The distribution of GABA-containing perikarya,fibers, and terminals in the forebrain and midbrain of pigeons,with particular reference to the basal ganglia and its projection targets

Immunohistochemical techniques were used to study the distributions of glutamic acid decarboxylase (GAD) and γ-aminobutyric acid (GABA) in pigeon forebrain and midbrain to determine the organization of GABAergic systems in these brain areas in birds. In the basal ganglia, numerous medium-sized neurons throughout the striatum were labeled for GABA, while pallidal neurons, as well as a small population of large, aspiny striatal neurons, labeled for GAD and GABA. GAD+ and GABA+ fibers and terminals were abundant throughout the basal ganglia, and GABAergic fibers were found in all extratelencephalic targets of the basal ganglia. Most of these targets also contained numerous GABAergic neurons. In pallial regions, approximately 10-12% of the neurons were GABAergic. The outer rind of the pallium was more intensely labeled for GABAergic fibers than the core. The olfactory tubercle region, the ventral pallidum, and the hypothalamus were extremely densely labeled for GABAergic fibers, while GABAergic neurons were unevenly distributed in the hypothalamus. GABAergic neurons and fibers were abundant in the dorsalmost part of thalamus and the dorsal geniculate region, while GABAergic neurons and fibers were sparse (or lightly labeled) in the thalamic nuclei rotundus, triangularis, and ovoidalis. Further, GABAergic neurons were abundant in the superficial tectal layers, the magnocellular isthmic nucleus, the inferior colliculus, the intercollicular region, the central gray, and the reticular formation. GABAergic fibers were particularly abundant in the superficial tectal layers, the parvocellular isthmic nucleus, the inferior colliculus, the intercol-licular region, the central gray, and the interpeduncular nucleus. These results suggest that GABA plays a role as a neurotransmitter in nearly all fore- and midbrain regions of birds, and in many instances the observed distributions of GABAergic neurons and fibers closely resemble the patterns seen in mammals, as well as in other vertebrates. © 1994 Wiley-Liss, Inc.     

Localization of GABA immunoreactivity in the auditory brainstem of guinea pigs

Using an antibody against GABA, neurons within the guinea pig hindbrain, midbrain and forebrain auditory nuclei were identified which demonstrate GABA-like immunoreactivity. GABA-positive cells were localized in the cochlear nucleus, superior olivary complex, lateral lemniscus, inferior colliculus, and medial geniculate body. GABA-positive terminals could be seen surrounding globular and spherical cells in ventral cochlear nucleus and principal cells in medial nucleus of the trapezoid body. In addition, numerous positive, punctate terminals appeared throughout the hindbrain auditory nuclei and, although fewer in number, in midbrain and forebrain auditory nuclei.     

Distribution of GABAergic neurons and terminals in the auditory system of the barn owl

Antisera to GAD (glutamic acid decarboxylase) and GABA were used to determine the distribution of GABAergic cells and terminals in the brainstem and midbrain auditory nuclei of the barn owl. The owl processes time and intensity components of the auditory signal in separate pathways, and each pathway has a distinctive pattern of GAD- and GABA-like immunoreactivity. In the time pathway, all the cells of the cochlear nucleus magnocellularis and nucleus laminaris receive perisomatic GABAergic terminals, and small numbers of GABAergic neurons surround both nuclei. The ventral nucleus of the lateral lemniscus (anterior division) contains both immunoreactive terminals and some GABAergic neurons. In the intensity pathway, dense immunoreactive terminals are distributed throughout the cochlear nucleus angularis, which also contains a small number of GABAergic neurons. The superior olive contains two GABAergic cell types and immunoreactive terminals distributed throughout the neuropil. All the neurons of the nucleus of the lateral lemniscus (ventral part) appear to be GABAergic, and this nucleus also contains a moderate number of immunoreactive terminals. Immunoreactive terminals are distributed throughout the neuropil of the ventral nucleus of the lateral lemniscus (posterior division), whereas multipolar and small fusiform GABAergic neurons predominate in the dorsal regions of the nucleus. The time and intensity pathways combine in the inferior colliculus. The central nucleus of the inferior colliculus contains a larger number of fusiform and stellate GABAergic neurons and a dense plexus of immunoreactive terminals, whereas the external nucleus contains slightly fewer immunoreactive cells and terminals. The superficial nucleus contains dense, fine immunoreactive terminals and a small number of GABAergic neurons.     

