Evidence for a GABAergic interface between cortical afferents and brainstem projection neurons in the rat central extended amygdala

The synaptic circuitry of the intrinsic GABAergic system of the central extended amygdala (CEA) in relation to efferent neurons and cortical afferents was examined in the present study. Neurons in the CEA projecting to the dorsal vagal complex and the parabrachial complex were identified by the retrograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Postembedding GABA-immunocytochemistry revealed that GABA-immunoreactive (GABA-IR) terminals formed largely symmetrical synaptic contacts with the perikarya and proximal dendritic processes of almost all WGA-HRP-labeled neurons in the CEA. To determine the relationship between cortical afferents and CEA GABAergic neurons, WGA-HRP was used to anterogradely label afferents from the insular cortex in combination with postembedding immunogold detection of GABA. Cortical afferents formed asymmetrical synaptic contacts predominantly on small dendrites and dendritic spines. Many of the dendrites postsynaptic to cortical terminals in the central nucleus were immunoreactive for GABA although only relatively few spines were GABA-IR. Combining pre-embedding GAD-immunocytochemistry with cortical lesions resulted in approximately 40% of degenerating terminals of insular cortical origin in the central nucleus in contact with small, GAD-IR dendrites and spines. The present results demonstrate that the neurons providing the major CEA outputs to the brainstem receive an extensive GABAergic innervation, strongly supporting our proposal that CEA efferent neurons are under strong tonic inhibition by intrinsic GABAergic neurons. Further, our finding that the major cortical input to the central nucleus preferentially innervates intrinsic GABAergic neurons suggests that these neurons in the CEA may serve as an interface between the principal inputs and outputs of this forebrain region. © Wiley-Liss, Inc.     

Intrinsic and extrinsic connections of the rat central extended amygdala: an in vivo electrophysiological study of the central amygdaloid nucleus

Anatomical studies have shown that the central amygdaloid nucleus (CeA) is reciprocally connected with the lateral bed nucleus of the stria terminalis (BSTL), both structures being major components of the central extended amygdala. The CeA also receives projections from the insular cortex (InsCx) and the paraventricular thalamic nucleus (PVT). Extracellular unit activity was recorded from neurons in the lateral CeA (CeL) in urethane anaesthetized rats and their responses were studied after electrical stimulation of the BSTL, InsCx and PVT. The spontaneous activity of CeL neurons was low (1.69 spikes/s) and 40% of recorded cells were silent. The iontophoretic application of the GABA_A antagonist, bicuculline, increased the firing rate of 20% of neurons. The BSTL stimulation induced an antidromic response in 33% of the tested cells. Orthodromic responses were obtained from 83% (BSTL stimulation), 70% (InsCx stimulation) and 85% (PVT stimulation) of tested cells, some of which responded to both BSTL and InsCx or PVT stimulations. Orthodromic responses mostly consisted in 1–3 orthodromic spikes followed by an inhibition. During iontophoretic application of bicuculline, stimulation induced additional short latency orthodromic spikes, even in cells that were previously unresponsive. However, the duration of the inhibition was never reduced. These results indicate that GABAergic neurotransmission may play a dominant role in both spontaneous and evoked electrical activities in the CeL, probably mediated by local circuit cells involved in a feed-forward inhibition. This organization, along with the reciprocal connections between the CeL and the BSTL, is considered in the context of the extended amygdala.     

