurons in the Ce of AC of rat. According to morphological type dominant were triangular (50%), while bipolar (25%) and multipolar (25%) types were also present. The most of the CRF immunoreactive neurons had 2-5 primary dendrites. These findings will contribute to understanding the role of CRF neurons in rat Ce during stress and in other behavioral functions.

Distribution of CB1 cannabinoid receptors in the rat amygdaloid complex

The amygdaloid complex (AK) has a very important role in the modulation of endocrine and visceral functions, in complex behavioral mechanisms such as defense, feeding, aggression, affects, reproduction, memory and learning. The aim of this study was to determine the precise distribution of cannabinoid CB1 receptors in the rat AK, using the immunohistochemical (ABC) method. According to our results, CB1-immunoreactivity in the rat AK was highest in the medial nucleus. Slightly lower immunoreactivity was found in the basolateral nucleus. Moderate density of CB1 receptors occurred in the central, basomedial, lateral and posterior nuclei of the AK. CB1-immunoreactivity in all of these nuclei was present in the form of discrete spot-like precipitates of unequal size and appearance. These precipitates exhibited three different patterns: 1. elongated columns or lines, 2. complete or incomplete rings and 3. in a small number of AK regions CB1-immunoreactivity was separately dispersed in the form of single spot-like precipitates between more complex columns and rings of precipitates. Considering the functional importance of amygdala and the distribution of CB1 receptors in the AK we could conclude that our findings suggest a role for cannabinoids in modulating responses of the AK to stress and fear as well as to pain.

Distribution of parvalbumin, calretinin, and calbindin-D(28k) immunoreactivity in the rat amygdaloid complex and colocalization with gamma-aminobutyric acid.

To understand the organization of inhibitory circuitries in the rat amygdala, the distribution of parvalbumin, calretinin, and calbindin immunoreactivity was investigated in the rat amygdaloid complex. Colocalization of various calcium-binding proteins with the inhibitory transmitter gamma-aminobutyric acid (GABA) was studied by using the mirror technique. Parvalbumin-immunoreactive (-ir) elements were located mostly in the deep amygdaloid nuclei, whereas the calretinin-ir and calbindin-ir staining were most intense in the cortical nuclei as well as in the central nucleus and the amygdalohippocampal area. Second, the distribution of immunopositive neurons largely parallelled the distribution of terminal and neuropil labeling. Third, immunostained neurons could be divided into four major morphologic types (types 1-4) based on the characteristics of the somata and the dendritic trees. The fourth lightly stained neuronal type that had a pyramidal GABA-negative soma was observed only in calretinin and calbindin preparations. Fourth, parvalbumin-ir terminals formed basket-like plexus and cartridges, which suggests that parvalbumin labels GABAergic inhibitory basket cells and axo-axonic chandelier cells, respectively. Colocalization studies indicated that 521 of 553 (94%) of parvalbumin-ir, 419 of 557 (75%) of calbindin-ir, and 158 of 657 (24%) of calretinin-ir neurons were GABA-positive in the deep amygdaloid nuclei. A high density of large GABA-negative calbindin-ir neurons was observed caudally in the medial division of the lateral nucleus and GABA-negative calretinin-ir neurons were observed in the magnocellular division of the accessory basal nucleus as well as in the intermediate and parvicellular divisions of the basal nucleus. These data suggest that in various amygdaloid areas, neuronal excitability is controlled by GABAergic neurons that contain different calcium-binding proteins. The appearance of basket-like plexus and cartridges in the parvalbumin preparations, but not in calretinin preparations, suggests that like in the hippocampus, the distribution of inhibitory terminals in the dendritic and perisomatic regions of postsynaptic neurons in the rat amygdala is organized in a topographic manner.

Status epilepticus-induced neuronal damage in the rat amygdaloid complex: distribution, time-course and mechanisms