Neuropeptide Y in the cerebral cortex and the caudate-putamen nuclei: ultrastructural basis for interactions with GABAergic and non-GABAergic neurons

In the cerebral cortex and caudate-putamen (CP) nuclei, neuropeptide Y (NPY) immunoreactivity is detectable within 1-2% of all neurons. The NPY-immunoreactive neurons are interneuronal and are believed to be mostly GABAergic in the cerebral cortex but not in the CP nuclei. Thus NPY and GABA may play different roles in the circuitry within these 2 regions. We tested this possibility by comparing the ultrastructure of NPY-containing neurons between (1) cortex (somatosensory and anterior cingulate areas) versus dorsolateral CP; and (2) GABAergic versus non-GABAergic NPY neurons within each area. Single coronal sections through the rat forebrain were dually labeled for GABA and NPY by combining immunoautoradiography with the immunoperoxidase method. NPY-containing neurons with or without GABA occurred throughout the rostrocaudal portions of CP and all laminae of somatosensory and anterior cingulate cortex. Comparisons between the areas confirmed that somata and terminals dually labeled for GABA and NPY were more prevalent in the cortex. NPY terminals lacking detectable GABA immunoreactivity also were found within the cortex, thus suggesting additional heterogeneity in cortical NPY innervation. The ultrastructural features of NPY perikarya in both regions were morphologically similar regardless of whether the cells also contained GABA. Most synaptic inputs to NPY neurons occurred at distal dendrites. In comparison to neighboring neurons, synaptic inputs to proximal dendrites and somata of NPY neurons of cortex and CP were rare, suggesting that fewer and weaker inputs may modulate the excitability of NPY-containing neurons. In both regions, nearly all NPY- and NPY-GABA-labeled terminals formed symmetric junctions suggestive of inhibitory action. The majority of these junctions were with dendrites containing neither NPY nor GABA. NPY terminals formed few contacts on proximal dendrites and somata of GABAergic neurons (8% of 179 contacts in cortex; 12% of 73 contacts in CP) which, unlike most singly-labeled GABAergic neurons, were sparsely innervated. Thus, NPY may play a more prominent role in modulation of certain GABAergic neurons than would be predicted by the observed frequency of NPY-to-GABA contacts in the two regions. One notable regional difference was the greater prevalence in cortex of axoaxonic associations between NPY-immunoreactive terminals and other terminals, some of which also contained NPY. These nonsynaptic associations may be involved in the modulation of (1) the release of NPY by another transmitter or (2) NPY's modulation of release of other transmitters in cortex.     

GABA-immunoreactive neurons and terminals in the lateral cervical nucleus of the cynomolgus monkey

An antiserum against the inhibitory transmitter substance gamma-aminobutyric acid (GABA) was used to investigate the distribution of GABAergic nerve terminals and cell bodies in the lateral cervical nucleus (LCN) of the cynomolgus monkey. Light microscopic immunohistochemistry demonstrated GABA-immunoreactive puncta, suggestive of nerve terminals, scattered throughout the LCN. The terminal-like profiles are often present along the somata of unlabeled neurons, but most are located in the neuropil. GABA-immunoreactive neurons are present in the LCN, but constitute a very small number of the LCN neurons. Electron microscopy showed that the GABA-positive neurons are small with a relatively large nucleus. They are contacted by few somatic boutons. Numerous GABA-immunoreactive terminals containing densely packed round to oval synaptic vesicles were also found. Most GABA-positive terminals make synaptic contact with dendrites, but synapses with cell bodies are also present. Synaptic contacts between labeled and unlabeled terminals were not observed. Some GABA-positive terminals make contact with GABA-positive neurons. The present findings suggest that GABA is a major inhibitory transmitter substance in the LCN of the monkey. However, in comparison with other somatosensory relay nuclei, there are few GABA-immunoreactive neurons in the LCN. This may imply that the GABA-positive neurons branch extensively in the LCN or that an extrinsic source of GABAergic input exists.     

