Glutamic Acid Decarboxylase Immunoreactive Neurons Research Articles

GABA acting on GABAB receptors located in a medullary pain facilitatory area enhances nociceptive behaviors evoked by intraplantar formalin injection.

The dorsal reticular nucleus (DRt) plays a key role in facilitation of nociceptive transmission at the spinal cord. In this study, we evaluated the mechanisms involved in GABA-mediated control of the DRt focusing on the role of local GABAB receptors. First, we used in vivo microdialysis to study the release of GABA in the DRt during the course of the formalin test. An increase of GABA levels in comparison with baseline values was detected in the second phase of the test. Because we previously showed that GABAB receptors are expressed by opioidergic DRt neurons, which respond to nociceptive stimuli and inhibit spinally projecting DRt neurons involved in descending pronociception, we then interfered with local GABAB receptors using gene transfer and pharmacological approaches. Lentiviral-mediated knockdown of GABAB1a expression decreased nociceptive responses during the second phase of the test. Local administration of the GABAB receptor antagonist CGP 35348 also decreased nociceptive responses in the second phase of the test, whereas the opposite was detected after injection of the GABAB agonist baclofen. Finally, we determined the GABAergic afferents of the DRt, namely those arising from its main brain afferents, which are located at the telencephalon and diencephalon. For that purpose, we combined retrograde tract-tracing from the DRt with immunodetection of glutamate decarboxylase, the GABA-synthesizing enzyme. The higher numbers of retrogradely labelled glutamate decarboxylase-immunoreactive neurons were located at insular, somatosensory, and motor cortices. Collectively, the results suggest that GABA acting on GABAB receptors may enhance pain facilitation from the DRt during inflammatory pain.

Closing the Gate in the Limbic Striatum: Prefrontal Suppression of Hippocampal and Thalamic Inputs

Many brain circuits control behavior by integrating information arising from separate inputs onto a common target neuron. Neurons in the ventral striatum (VS) receive converging excitatory afferents from the prefrontal cortex (PFC), hippocampus (HP), and thalamus, among other structures, and the integration of these inputs is critical for goal-directed behaviors. Although HP inputs have been described as gating PFC throughput in the VS, recent data reveal that the VS desynchronizes from the HP during epochs of burst-like PFC activity related to decision making. It is therefore possible that PFC inputs locally attenuate responses to other glutamatergic inputs to the VS. Here, we found that delivering trains of stimuli to the PFC suppresses HP- and thalamus-evoked synaptic responses in the VS, in part through activation of inhibitory processes. This interaction may enable the PFC to exert influence on basal ganglia loops during decision-making instances with minimal disturbance from ongoing contextual inputs.

Increased density of GAD65/67 immunoreactive neurons in the posterior subiculum and parahippocampal gyrus in treated patients with chronic schizophrenia

Objectives. Alterations of glutamic acid decarboxylase (GAD) play a crucial role in schizophrenic pathology. While GAD has been studied in several brain regions, its expression in the posterior hippocampus formation has not been investigated in schizophrenia. Methods. We studied the brains of 17 patients with chronic schizophrenia and 15 controls. Using the optical dissector method we counted GAD65/67 immunoreactive neurons and pyramidal cells in the posterior hippocampus, subiculum, and parahippocampal gyrus, and measured the cortical thickness in posterior subiculum and parahippocampal gyrus. Patients had received typical neuroleptics for the mean of 20.8 years. Results. In the patients we observed a significant increase of GAD immunoreactive neurons in the subiculum (left/right P = 0.004) and the parahippocampal gyrus (left P = 0.001, right P = 0.006). The hippocampus showed no or only subtle trends towards higher GAD densities. The density of pyramidal neurons and cortical thickness did not differ between the groups. A significant association between GAD density and the duration of illness was found in women with schizophrenia. Conclusions. The current data on GAD65/67 indicates a dysregulation of the GABAergic system in schizophrenia patients that may be associated with cognitive decline. However, a long term effect of neuroleptics on the GABAergic system cannot be excluded.