The present study was designed to elucidate the distribution, time-course and mechanism(s) of status epilepticus-induced neuronal damage in the rat amygdaloid complex. Status epilepticus was induced with kainate (9 mg/kg, i.p.), and the behavioral and electrographic seizure activity of each rat was monitored via cortical electrodes attached to a continuous video electrocorticogram system. Rats were subsequently perfused 1, 2, 4, 8, 16, 24 or 48 h after kainate injection. The first signs of amygdaloid damage were seen in rats perfused 4 h after kainate injection, though the severity and temporal appearance of damage varied substantially between the different amygdaloid nuclei and their subdivisions. Second, terminal transferase dUTP nick-end labeling (TUNEL)-positive nuclei and laddering of DNA in gel electrophoresis appeared in the amygdala 8 and 16 h after kainate, respectively. The distribution and density of TUNEL-positive nuclei in the different amygdaloid nuclei correlated with the distribution of neuronal damage in Thionin- and silver-stained sections. Third, the immunoreactivity of Bax protein, a promoter of apoptotic neuronal death, increased in the vulnerable medial division of the lateral nucleus prior to the appearance of argyrophilic neurons and TUNEL-positive nuclei. Fourth, the severity of neuronal damage progressed in some, but not all, amygdaloid regions throughout the 48-h follow-up, even though the occurrence of high-amplitude and frequency discharges, which are typically associated with behavioral seizure activity, extinguished after 7 h. These data show that status epilepticus-induced neuronal damage in the amygdala is a dynamic region-specific process, the severity of which depends on the duration of seizure activity. At least one mechanism underlying the damage involves apoptosis, which continues long after the behavioral and electrographic seizures have subsided.

Intrinsic connections of the rat amygdaloid complex: projections originating in the central nucleus.

Inputs from the amygdaloid and extraamygdaloid areas terminate in various divisions of the central nucleus. To elucidate the interconnections between the different regions of the central nucleus and its connectivity with the other amygdaloid areas, we injected the anterograde tracer, Phaseolus vulgaris-leucoagglutinin (PHA-L) into the capsular, lateral, intermediate, and medial divisions of the central nucleus in rat. There were a number of labeled terminals near the injection site within each division. The intrinsic connections between the various divisions of the central nucleus were organized topographically and originated primarily in the lateral division, which projected to the capsular and medial divisions. Most of the connections were unidirectional, except in the capsular division, which received a light reciprocal projection from its efferent target, the medial division. The intermediate division did not project to any of the other divisions of the central nucleus. Extrinsic projections from the central nucleus to the other amygdaloid nuclei were meager. Light projections were observed in the parvicellular division of the basal nucleus, the anterior cortical nucleus, the amygdalohippocampal area, and the anterior amygdaloid area. No projections to the contralateral amygdala were found. These data show that the central nucleus has a dense network of topographically organized intradivisional and interdivisional connections that may integrate the intraamygdaloid and extraamygdaloid information entering the different regions of the central nucleus. The sparse reciprocal connections to the other amygdaloid nuclei suggest that the central nucleus does not regulate the other amygdaloid regions but, rather, executes the responses evoked by the other amygdaloid nuclei that innervate the central nucleus.

Differential distribution and regulation of estrogen receptor-α and -β mRNA within the female rat brain

In the present study, estrogen receptor (ER)α and ERβ genes were found to be differentially expressed in discrete subregions of the rat amygdaloid complex. The amygdala nuclei showing predominant ERα mRNA expression included the posterolateral cortical nucleus, amygdala hippocampal area, and lateral dorsolateral nucleus, whereas the amygdala areas with predominant ERβ mRNA expression were the medial anterodorsal and central nuclei. Both ERα and ERβ mRNAs were highly expressed in the medial posterodorsal nucleus. In addition to the discrete anatomical expression patterns, there appeared to be a differential regulation by estradiol of the ERα and ERβ mRNAs. Two weeks of estradiol (170 μg total) treatment decreased ERα mRNA expression levels in the arcuate, ventromedial hypothalamus, and posterolateral cortical amygdala nucleus, but increased ERβ mRNA in the arcuate. In the medial amygdala nuclei, only ERβ mRNA levels were altered (reduced) by estradiol treatment. These results suggest that estrogen can modulate behaviors and functions mediated by the amygdala and hypothalamus via differentially regulated ER subtypes.

Ultrastructural features of tyrosine-hydroxylase-immunoreactive afferents and their targets in the rat amygdala.

Interrelations of tyrosine-hydroxylase-immunoreactive afferent fibres with neuronal elements were studied in central, basal and intercalated nuclei of the rat amygdaloid complex. Comparison with dopamine-beta-hydroxylase-immunoreacted and phenylethanolamine-N-methyltransferase-immunoreacted parallel sections indicated that the tyrosine-hydroxylase immunoreaction labelled preferentially dopaminergic axons. At the electron-microscopic level, the majority of tyrosine-hydroxylase-immunoreactive axons possessed small boutons containing small clear vesicles and contacting dendrites, spines or somata of amygdala neurons, forming mostly symmetric synapses. They were often directly apposed to or in the vicinity of unlabelled terminals synapsing on the same structure. Synaptic density was highest in the central lateral part of the central nucleus. In the central and basal nuclei labelled axons synapsed preferentially on small dendrites and dendritic spines, and on somata of a few neurons. A detailed study of the neuronal ultrastructure showed that innervated somata possessed the differential characteristics displayed by the predominant neuron types in the medial and central lateral central nucleus and resembled the typical projection neurons in the basal nuclei. In the paracapsular intercalated cell groups the majority of neurons possessed intense perisomatic innervation by immunoreactive terminals. The results suggest that tyrosine-hydroxylase-immunoreactive, predominantly dopaminergic amygdaloid afferent fibres preferentially modulate the effect of extrinsic inputs into neurons of the central and basal nuclei, while a nonselective regulation is exerted upon the output of paracapsular intercalated neurons. It is suggested that this innervation pattern may be important for the coordinated integration of extrinsic and intraamygdaloid connections and thus for balanced output of the structure.