Intrinsic functional connectivity of the central extended amygdala

The central extended amygdala (EAc)—including the bed nucleus of the stria terminalis (BST) and central nucleus of the amygdala (Ce)—plays a critical role in triggering fear and anxiety and is implicated in the development of a range of debilitating neuropsychiatric disorders. Although it is widely believed that these disorders reflect the coordinated activity of distributed neural circuits, the functional architecture of the EAc network and the degree to which the BST and the Ce show distinct patterns of functional connectivity is unclear. Here, we used a novel combination of imaging approaches to trace the connectivity of the BST and the Ce in 130 healthy, racially diverse, community‐dwelling adults. Multiband imaging, high‐precision registration techniques, and spatially unsmoothed data maximized anatomical specificity. Using newly developed seed regions, whole‐brain regression analyses revealed robust functional connectivity between the BST and Ce via the sublenticular extended amygdala, the ribbon of subcortical gray matter encompassing the ventral amygdalofugal pathway. Both regions displayed coupling with the ventromedial prefrontal cortex (vmPFC), midcingulate cortex (MCC), insula, and anterior hippocampus. The BST showed stronger connectivity with the thalamus, striatum, periaqueductal gray, and several prefrontal territories. The only regions showing stronger functional connectivity with the Ce were neighboring regions of the dorsal amygdala, amygdalohippocampal area, and anterior hippocampus. These observations provide a baseline against which to compare a range of special populations, inform our understanding of the role of the EAc in normal and pathological fear and anxiety, and showcase image registration techniques that are likely to be useful for researchers working with “deidentified” neuroimaging data.     

Desipramine prevents the sustained increase in corticotropin-releasing hormone-like immunoreactivity induced by repeated immobilization stress in the rat central extended amygdala

Clinical and experimental studies have shown that the activation of corticotropin-releasing hormone (CRH) and noradrenergic systems mediate stress-induced anxiety. Repeated immobilization stress (RIS) has been shown to induce long-lasting anxiety behavior and changes in noradrenaline turnover. The present work was aimed at studying the effect of RIS on the in situ expression of CRH-LI in the central extended amygdala and paraventricular nucleus of the hypothalamus (PVN). Our results showed that RIS for 15 days induces a significant increase of CRH-LI expression in the central extended amygdala. The increase in CRH-LI expression in the central extended amygdala was sustained even after a 25-day stress-free period. The concomitant administration of desipramine (DMI), a specific noradrenaline uptake inhibitor, fully prevented the RIS-induced increase in CRH expression. RIS also induced an increase of CRH-LI expression in the PVN that was prevented by the concomitant DMI administration. In contrast to the sustained effect observed in the central extended amygdala, the RIS-induced increase of CRH-LI expression in the PVN was nonlasting. DMI administration also prevented the RIS-induced increase of adrenal gland weight. The present findings showing that RIS induces a sustained increase of CRH expression in the central extended amygdala suggest that the repeated activation of CRH neurons and CRH receptors in the central extended amygdala may underlie the long-lasting anxiety behavior induced by RIS. Further studies should address the mechanisms involved in the effect of DMI and its eventual relevance in the therapeutic actions of DMI.     

Origin of the dopaminergic innervation of the central extended amygdala and accumbens shell: a combined retrograde tracing and immunohistochemical study in the rat

The origin of the dopaminergic innervation of the central extended amygdala (EAc; i.e., the lateral bed nucleus of the stria terminalis [BSTl]-central amygdaloid nucleus [Ce] continuum) and accumbens shell (AcSh) was studied in the rat by combining retrograde transport of Fluoro-Gold (FG) with tyrosine hydroxylase (TH) immunofluorescence. Perikaryal profiles (PP) immunoreactive to FG and to both FG and TH were counted in A8-A14 dopaminergic districts. Our results suggest that dopaminergic inputs to the EAc and AcSh arise from the ventral tegmental area-A10, substantia nigra, pars compacta-A9, and retrorubral nucleus-A8 groups as well as from the dorsal raphe nucleus and periaqueductal gray substance, housing the dorsocaudal part of A10 group (A10dc). Quantitative estimates reveal that the A10dc group contains approximately half of the total number of FG/TH double-labeled PP projecting to Ce and BSTl. By using an anti-dopamine serum, DR/PAG projections to Ce were confirmed to be in part dopaminergic. In contrast, modest numbers of FG/TH double-labeled PP were seen in the A10dc group after injections in the sublenticular extended amygdala, interstitial nucleus of the posterior limb of the anterior commissure or AcSh. Ventral mesencephalic projections to the EAc display a crude mediolateral topographic organization, whereas those to the AcSh are topographically organized along a mediolateral and an inverted dorsoventral dimension. The diencephalic dopaminergic groups do not innervate the EAc or AcSh, except for the periventricular gray-A11 which sends light dopaminergic projections to Ce and BSTl. Overall, the present results provide additional details on the organization of the mesolimbic dopaminergic system that critically controls behavioral responsiveness to salient environmental stimuli.     