Immunocytochemical evidence for the existence of GABAergic neurons in the nucleus raphe dorsalis. possible existence of neurons containing serotonin and GABA

It has been established that nerve cell bodies of the nucleus raphe dorsalis (NRD) belong to ascending 5-hydroxytryptamine systems. These neurons could be modulated by GABAergic interneurons or interposed GABA neurons. A high glutamate decar☐ylase (GAD) activity in the NRD and a specific high-affinity uptake mechanism for GABA suggest the presence of GABA synthesizing elements in the NRD. Anti-GAD antibodies were used by an immunocytochemical procedure to demonstrate the presence of GABAergic elements. Anti-GAD antibodies were previously tested in the cerebellum and substantia nigra. Large amounts of GAD-positive reaction product were observed in the cytoplasm of some neurons (fusiform, ovoid or multipolar) or appeared as punctate deposits apposed to dendrites, soma and dispersed in the neuropil of the NRD. At the electron microscopic level, GAD-positive reaction product was observed within the cytoplasm of numerous somata in sections from colchicine-treated rats. GAD-positive staining was observed in numerous fibers or axonal terminals and two types of morphologically different fibers could be distinguished. The first displays small clear vesicles and few large granular vesicles (LGV) (80–100 nm), the second displays only clear round vesicles (40–60 nm). After 5,7-dihydroxytryptamine treatment (a neurotoxic for 5-HT terminals), the immunocytochemical labeling is much decreased. Some reactive neurons are still dispersed in the nucleus but the fibers containing LGV are no longer observed. These results strongly suggest that some neuronal elements in the NRD are morphologically, pharmacologically and anatomically similar to 5-HT neurons described at this level. Such cell elements could possess a double GABA and 5-HT potentiality. If this is not the case, a population of GABA neurons could be sensitive to 5,7-DHT and so have the capacity to take up 5-HT. The other reactive elements, insensitive to 5,7-DHT, could represent the GABAergic interneurons postulated at this level. Numerous GAD positive fibers or axon terminals were observed in synaptic contact with dendrites, axons or soma of other neurons. The chemical nature of the neuronal postsynaptic elements remains unknown. These findings strongly support the hypothesis for GABA-mediated inhibition in the NRD.     

Light and electron microscopic evidence for a GABAergic projection from the caudal basal forebrain to the thalamic reticular nucleus in rats.

Neurons in the magnocellular nucleus of the caudal basal forebrain extend an axonal projection which arborizes within the reticular nucleus of the thalamus. The present study addresses the ultrastructure and neurochemistry of this projection in rats. Many labeled terminals are apparent within the thalamic reticular nucleus following Phaseolus vulgaris leucoagglutinin injections into the caudal basal nucleus; anterogradely labeled axon terminals most commonly contact both somata and dendrites of reticular nucleus neurons with symmetric membrane specializations. Thus, the majority of the labeled terminals examined contrast with choline acetyltransferase positive terminals which have been previously identified as contacting dendrites and forming asymmetric synapses within this nucleus. Many of the neurons within the caudal basal nucleus which are retrogradely labeled following tracer injections into the thalamic reticular nucleus are gamma-aminobutyric acid (GABA) immunoreactive. In addition, following injections of Phaseolus vulgaris leucoagglutinin or fluoro-ruby into the caudal basal forebrain, some of the labeled axonal swellings and boutons within the thalamic reticular nucleus also contain glutamic acid decarboxylase. These results indicate that a significant component of the projection is GABAergic. These anatomical observations suggest that the projection from the caudal basal nucleus onto the thalamic reticular nucleus could facilitate the relay of information through the dorsal thalamus by inhibiting reticular nucleus neurons, and thus, in turn, disinhibiting thalamic relay neurons.     

Relationship of opioid receptors with GABAergic neurons in the rat inferior colliculus

The inferior colliculus is a critical structure for processing auditory information and receives ascending and descending synaptic auditory projections. In addition to GABAergic and glutamatergic innervations, other neurotransmitter systems are also reported in the inferior colliculus, including opioid peptides. In the present study, the relative distribution of each type of opioid receptor, mu (MOR), delta (DOR) and kappa (KOR) within GABAergic neurons in the inferior colliculus was examined. GABA immunoreactivity was expressed by small, medium and large neurons and distributed in the central nucleus and the pericentral nucleus of the inferior colliculus. Immunostaining for MOR, DOR and KOR receptors was found in both disc-shaped cells and stellate cells. Punctiform beta-endorphin immunolabelling was observed in the proximity of GABA-positive neurons. Co-localization of GABA and MOR receptors was observed in neurons and nerve terminals in the central nucleus, dorsal cortex and external cortex of the inferior colliculus. Quantification of the co-localization patterns determined that a higher proportion of GABA neurons was associated with MOR receptors compared with KOR or DOR receptors.     