Decrease in somatostatin-immunoreactive neurons in the rat amygdaloid complex in a kindling model of temporal lobe epilepsy.

In human temporal lobe epilepsy, seizures can begin in the hippocampus, amygdala, or surrounding cortical areas. Histologically, the seizure-induced selective neuronal damage and synaptic reorganization are best documented in the hippocampus. Little information is available about the damage in the other temporal lobe structures or whether the distribution of damage depends on the location of the primary seizure focus. We used an amygdala-kindling model of temporal lobe epilepsy to study whether seizures of amygdaloid origin cause damage to the amygdala and hippocampus. All rats experienced five class 5 generalized seizures. Neuronal damage was assessed by counting the density of GABA-immunoreactive (GABA-ir) and somatostatin-immunoreactive (SOM-ir) neurons in the amygdala and hilus of the dentate gyrus six months after the last seizure. We found that the density of GABA-ir neurons did not differ from that in controls in the contralateral amygdala. The density of SOM-ir neurons was, however, decreased in the lateral (69% of neurons remaining, P < 0.01), basal (67% remaining, P < 0.05), and accessory basal (68% remaining, P < 0.05) nuclei. In the hilus, the densities of GABA-ir and SOM-ir neurons were similar to that in controls. According to our data, a few seizures of amygdaloid origin may cause more severe damage to SOM-ir neurons in the amygdala than in the hilus. Such decrease in SOM-ir neurons which form one subpopulation of GABAergic inhibitory interneurons may increase the local excitability in the amygdala and, therefore, contribute to epileptogenesis.

Status epilepticus causes selective regional damage and loss of GABAergic neurons in the rat amygdaloid complex.

In human epilepsy, the amygdala is often a primary focus for seizures. To analyse the status epilepticus-induced alterations in the amygdaloid circuitries which may later underlie epileptogenesis, we studied the amygdaloid damage in kainic acid and perforant pathway stimulation models of status epilepticus in the rat. We also studied the damage to inhibitory GABAergic neurons. In both models, the medial division of the lateral nucleus, the parvicellular division of the basal nucleus and portions of the anterior cortical and medical nuclei were damaged. In the kainate model, where the seizure activity was more severe, the accessory basal nucleus, amygdalohippocampal area, posterior cortical nucleus and periamygdaloid cortex were also damaged. Two weeks after kainate-induced seizures, 56% of the GABA-immunoreactive neurons remained in the lateral nucleus (P < 0.05) and 25% in the basal nucleus (P < 0.01). Further analysis showed that one subpopulation of damaged GABAergic neurons was immunoreactive for somatostatin (48% remaining in the lateral nucleus, P < 0.01; 33% in the basal nucleus, P < 0.01). In the perforant pathway stimulation model, the damage to somatostatin neurons was milder. According to our data, the initial insult, such as status epilepticus, selectively damages amygdaloid nuclei. The loss of inhibition may underlie the spontaneous generation of seizures and epileptogenesis. On the other hand, many amygdaloid output nuclei (magnocellular and intermediate division of the basal nucleus, the central nucleus) remained relatively undamaged, providing pathways for seizures spread and generation of seizure-related behavioural manifestations such as motor convulsions and fear response.