Projections from the nociceptive area of the central nucleus of the amygdala to the forebrain: a PHA-L study in the rat

The lateral capsular division (CeLC) of the central nucleus (Ce) of the amygdala, in the rat, has been shown to be the main terminal area of a spino(trigemino)-parabrachio-amygdaloid nociceptive pathway [Bernard & Besson (1990) J. Neurophysiol. 63, 473-490; Bernard et al. (1992) J. Neurophysiol. 68, 551-569; Bernard et al. (1993) J. Comp. Neurol. 329, 201-229]. The projections to the forebrain from the CeLC and adjacent regions were studied in the rat by using microinjections of Phaseolus vulgaris leucoagglutinin (PHA-L) restricted in subdivisions of the Ce and the basolateral amygdaloid nucleus anterior (BLA). Our data showed that the entire CeLC projects primarily and extensively to the substantia innominata dorsalis (SId). The terminal labelling is especially dense in the caudal aspect of the SId. The other projections of the CeLC in the forebrain were dramatically less dense. They terminate in the bed nucleus of the stria terminalis (BST) and the posterior hypothalamus (pLH). No (or only scarce) other projections were found in the remaining forebrain areas. The Ce lateral division (CeL) and the Ce medial division (CeM), adjacent to the CeLC, also project to the SId with slightly lower density labelling. However, contrary to the case of the CeLC, both the CeL and the CeM extensively project to the ventrolateral subnucleus of the BST (BSTvl) with a few additional terminals found in other regions of the lateral BST. Only the CeM projects densely to both the interstitial nucleus of the posterior limb of the anterior commissure and the caudal most portion of the pLH. The projections of the BLA are totally different from those of the Ce as they terminate in the dorsal striatum, the accumbens nucleus, the olfactory tubercle, the nucleus of olfactory tract and the rostral pole of the cingulate/frontal cortex. This study demonstrates that the major output of the nociceptive spino(trigemino)-parabrachio-CeLC pathway is to the SId. It is suggested that the CeLC-SId pathway could have an important role in anxiety, aversion and genesis of fear in response to noxious stimuli.     

Organization of multisynaptic circuits within and between the medial and the central extended amygdala

The central and medial extended amygdala comprises the central (CEA) and medial nuclei of the amygdala (MEA), respectively, together with anatomically connected regions of the bed nucleus of the stria terminalis (BST). To reveal direct and multisynaptic connections within the central and medial extended amygdala, monosynaptic and transneuronal viral tracing experiments were performed in adult male rats. In the first set of experiments, a cocktail of anterograde and retrograde tracers was iontophoretically delivered into the medial CEA (CEAm), anterodorsal MEA (MEAad), or posterodorsal MEA (MEApd), revealing direct, topographically organized projections between distinct amygdalar and BST subnuclei. In the second set of experiments, the retrograde transneuronal tracer pseudorabies virus (PRV) was microinjected into the CEAm or MEAad. After 48 hours of survival, there were no significant differences between monosynaptic and PRV cases in the subnuclear distribution or proportions of retrogradely labeled BST neurons. However, after 60 hours of survival, CEAm‐injected cases displayed an increased proportion of labeled neurons within the anteromedial group of BST subnuclei (amgBST) and within the posterior BST, which do not directly innervate the CEA. MEApd‐injected 60‐hour cases displayed a significantly increased proportion of retrograde labeling in the amgBST compared with monosynaptic and 48‐hour cases, whereas MEAad‐injected cases displayed no proportional changes over time. Thus, multisynaptic circuits within the medial extended amygdala overlap the direct connections making up this anatomical unit, whereas the multisynaptic boundaries of the central extended amygdala extend into BST subnuclei previously identified as part of the medial extended amygdala. J. Comp. Neurol. 521:3406‐3431, 2013. © 2013 Wiley Periodicals, Inc.     

c-Fos and peptide immunoreactivities in the central extended amygdala of morphine-dependent rats after naloxone-precipitated withdrawal