Ultrastructural organization of the retino-pretecto-olivary pathway in the rabbit: a combined WGA-HRP tracing and GABA immunocytochemical study.

The ultrastructural organization of the pretecto-olivary projection neurons within the nucleus of the optic tract and dorsal terminal accessory optic nucleus of rabbits was studied by using anti-GABA immunolabelling and retrograde transport of WGA-HRP. GABA-like immunoreactivity was determined with a postembedding colloidal gold technique. WGA-HRP was injected in the dorsal cap of the inferior olive. The WGA-HRP-labelled neurons were incubated with gold-substituted silver peroxidase. Neurons projecting to the inferior olive had large to medium-sized cell bodies and were GABA negative. In the nucleus of the optic tract, projection neurons are found in the rostral parts, while the majority of the local GABAergic interneurons are mainly found in the caudal parts. In the dorsal terminal nucleus both types of neurons are intermingled. The projection neurons were frequently in synaptic contact by GABAergic terminals. These neurons also receive retinal afferents indicating the existence of a two-step synaptic connection from the retina to the inferior olive. It is suggested that this class of projection neurons forms the "direction-selective" neurons that can be antidromically stimulated from the inferior olive. The GABAergic terminals on the identified projection neurons are of axonal origin (F-terminals), whereas presynaptic dendrites of interneurons (P-terminals) were seldom observed to be in synaptic contact with retrogradely labelled profiles. The strong input of GABA on direction-selective neurons indicates that GABA is directly involved in modulating retinal signals to the inferior olive.     

Synaptic interactions between GABAergic neurons and trigeminothalamic cells in the rat trigeminal nucleus caudalis.

Synaptic interactions between GABAergic neurons and thalamic projecting cells within the trigeminal nucleus caudalis were examined using a combined method of GABA immunohistochemistry and retrograde WGA-HRP labeling of the trigeminothalamic pathway. Results showed that GABA-positive neurons and projecting cells were separate but closely intermingled within the spinal trigeminal nucleus. GABAergic axon terminals formed symmetric synaptic connections with the cell bodies and dendrites of trigeminothalamic neurons. In turn, some WGA-HRP filled axon terminals, presumed to originate from axon collaterals of the projecting neurons, formed synaptic connections with GABA containing neurons. The results suggest that in the spinal trigeminal nucleus there is a reciprocal modulation between GABA neurons and trigeminothalamic cells.     

In the ventromedial nucleus of the rat hypothalamus, GABA-immunolabeled neurons are abundant and are innervated by both enkephalin- and GABA-immunolabeled axon terminals

Immunohistochemical-labeling for the neurochemicals γ-aminobutyric acid (GABA) and enkephalin are abundant in the ventromedial nucleus of the hypothalamus (VMN). In VMN, both GABA and enkephalin may function to regulate feeding behavior, as well as other hormone-controlled behaviors. Importantly, in several brain areas, enkephalin is often thought to modulate GABAergic neurotransmission. Therefore, we used dual-labeling immunohistochemistry with electron microscopic analysis to study the circuitry of neurons containing GABA- and/or enkephalin-labeling within the VMN. Somato-dendritic profiles containing GABA-labeling were three fold more abundant than GABA-labeled axon terminals (117 soma or dendrites vs. 34 axons). In addition, axon terminals containing GABA-labeling sometimes synapsed onto GABA-labeled somata or dendrites (25% or 9/34). In contrast, under these conditions labeling for enkephalin was primarily restricted to axon terminals, which were very abundant throughout VMN. Enkephalin-containing terminals accounted for a large fraction (25% 23/92) of the axons in contact with GABA-labeled dendrites, although they also contacted unlabeled dendrites. These observations suggest that a population of VMN neurons are GABAergic. These may be either local circuit ‘interneurons' or projection neurons. In addition, GABA-labeled VMN neurons may be regulated by either enkephalin or GABA. These morphologic observations provide the basis for disinhibitory mechanisms to function within the VMN.     