Vagus nerve stimulation may protect GABAergic neurons following traumatic brain injury in rats: An immunocytochemical study

Seizures and subclinical seizures occur following experimental brain injury in rats and may result from inhibitory neuron loss. This study numerically compares cortical and hippocampal glutamic acid decarboxylase (GAD) positive neurons between sham fluid percussion injury (FPI), FPI with sham Vagus Nerve Simulation (VNS), and FPI with chronic intermittent VNS initiated at 24 h post FPI in rats. Rats ( n = 8/group) were prepared for immunocytochemistry of GAD at 15 days post FPI. Serial sections were collected and GAD immunoreactive neurons were counted in the hippocampal hilus and two levels of the cerebral cortex. Numbers of quantifiable GAD cells in the rostral cerebral cortices were different between groups, both ipsilateral and contralateral to the FPI. Post hoc analysis of cell counts rostral to the ipsilateral epicenter, revealed a significant 26% reduction in the number of GAD cells/unit area of cerebral cortex following FPI. In the FPI-VNS group, this percentage loss was attenuated to only an 8.5% reduction, a value not significantly different from the sham group. In the contralateral side of the rostral cerebral cortex, FPI induced a significant 24% reduction in GAD cells/unit area; whereas, the VNS-treated rats showed no appreciable diminution of GAD cells rostral to the contralateral epicenter. Hippocampal analysis revealed a similar reduction of GAD cells in the FPI group; however, unlike the cortex this was not statistically significant. In the FPI-VNS group, a trend towards increased numbers of hilar GAD cells was observed, even over and above that of the sham FPI group; however, this was also not statistically significant. Together, these data suggest that VNS protects cortical GAD cells from death subsequent to FPI and may increase GAD cell counts in the hippocampal hilus of the injured brain.

Enhanced Fos expression in glutamic acid decarboxylase immunoreactive neurons of the mouse periaqueductal grey during opioid withdrawal

Previous studies using c-Fos immunohistochemistry suggest that a sub-population of neurons in the midbrain periaqueductal gray region is activated during opioid withdrawal. The neurochemical identity of these cells is unknown but cellular physiological studies have implicated GABAergic neurons. The present study investigated whether GABAergic neurons are activated in the mouse periaqueductal gray during opioid withdrawal using dual-antibody immunohistochemistry for Fos and glutamic acid decarboxylase. Both chronic opioid treatment and naloxone-precipitated opioid withdrawal increased Fos expression in the periaqueductal gray, with the greatest increase being four-fold in the caudal ventrolateral subdivision following withdrawal. Neurons stained for both Fos and glutamic acid decarboxylase were greatly enhanced in all subdivisions of the periaqueductal gray following withdrawal, particularly in the lateral and ventrolateral divisions where the increase was up to 70-fold. These results suggest that activation of a subpopulation of GABAergic interneurons in the periaqueductal gray plays a role in opioid withdrawal.

Vestibulotrigeminal and vestibulospinal projections in rats: retrograde tracing coupled to glutamic acid decarboxylase immunoreactivity

Immunohistochemical experiments were performed using glutamic acid decarboxylase (GAD) to identify γ-aminobutyric acid (GABA)ergic neurons in the vestibular nuclei (VN). VN neurons projecting to the sensory trigeminal complex (STC) or to the C1–C2 segments of the spinal cord were identified by injection of wheat germ agglutinin-apo-horseradish peroxidase coupled to colloidal gold (gold-HRP), a retrogradely transported tracer, in these structures. The experiments combining injection of gold-HRP in spinal cord and GAD immunohistochemistry revealed the existence in the medial, inferior and lateral VN of GAD immunoreactive neurons projecting to the spinal C1–C2 level. Experiments combining injection of gold-HRP in the STC and GAD immunohistochemistry demonstrated that, at least, 30–50% of the vestibulo-trigeminal neurons also contained GAD. Injections of two different retrograde tracers (gold-HRP and Biotinylated dextran amine) in the STC and the spinal cord demonstrated that some VN neurons project by axon collaterals to both structures. Because of the GABAergic spinal and STC vestibular projections we assume that these VN neurons with collateral projection are GABAergic. Therefore primary afferents from the face, neck or hindlimb could be modulated by inhibitory influences from GABAergic vestibular neurons.

Immunocytochemical localization of glutamic acid decarboxylase in physiologically identified interneurons of hamster spinal laminae III–V

Neurons in Rexed’s laminae III–V of an isolated spinal cord–skin patch preparation from hamsters were recorded in whole-cell mode and stained intracellularly with biocytin. Evidence of inhibitory synaptic function was obtained via post-hoc immunofluorescent labeling with a monoclonal antibody directed against an axon terminal isoform of brain glutamic acid decarboxylase. For a subset of neurons, examination with laser scanning confocal microscopy revealed punctate accumulations of glutamic acid decarboxylase immunoreactivity within axon enlargements (1–3 μm diameter), as imaged in single optical sections and confirmed by subsequent optical scans in the orthogonal plane. Axons of glutamic acid decarboxylase-immunoreactive neurons were found to exhibit dense local terminations overlapping the soma and dendrites or bifurcated into lengthy rostrocaudal daughter branches ventral to the cell body. The degree and uniformity of immunolabeling in axonal enlargements varied considerably, even amongst boutons belonging to the same cell. Glutamic acid decarboxylase-positive neurons received input from myelinated (A) afferent fibers and responded to natural stimuli appropriate for activating responses in low threshold mechanoreceptors. These results provide evidence that two different populations of GABAergic inhibitory interneurons are involved in local and intersegmental circuits that mediate integration of mechanosensory information in the deep spinal dorsal horn.