The central extended amygdala, a forebrain macrostructure, may represent a common substrate for acute drug reward and the dysphoric effects of drug withdrawal. To test its involvement during opiate withdrawal, we studied the distribution of c-Fos immunoreactive neurons, in relation to their neuropeptide content, in brain sections from morphine-dependent or naive rats, killed 90 min after naloxone or saline intraperitoneal injection. Naloxone treatment in naive rats induced a slight increase in c-Fos immunoreactivity in the central amygdaloid nucleus, the lateral bed nucleus of the stria terminalis and the interstitial nucleus of the posterior limb of the anterior commissure. In morphine-dependent rats, naloxone injection significantly increased the number of c-Fos-positive neurons in these structures as well as in the majority of the other central extended amygdala components. Double immunocytochemistry was used to determine the neurochemical nature of c-Fos-positive neurons in the central extended amygdala. Corticotropin-releasing factor- and methionine-enkephakin-immunoreactive neurons displayed c-Fos immunoreactivity in naive rats after naloxone injection, whereas only enkephalinergic neurons were found to be c-Fos positive in morphine-dependent rats after naloxone injection. The possible involvement of the corticotropin-releasing factor system during withdrawal is discussed. These results suggest that the whole central extended amygdala is activated during opiate withdrawal, with a lateral to medial decreasing gradient, and emphasize the role of peptidergic systems in this morphofunctional continuum.     

Branching patterns of parabrachial neurons projecting to the central extended amgydala: single axonal reconstructions

Electrophysiological evidence suggests that the spinoparabrachioamygdaloid pathway carries nociceptive information that may be important for the elaboration of physiological and emotional responses to noxious events. The pontine parabrachial nucleus (pPB) sends a massive projection to the central nucleus of the amygdala (CeA) and lateral bed nucleus of the stria terminalis (BSTL), both regions belonging to a broader macrostructure, the central extended amygdala (EAc). The aim of this study was to examine whether different EAc components are targeted by a same pPB neuron, by reconstructing single axonal branching patterns after anterograde labelling. Small deposits of biotinylated dextran amine in the region of the external lateral pPB result in dense and specific labelling in the whole EAc. Reconstructed axons innervate either the lateral or the capsular part of the CeA with perisomatic or bushy terminals, respectively. A subset of axons enters the stria terminalis rostrally to follow its trajectory caudally toward the CeA. Individual axons targeting the CeA usually send collaterals to other EAc components, especially those projecting to the lateral CeA, which often coinnervate the BSTL. By contrast, only few branches were found outside the EAc. These results suggest that the noxious information travelling from the pPB to the CeA may also be transmitted to other EAc components. This pPB-EAc pathway, which appears distinct from the parabrachiohypothalamic and parabrachiothalamic projections, would be the anatomical basis through which the EAc elaborates the autonomic, endocrine, and emotional components of pain.     

Sex-dependent and differential responses to acute restraint stress of corticotropin-releasing factor-producing neurons in the rat paraventricular nucleus, central amygdala, and bed nucleus of the stria terminalis

Male and female rodents respond differently to acute stress. We tested our hypothesis that this sex difference is based on differences in stress sensitivity of forebrain areas, by determining possible effects of a single acute psychogenic stressor (1-hr restraint stress) on neuronal gene expression (c-Fos and FosB immunoreactivities), storage of corticotropin-releasing factor (CRF) immunoreactivity, and CRF production (CRF mRNA in situ hybridization) as well as the expression of genes associated with epigenetic processes (quantitative RT-PCR) in the rat paraventricular nucleus (PVN), the oval and fusiform subdivisions of the bed nucleus of the stria terminalis (BSTov and BSTfu, respectively), and the central amygdala (CeA), in both males and females. Compared with females, male rats responded to the stressor with a stronger rise in corticosterone titer and a stronger increase in neuronal contents of c-Fos, CRF mRNA, and CREB-binding protein mRNA in the PVN. In the BSTov, females but not males showed an increase in c-Fos, whereas the CRF mRNA content was increased in males only. In the BSTfu, males and females showed similar stress-induced increases in c-Fos and FosB, whereas in the CeA, both sexes revealed similar increases in c-Fos and in CRF mRNA. We conclude that male and female rats differ in their reactivity to acute stress with respect to possibly epigenetically mediated (particularly in the PVN) neuronal gene expression and neuropeptide dynamics (PVN and BSTov) and that this difference may contribute to the sex dependence of the animal's physiological and behavioral responses to an acute stressor.     