Parvalbumin immunoreactivity in the thalamus of guinea pig: Light and electron microscopic correlation with gamma-aminobutyric acid immunoreactivity

The relationship of the calcium binding protein parvalbumin (PV) with gamma-aminobutyric acidergic (GABAergic) neurons differs within different thalamic nuclei and animal species. In this study, the distribution of PV and GABA throughout the thalamus of the guinea pig was investigated at the light microscopic level by using immunoperoxidase methods. Intense PV labelling was found in all the GABAergic neurons of the reticular nucleus and in scattered GABAergic neurons in the anteroventral nucleus, whereas GABAergic interneurons in the ventrobasal and lateral geniculate nuclei were not PV labelled. At the electron microscopic level, preembedding immunuperoxidase for PV was combined with postembedding immunogold for GABA. In the ventrobasal nucleus, four types of profiles were recognized: 1) terminals with flattened vesicles and forming symmetric synapses, which were labelled with both PV and GABA and could therefore be identified as afferents from the reticular nucleus; 2) boutons morphologically similar to presynaptic dendrites of interneurons, labelled only with GABA; 3) large terminals with round vesicles and asymmetric synapses, labelled only with PV, which contacted GABAergic presynaptic dendrites in glomerular arrangements and resembled ascending excitatory afferents; and 4) terminals unlabelled by either antiserum. In the ventrobasal nucleus of the guinea pig a double immunocytochemical labelling permits therefore the differentiation of two populations of GABAergic vesicle-containing profiles, i. e., the terminals originating from reticular nucleus (that are double labelled) and the presynaptic dendrites originating from interneurons (that are GABA-labelled only). The possibility to differentiate GABAergic inputs from the reticular nucleus and from interneurons can shed light to the functional interpretation of synaptic circuits in thalamic sensory nuclei. © 1994 Wiley-Liss, Inc.     

Contribution of brainstem GABAergic circuitry to descending antinociceptive controls: II. Electron microscopic immunocytochemical evidence of GABAergic control over the projection from the periaqueductal gray to the nucleus raphe magnus in the rat

Pharmacological, physiological, and behavioral studies suggest that inhibitory GABAergic neurons influence the projection from the midbrain periaqueductal gray matter to the medullary nucleus raphe magnus. The present study used electron microscopic immunocytochemical techniques to examine the morphology and synaptic relationships of GABA-immunoreactive terminals in the ventrolateral periaqueductal gray. These putative GABAergic terminals comprise almost 40% of all axon terminals in the periaqueductal gray. GABA-immunoreactive terminals contain small, clear, pleomorphic or round, vesicles, and 46% also contain some dense-cored vesicles. In some experiments we also used a colloidal gold-conjugated retrograde tracer to label periaqueductal gray neurons that project to the nucleus raphe magnus. About half of the synaptic inputs onto the cell bodies and proximal dendrites of retrogradely labeled neurons are GABA-immunoreactive; these putative GABAergic synapses, which directly control activity in neurons projecting from the periaqueductal gray to the nucleus raphe magnus, might mediate the antinociception-related effects of exogenous GABAA receptor ligands.     

Distribution and targets of the relaxin-3 innervation of the septal area in the rat

Neural tracing studies have revealed that the rat medial and lateral septum are targeted by ascending projections from the nucleus incertus, a population of tegmental GABA neurons. These neurons express the relaxin-family peptide, relaxin-3, and pharmacological modulation of relaxin-3 receptors in medial septum alters hippocampal theta rhythm and spatial memory. In an effort to better understand the basis of these interactions, we have characterized the distribution of relaxin-3 fibers/terminals in relation to different septal neuron populations identified using established protein markers. Dense relaxin-3 fiber plexuses were observed in regions of medial septum containing hippocampal-projecting choline acetyltransferase (ChAT)-, neuronal nitric oxide synthase (nNOS)-, and parvalbumin (PV)-positive neurons. In lateral septum (LS), relaxin-3 fibers were concentrated in the ventrolateral nucleus of rostral LS and the ventral nucleus of caudal LS, with sparse labeling in the dorsolateral and medial nuclei of rostral LS, dorsal nucleus of caudal LS, and ventral portion nuclei. Relaxin-3 fibers were also observed in the septofimbrial and triangular septal nuclei. In the medial septum, we observed relaxin-3-immunoreactive contacts with ChAT-, PV-, and glutamate decarboxylase-67-positive neurons that projected to hippocampus, and contacts between relaxin-3 terminals and calbindin- and calretinin-positive neurons. Relaxin-3 colocalized with synaptophysin in nerve terminals in all septal areas, and ultrastructural analysis revealed these terminals were symmetrical and contacted spines, somata, dendritic shafts, and occasionally other axonal terminals. These data predict that this GABA/peptidergic projection modulates septohippocampal activity and hippocampal theta rhythm related to exploratory navigation, defensive and ingestive behaviors, and responses to neurogenic stressors.     