Interamygdaloid projections of the basal and accessory basal nuclei of the rat amygdaloid complex

Previous studies suggest that the left and right amygdalae are interconnected in rodents. The origin and topography of these connections have, however, remained obscure. In the present study, we investigated the interamygdaloid projections originating in the different divisions of the basal and accessory basal nuclei of the rat amygdala by using the Phaseolus vulgaris leucoagglutinin anterograde tract-tracing technique. The basal nucleus gave rise to substantial interamygdaloid projections. However, the density of the projections depended on the location of Phaseolus vulgaris leucoagglutinin injection in the basal nucleus. The magnocellular and intermediate divisions projected heavily to the homonymous regions on the contralateral side, as well as to the nucleus of the lateral olfactory tract. The parvicellular division projected lightly to the homonymous region on the contralateral side, to the contralateral anterior amygdaloid area and to the medial division of the central nucleus. The contralateral projections originating in the accessory basal nucleus were light compared to those of the basal nucleus.These data indicate that interamygdaloid connections in the rat brain are extensive and topographically organized. Via these connections, one amygdala may rapidly activate the contralateral side. This may explain, for example, why the epileptic seizures in one amygdala spread contralaterally and cause the development of independent seizure activity in kindling and in clinical epilepsy.

Intrinsic connections of the rat amygdaloid complex: projections originating in the accessory basal nucleus.

The amygdaloid complex plays an important role in the detection of emotional stimuli, the generation of emotional responses, the formation of emotional memories, and perhaps other complex associational processes. These functions depend upon the flow of information through intricate and poorly understood circuitries within the amygdala. As part of an ongoing project aimed at further elucidating these circuits, we examined the intra-amygdaloid connections of the accessory basal nucleus in the rat. In addition, we examined connections of the anterior cortical nucleus and amygdalahippocampal area to determine whether portions of these nuclei should be included in the accessory basal nucleus (as some earlier studies suggest). Phaseolus vulgaris leucogglutinin was injected into different rostrocaudal levels of the accessory basal nucleus (n = 12) or into the anterior cortical nucleus (n = 3) or amygdalahippocampal area (n = 2). The major intra-amygdaloid projections from the accessory basal nucleus were directed to the medial and capsular divisions of the central nucleus, the medial division of the amygdalohippocampal area, the medial division of the lateral nucleus, the central division of the medial nucleus, and the posterior cortical nucleus. The projections originating in the anterior cortical nucleus and the lateral division of the amygdalohippocampal area differed from those originating in the accessory basal nucleus, which suggests that these areas are not part of the accessory basal nucleus. The present findings and our previous data suggest that each of the deep amygdaloid nuclei have different intra-amygdaloid connections. The pattern of these various connections suggests that information entering the amygdala from different sources can be integrated only in certain amygdaloid regions.

Intrinsic connections of the rat amygdaloid complex: projections originating in the basal nucleus.

The amygdaloid complex is involved in associational processes, such as the formation of emotional memories about sensory stimuli. However, the anatomical connections through which the different amygdaloid nuclei process incoming information and communicate with the other amygdaloid nuclei, is poorly understood. As part of an ongoing project aimed at elucidating the intrinsic connections of the rat amygdaloid complex, we injected the anterograde tracer PHA-L (Phaseolus vulgaris-leucoagglutinin) into different rostrocaudal levels of the basal nucleus of the amygdala in 21 rats and analyzed the distribution of labeled fibers and terminals throughout the amygdaloid complex. The connectional analysis, together with cytoarchitectonic observations, suggested that contrary to previous notions the basal nucleus in the rat has three divisions: magnocellular, intermediate, and parvicellular. The magnocellular division has heavy reciprocal connections with the lateral portion of the parvicellular division and the intermediate division projects weakly to the parvicellular division, whereas the projection from the medial portion of the parvicellular division to the intermediate division is heavy and the lateral and medial portions of the parvicellular division are only weakly interconnected, as are the magnocellular and intermediate divisions. The main intraamygdaloid targets of the basal nucleus projections are the nucleus of the lateral olfactory tract, the anterior amygdaloid area, the medial and capsular divisions of the central nucleus, the anterior cortical nucleus, and the amygdalohippocampal area. Our findings provide the most detailed understanding of the intra-amygdala connections of the basal nucleus to date and show that the connections within the basal nucleus and between the basal nucleus and other amygdaloid areas are more widespread and topographically organized than previously recognized.

Intrinsic connections of the rat amygdaloid complex: Projections originating in the lateral nucleus

The amygdaloid complex receives sensory information from a variety of sources. A widely held view is that the amygdaloid complex utilizes this information to orchestrate appropriate species-specific behaviors to ongoing experiences. Relatively little is known, however, about the circuitry through which information is processed within the amygdaloid complex. The lateral nucleus is the major recipient of extrinsic sensory information and is the origin of many intra-amygdaloid projections. In this study, we reinvestigated the organization of intra-amygdaloid projections originating from the lateral nucleus using the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L). The lateral nucleus has highly organized intranuclear connections. Dense projections interconnect rostral and caudal levels of the lateral and the medial divisions of the nucleus, and the lateral and medial divisions of the lateral nucleus are also interconnected. The major extranuclear projections of the lateral nucleus are (in descending order of magnitude) to the accessory basal nucleus, the basal nucleus, the periamygdaloid cortex, the dorsal portion of the central division of the medial nucleus, the posterior cortical nucleus, the capsular division of the central nucleus, and the lateral division of the amygdalohippocampal area. The pattern of extranuclear projections varied depending on the rostrocaudal or mediolateral location of the injection site within the lateral nucleus. These findings indicate that intra-amygdaloid projections originating in the lateral nucleus are both more widespread and more topographically organized than was previously appreciated.