Distribution of pituitary adenylate cyclase activating polypeptide (PACAP) immunoreactivity in the hypothalamus and extended amygdala of the rat

Pituitary adenylate cyclase activating polypeptide (PACAP) is found in two forms of 27 and 38 amino acids (PACAP-27 and PACAP-38 respectively) in the mammalian central nervous system. Using antibodies to these two forms of PACAP, we examined the distribution of PACAP immunoreactivity in the rat hypothalamus and a number of extrahypothalamic areas. The patterns of immunostaining for PACAP-27 and PACAP-38 were similar: prominent terminal labelling was present in the retrochiasmatic area, median eminence, and posterior periventricular nucleus of the hypothalamus as well as the bed nucleus of the stria terminalis and amygdaloid complex. After colchicine treatment, immunopositive cell bodies were found in the preoptic region of the periventricular zone of the hypothalamus, the suprachiasmatic and paraventricular hypothalamic nuclei, neural structures adjacent to the median eminence (including the retrochiasmatic area, arcuate nucleus, ventromedial hypothalamus, and tuber cinereum), and the lateral mammillary and supramammillary nuclei. In all these areas, immunolabelling appeared specific since it was abolished by preabsorption of primary antisera with the appropriate PACAP peptide. However, the number of immunopositive cells in the suprachiasmatic nucleus was also reduced by preabsorption of PACAP-27/38 antisera with vasoactive intestinal polypeptide, suggesting that a subpopulation of cells in the suprachiasmatic nucleus express a peptide which has significant sequence homology with both PACAP-27/38 and vasoactive intestinal polypeptide. The distribution of PACAP immunoreactivity throughout the hypothalamus, bed nucleus of the stria terminalis, and amygdala suggests the involvement of PACAP in a number of processes including limbic, autonomic, and neuroendocrine functions as well as regulation of the circadian pacemaker. © 1996 Wiley-Liss, Inc.     

Projections from the paraventricular nucleus of the thalamus to the forebrain, with special emphasis on the extended amygdala

The paraventricular nucleus of the thalamus (PVT) is part of a group of midline and intralaminar thalamic nuclei implicated in arousal and attention. This study examined the connections between the PVT and the forebrain by using the retrograde tracer cholera toxin B (CTb) and the anterograde tracer biotin dextran amine (BDA). The anterior and posterior regions of the PVT were found to send a dense projection to the nucleus accumbens. The posterior PVT was also found to provide a strong projection to the lateral bed nucleus of the stria terminalis (BST), interstitial nucleus of the posterior limb of the anterior commissure (IPAC), and central nucleus of the amygdala (CeA), regions associated with the extended amygdala. In contrast, the anterior PVT was found to send a weaker projection to the extended amygdala. The basolateral nucleus of the amygdala and the medial prefrontal cortex were found to receive a relatively weak projection from the PVT, and other regions of the BST and amygdala were found to be poorly innervated by the PVT. In addition, the PVT was found to innervate regions in the extended amygdala that contained corticotropin-releasing factor (CRF) neurons, many of which were found to receive apparent contacts from PVT fibers. The projection from the PVT to the nucleus accumbens and extended amygdala places the PVT in a key anatomical position to influence adaptive behaviors as well as the physiological and neuroendocrine responses associated with these behaviors.     