Ontogeny of gamma-aminobutyric acid-immunoreactive neuronal populations in the forebrain and midbrain of the sea lamprey

Although brain organization in lampreys is of great interest for understanding evolution in vertebrates, knowledge of early development is very scarce. Here, the development of the forebrain and midbrain gamma-aminobutyric acid (GABA)-ergic systems was studied in embryos, prolarvae, and small larvae of the sea lamprey using an anti-GABA antibody. Ancillary immunochemical markers, such as proliferating cell nuclear antigen (PCNA), calretinin, and serotonin, as well as general staining methods and semithin sections were used to characterize the territories containing GABA-immunoreactive (GABAir) neurons. Differentiation of GABAir neurons in the diencephalon begins in late embryos, whereas differentiation in the telencephalon and midbrain was delayed to posthatching stages. In lamprey prolarvae, the GABAir populations appear either as compact GABAir cell groups or as neurons interspersed among GABA-negative cells. In the telencephalon of prolarvae, a band of cerebrospinal fluid-contacting (CSF-c) GABAir neurons (septum) was separated from the major GABAir telencephalic band, the striatum (ganglionic eminence) primordium. The striatal primordium appears to give rise to most GABAir neurons observed in the olfactory bulb and striatum of early larval stages. GABAir populations in the dorsal telencephalon appear later, in 15-30-mm-long larvae. In the diencephalon, GABAir neurons appear in embryos, and the larval pattern of GABAir populations is recognizable in prolarvae. A small GABAir cluster consisting mainly of CSF-c neurons was observed in the caudal preoptic area, and a wide band of scattered CSF-c GABAir neurons extended from the preoptic region to the caudal infundibular recess. A mammillary GABAir population was also distinguished. Two compact GABAir clusters, one consisting of CSF-c neurons, were observed in the rostral (ventral) thalamus. In the caudal (dorsal) thalamus, a long band extended throughout the ventral tier. The nucleus of the medial longitudinal fascicle contained an early-appearing GABAir population. The paracommissural pretectum of prolarvae and larvae contained a large group of non-CSF-c GABAir neurons, although it was less compact than those of the thalamus, and a further group was found in the dorsal pretectum. In the midbrain of larvae, several groups of GABAir neurons were observed in the dorsal and ventral tegmentum and in the torus semicircularis. The development of GABAergic populations in the lamprey forebrain was similar to that observed in teleosts and in mouse, suggesting that GABA is a very useful marker for understanding evolution of forebrain regions. The possible relation between early GABAergic cell groups and the regions of the prosomeric map of the lamprey forebrain (Pombal and Puelles [ 1999] J. Comp. Neurol. 414:391-422) is discussed in view of these results and information obtained with ancillary markers.     

Localization and source of gamma aminobutyric acid immunoreactivity in the isthmic nucleus of the frog Rana esculenta

The distribution of gamma-aminobutyric acid (GABA)-containing neurons and nerve fibers was studied in the isthmic nucleus of the frog Rana esculenta using light and electron microscopical immunohistochemical techniques. Approximately 0.5% of isthmic cells showed GABA immunopositivity, and the majority of these cells was found in the anterior one-third of the nucleus. A meshwork of GABA-immunostained fine beaded axons filled the entire isthmic nucleus. The GABA-immunoreactive terminals formed pericellular basket-like structures around a few cells both in the medulla and the cortex of the isthmic nucleus. To determine the source of GABA-positive fibers in the isthmic nucleus lesion experiments were carried out. After unilateral tectal ablation no change was observed in GABA immunoreactivity. Hemisectioning the tegmentum close to the anterior border of the isthmic nucleus, transection of the caudal tectal commissure and decussatio veli, or electrical lesioning of the anterodorsal tegmental nucleus all resulted in a moderate decrease in the density of GABA-positive fibers. Our results suggest that the majority of GABA-positive fibers derives from local GABA-positive cells, but some GABAergic afferents seem to arise in the tegmentum.     