Distribution of neurons of origin of zinc-containing projections in the amygdala of the rat

The present study describes the distribution of neurons of origin of zinc-containing pathways in the amygdaloid complex of the rat, using the selenium method for simultaneous retrograde labeling of all zinc-containing neurons. With this method, vesicular ionic zinc is precipitated intravitally with selenium compounds and transported retrogradely to the parent neurons, where it can be visualized by silver amplification. Neurons labeled retrogradely with silver-amplified precipitate were observed in all amygdaloid nuclei except for the lateral olfactory tract nucleus, the accessory olfactory tract nucleus and the central nucleus. Very few labeled cell bodies were seen in the anterior amygdaloid area and the medial nucleus. The amygdalo-hippocampal area and the amygdalo-piriform transition area both showed a substantial number of labeled somata throughout their rostrocaudal extent. In the anterior cortical nucleus, very few labeled cell bodies were found in the rostral pole, whereas they were abundant in the caudal quarter of the nucleus. In the posterolateral cortical nucleus, the number of labeled cell bodies increased gradually; there were none in the rostral pole, but most of the neurons in the caudal part were labeled. The posteromedial cortical nucleus contained a great number of labeled somata, but with some variation in the rostrocaudal extent of the nucleus. Considerable numbers of labeled neurons were observed throughout the lateral nucleus. In the basolateral nucleus, a small number of labeled cell bodies was present in the rostral half, but a gradual increase was observed in the caudal direction. Finally, in the basomedial nucleus, very few labeled cell bodies were present in the rostral two-thirds, whilst a considerable number was encountered in the caudal one-third. Possible functional implications of neuronal zinc are considered. The distribution of neurons of origin of zinc-containing projections has been compared with previously described intrinsic connections of the rat amygdala, and tracts that may possibly be zinc-containing are outlined and discussed. It is concluded that in all probability a substantial proportion of the intrinsic connectivity of the rat amygdaloid complex is zinc-containing.

Multiple high-threshold calcium currents in acutely isolated rat amygdaloid pyramidal cells.

1. High-threshold Ca2+ currents were studied in large pyramidal cells acutely dissociated from the rat amygdaloid complex. L-, N-, and P-type currents were present in about equal proportions in these cells and accounted for most of the whole-cell current.

Comparative single and double immunolabelling with antisera against catecholamine biosynthetic enzymes: criteria for the identification of dopaminergic, noradrenergic and adrenergic structures in selected rat brain areas

Immunodetection of catecholamine biosynthetic enzymes is frequently used for the visualization of central nervous catecholaminergic systems. Because of the method's limited specificity for the different catecholamines, interpretation of the results often presents difficulties. To determine criteria for the identification of dopaminergic, noradrenergic, and adrenergic afferents to the rat amygdaloid complex, comparative immunolabelling for tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH), and phenylethanolamine-N-methyl-transferase (PNMT) was carried out using single- and double-labelling for fluorescence, light- and electron microscopy. The observations were complemented by findings in brainstem and hypothalamic areas. The results indicated that TH-labelling detected preferentially dopaminergic afferents in the lateral central and intercalated amygdaloid nuclei. DBH-labelling detected noradrenergic axons in nuclei lacking PNMT-immunoreactive fibres, and PNMT was a marker for adrenergic axons in the entire complex. For nuclei with combined dense dopaminergic, noradrenergic and/or adrenergic innervation, morphological and immunolabelling characteristics were described which, to a certain extent, enabled identification of the different afferents in anti-TH or anti-DBH-preparations. Using a monoclonal TH-antiserum, noradrenergic and adrenergic axons displayed weaker immunoreactivity than dopaminergic ones, and possessed characteristic morphological features. TH-immunoreactivity in noradrenergic axons differed depending on their origin, and showed intra-axonal compartmentalization. The present study provides a basis for the use of the detection of biosynthetic enzymes in future investigations into the ultrastructure and connectivity of the catecholaminergic amygdala innervation.