Inhibition of the central extended amygdala by loud noise and restraint stress

It is well established that the central nucleus of the amygdala (CEA) is involved in responses to stress, fear and anxiety. Many studies have used c-fos expression to map the brain's response to processive stress, but curiously the CEA generally is not highly activated. We have previously shown that exposure to a novel vs. home environment reduces amphetamine-induced activation of the lateral CEA (CEAl) and the oval nucleus of the bed nucleus of the stria terminalis (BSTov). This is consistent with the idea that processive stress inhibits neurons in these nuclei. We have tested this hypothesis by exposing rats to noise, at a range of intensities from non-stressful to stressful, or to restraint conditions, immediately after a remote injection of amphetamine, 2 mg/kg i.p., or interleukin-1beta (IL-1beta) 0.5 microg/kg i.p. (used to obtain a level of c-fos mRNA against which to measure inhibition). In keeping with our hypothesis, amphetamine- or IL-1beta-induced c-fos and zif-268 mRNA were significantly decreased in the CEAl and BSTov under conditions of loud noise or restraint stress compared with control conditions. This inhibition does not require a stress-induced rise in corticosterone because data were similar in animals that had been adrenalectomized with a low-dose corticosterone replacement. As both the CEAl and BSTov are highly gamma-aminobutyric acid (GABA) -ergic and project to the medial CEA (CEAm), their inhibition potentially causes an increased input to the CEAm. As the CEAm is a major output nucleus of the amygdala, this could have important consequences within the neural circuitry controlling responses to processive stress.     

Effects of chronic stress on dendritic arborization in the central and extended amygdala

A differential role has been suggested for two important areas in the neural circuitry of stress, central nucleus of the amygdala (CeA) and bed nucleus of stria terminalis (BNST) in the extended amygdala, in regulating fear versus anxiety. Since chronic stress enhances anxiety and consolidation of aversive memories, we examined the effects of chronic immobilization stress (CIS) on neuronal morphology in the CeA and BNST of rats. In contrast to previous reports of stress-induced atrophy in the hippocampus, CIS does not cause dendritic atrophy in CeA and BNST neurons. While dendritic arborization in CeA neurons remains unaffected, it increases in BNST neurons after CIS. These results suggest a role for dendritic remodeling of BNST neurons in stress-induced facilitation of anxiety.     

Enkephalinergic afferents of the centromedial amygdala in the rat

The connectivity of the amygdaloid complex has been extensively explored with both anterograde and retrograde tracers. Even though the afferents of the centromedial amygdala [comprising the central (CEA) and medial (MEA) amygdaloid nuclei] are well established, relatively little is known about the neuropeptide phenotype of these connections. In this study, we first examined the distribution of mu-opioid receptor (MOR) and delta-opioid receptor (DOR) in the amygdala via in situ hybridization and immunohistochemistry. We then investigated the distribution of Met-enkephalin (ENK) and Leu-ENK fibers with immunohistochemistry and examined the distribution of preproenkephalin mRNA in the amygdala by using in situ hybridization. Finally, we examined the ENK projections to the CEA and MEA by using stereotaxic injections of the retrograde tracer cholera toxin subunit B or fluorogold revealed by immunohistochemistry combined with in situ hybridization to identify ENKergic neurons. Our results indicate that the centromedial amygdala receives ENK afferents, as indicated by the presence of MOR, DOR, and ENK fibers in the CEA and MEA, originating primarily from the bed nucleus of the stria terminalis (BST) and from other amygdaloid nuclei. The posterior BST, the basomedial nucleus (BMA), and the cortical nucleus of the amygdala (COA) were found to be the major ENK afferents of the MEA, whereas the anterolateral BST, the COA, the MEA, and the BMA provided the main ENKergic innervation of the CEA. In addition, we found that the ventromedial nucleus of the hypothalamus and the pontine parabrachial nucleus provide a moderate ENK input to the CEA and MEA. The functional implications of these connections in stress, anxiety, and nociception are discussed.     

Antinociception after microinjection of neurotensin into the central amygdaloid nucleus of the rat

Neurotensin (NT) is an endogenous peptide which has been hypothesized to function in the central nervous systems as a neurotransmitter. Injection of NT into the cerebral ventricular system of rodents produces antinociception in a variety of analgesia tests. In the hot plate test, direct microinjection of NT into the central nucleus of the amygdala (AC) produced a significant increase in the nociceptive threshold of the rat, while injections into tissue adjacent to the AC were generally ineffective. Antinociception following intra-AC injection of NT occurred at an ED₅₀ dose of 2.4 μg NT, and was significantly lower than the ED₅₀ dose observed when NT was given into the lateral ventricles (93.2 μg NT). Lesions of the stria terminalis totally abolished the antinociceptive effect of intra-AC administration of NT, indicating that AC efferent or afferent fibers within the stria terminalis are necessary for the observed increase in nociceptive threshold.     