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.     

Inhibitory synaptic input to identified rubrospinal neurons in Macaca fascicularis: an electron microscopic study using a combined immuno-GABA-gold technique and the retrograde transport of WGA-HRP.

Rubrospinal neurons of the magnocellular division of the red nucleus of Macaca fascicularis were retrogradely labeled following spinal cord microinjections of wheat germ agglutinin-horseradish peroxidase, as demonstrated by the chromagen tetramethylbenzidine, identifying the mesencephalic cells of origin of this descending motor pathway. The tissue was processed for electron microscopy and subsequently tested on the electron microscope grid for immunoreactivity of gamma aminobutyric acid (GABA) in presumed local circuit neuronal somata, in dendrites, and in axonal terminals. Results demonstrate the presence of retrogradely labeled rubrospinal neurons of medium and large diameters (30-90 microns) and immunoreactive neurons of small size (less than 20 microns in diameter) within the nucleus. In addition, there are substantial numbers of GABAergic, presumably inhibitory, synaptic structures contacting somata and primary, medium, and small sized dendrites, as well as spineheads of rubrospinal neurons. The immunoreactive presynaptic profiles exhibit two different morphological appearances: one axonal and the other dendritic. Axonal terminals contain densely packed pleomorphic to flattened vesicles and form primarily symmetrical synapses with somata and all regions of the dendritic arbor. GABAergic profiles resembling presynaptic dendrites (PSDs) are also present. These profiles possess scattered flattened to pleomorphic synaptic vesicles in a translucent cytoplasm and are often postsynaptic to axonal terminals of unknown origin, or to GABAergic profiles. GABAergic local circuit neurons (LCNs), the neurites of which remain within the confines of the nucleus, appear to be contacted primarily by cortical and cerebellar afferents. These LCNs may or may not possess axons and thus may represent both the source of the GABAergic axonal terminals as well as that of the PSDs. Inhibitory afferents from other sources, such as the mesencephalic reticular formation, may also account for GABAergic terminals involved in this inhibition. We propose that the level of excitability of rubrospinal neurons and their subsequent activation of spinal motor neurons and interneurons is significantly regulated by the local circuit GABAergic inhibitory interneuronal population of the nucleus proper and probably by axons entering the nucleus from an extranuclear source.     

GABA-accumulating neurons in the nucleus raphe dorsalis and periaqueductal gray in the rat: A biochemical and radioautographic study

The possibility of a GABAergic innervation of the nucleus raphe dorsalis (NRD) has been investigated by using the following approaches: (i) the identification of the principal neuronal groups afferent to the NDR by using horseradish peroxidase retrograde transport, (ii) the determination of glutamate decarboxylase activity (GAD) in the NRD after lesioning these groups or their putative pathways, and (iii) the radioautographic identification of terminals axons and nerve cells accumulating intraventricularly injected [3H]GABA. The hypothesis of a local GABAergic network is supported by the failure to obtain important changes in GAD after lesions of NRD afferents and the presence in this nucleus of terminals, fibers and nerve cell bodies accumulating [3H]GABA. It appears that these GABA-accumulating neurons could represent a portion of aperiventricular GABAergic system in the periaqueductal gray and the pontine ventricular gray.     

GABAergic neurons in the primate basal forebrain magnocellular complex

Hybridization histochemistry was used to detect messenger ribonucleic acid (mRNA) coding for glutamic acid decarboxylase, the synthesizing enzyme for gamma-aminobutyric acid (GABA), in neurons of the nucleus basalis of Meynert and nucleus of the diagonal band of Broca of one rhesus monkey and 4 baboons. GABAergic neurons were distributed among the unlabeled large, hyperchromic Nissl-stained neurons characteristic of this basal forebrain magnocellular complex, although they were infrequent within the dense islands of large cells. Most GABAergic cells were small to medium in size, but some were large and hyperchromic. These findings demonstrate a heterogeneous population of presumably inhibitory neurons in the basal forebrain magnocellular complex of primates.