Pattern of distribution of serotonergic fibers to the amygdala and extended amygdala in the rat

As is well recognized, serotonergic (5‐HT) fibers distribute widely throughout the forebrain, including the amygdala. Although a few reports have examined the 5‐HT innervation of select nuclei of the amygdala in the rat, no previous report has described overall 5‐HT projections to the amygdala in the rat. Using immunostaining for the serotonin transporter, SERT, we describe the complete pattern of distribution of 5‐HT fibers to the amygdala (proper) and to the extended amygdala in the rat. Based on its ontogenetic origins, the amygdala was subdivided into two major parts, pallial and subpallial components, with the pallial component further divided into superficial and deep nuclei (Olucha‐Bordonau et al. 2015). SERT⁺ fibers were shown to distributed moderately to densely to the deep and cortical pallial nuclei, but, by contrast, lightly to the subpallial nuclei. Specifically, 1) of the deep pallial nuclei, the lateral, basolateral, and basomedial nuclei contained a very dense concentration of 5‐HT fibers; 2) of the cortical pallial nuclei, the anterior cortical and amygdala–cortical transition zone rostrally and the posteromedial and posterolateral nuclei caudally contained a moderate concentration of 5‐HT fibers; and 3) of the subpallial nuclei, the anterior nuclei and the rostral part of the medial (Me) nuclei contained a moderate concentration of 5‐HT fibers, whereas caudal regions of Me as well as the central nuclei and the intercalated nuclei contained a sparse/light concentration of 5‐HT fibers. With regard to the extended amygdala (primarily the bed nucleus of stria terminalis; BST), on the whole, the BST contained moderate numbers of 5‐HT fibers, spread fairly uniformly throughout BST. The findings are discussed with respect to a critical serotonergic influence on the amygdala, particularly on the basal complex, and on the extended amygdala in the control of states of fear and anxiety. J. Comp. Neurol. 525:116–139, 2017. © 2016 Wiley Periodicals, Inc.     

Fiber connections of the compact division of the posterior pallial amygdala and lateral part of the bed nucleus of the stria terminalis in the pigeon (Columba livia)

The compact division of the posterior pallial amygdala (PoAc) and lateral part of the bed nucleus of the stria terminalis (BSTL) are components of the limbic system in the pigeon brain. In this study, we examined the position and fiber connections of these two nuclei by using Nissl staining and tract-tracing methods. PoAc occupies a central division in the posterior pallial amygdala. BSTL faces the ventral horn of the lateral ventricle and extends between A 7.25 and A 10.50. PoAc and BSTL connect bidirectionally by the stria terminalis. PoAc connects reciprocally with two nuclear groups in the cerebrum: 1) a continuum consisting of the caudoventral nidopallium, lateral part of the caudoventral nidopallium (NCVl), subnidopallium beneath NCVl, and piriform cortex and 2) rostral areas of the hemisphere, including the frontolateral and frontomedial nidopallium and the densocellular part of the hyperpallium. Extratelencephalic projections of PoAc terminate in the dorsomedial thalamic nuclei and reach the lateral hypothalamic area via the hypothalamic part of the occipito-mesencephalic tract. BSTL also connects reciprocally with two main regions: 1) the same continuum as for PoAc projections, except the piriform cortex and 2) rostral areas of the hemisphere, including the olfactory tubercle and nucleus accumbens. Extratelencephalic reciprocal connections are with the substantia nigra, nucleus subceruleus dorsalis, parabrachial nucleus, locus coeruleus, and nucleus of the solitary tract. The dorsomedial subdivision of the hippocampal formation projects massively to PoAc and BSTL. These findings indicate that PoAc and BSTL are important components of an interconnected neural circuit involving widespread regions of the neuraxis and mediating limbic-visceral functions.