Cyclosporin A protects striatal neurons in vitro and in vivo from 3-nitropropionic acid toxicity

The neuroprotective properties of cyclosporin A (CsA) are mediated by its ability to prevent mitochondrial permeability transition during exposure to high levels of calcium or oxidative stress. By using the mitochondrial toxin 3-nitropropionic acid (3NP), the present study assessed whether CsA could protect striatal neurons in vitro and in vivo. In vitro, 3NP produced a 20-30% reduction of striatal glutamic acid decarboxylase-immunoreactive (GAD-ir) neurons. A single treatment with CsA protected GAD-ir neurons from 3NP toxicity at lower (0.2 or 1.0 microM), but not at higher (5.0 microM) doses. Similar findings were seen when the cultures were treated twice with cyclosporin. In vivo experiments used the Lewis rat model of Huntington's disease (HD) in which a low 3NP dose was delivered subcutaneously through an osmotic minipump. Rats received unilateral or bilateral intrastriatal saline injections to disrupt the blood-brain barrier (BBB) and facilitate CsA reaching vulnerable neurons. In the first experiment, CsA treated 3NP-lesioned rats displayed significantly more dopamine-and adenosine-3;, 5;-monophosphate-regulated phosphoprotein (DARPP32-ir) neurons ipsilateral to BBB disruption compared to the contralateral intact striatum, indicating that disruption of the BBB maybe necessary for CsA's neuroprotective effects. In the second experiment, stereological counts of DARPP32-ir neurons revealed that CsA protected striatal neurons in a dose-dependent manner following bilateral disruption of the striatal BBB. Rats treated with the higher (15 or 20 mg/kg) but not lower (5 mg/kg) doses of CsA displayed greater numbers of DARRP32-ir striatal neurons relative to vehicle-treated 3NP-lesioned rats. Thus, under conditions in which CsA can gain access to striatal neurons, significant protection from 3NP toxicity is observed. Therefore, CsA or more lipophilic analogues of this compound, may be of potential therapeutic benefit by protecting vulnerable neurons from the primary pathological event observed in HD.

Intrastriatal implants of polymer encapsulated cells genetically modified to secrete human nerve growth factor: Trophic effects upon cholinergic and noncholinergic striatal neurons

Nerve growth factor selectively prevents the degeneration of cholinergic neurons following intrastriatal infusion but rescues both cholinergic and noncholinergic striatal neurons if the nerve growth factor is secreted from grafts of genetically modified fibroblasts. The present study evaluated whether grafted fibroblasts genetically modified to secrete human nerve growth factor could provide trophic influences upon intact cholinergic and noncholinergic striatal neurons. Unilateral striatal grafts of polymer-encapsulated cells genetically modified to secrete human nerve growth factor induced hypertrophy and significantly increased the optical density of choline acetyltransferase-immunoreactive striatal neurons one, two, and four weeks post-transplantation relative to rats receiving identical grafts missing only the human nerve growth factor construct. Nerve growth factor secreting grafts also induced a hypertrophy of noncholinergic neuropeptide Y-immunoreactive striatal neurons one, two, and four weeks post-transplantation. Glutamic acid decarboxylase-immunoreactive neurons were unaffected by the human nerve growth factors secreting grafts. The effects upon choline acetyltransferase-immunoreactive and neuropeptide Y-immunoreactive striatal neurons dissipated following retrieval of the implants. Immunocytochemistry for nerve growth factor revealed intense graft-derived immunoreactivity for up to 1000 microns from the capsule extending along the dorsoventral axis of the striatum. Nerve growth factor-immunoreactivity was also observed within a subpopulation of striatal neurons and may represent nerve growth factor consumer neurons which retrogradely transported graft-derived nerve growth factor. When explanted, grafts produced 2-4 ng human nerve growth factor/24 h over the time course of this study indicating that this level of continuous human nerve growth factor secretion was sufficient to mediate the effects presently observed.

Transforming growth factor α differentially affects GABAergic and cholinergic neurons in rat medial septal cell cultures

The effects of transforming growth factor α (TGFα) on low and high density cultures of fetal (embryonic day 17) rat medial septal cells were investigated and in some instances, compared to those of epidermal growth factor (EGF). In high density cultures, TGFα induces a significant increase in the number of astroglia and microglia. While the effects of TGFα on the astroglia are more pronounced when compared to EGF, those on the microglia are less notable. In addition, TGFα produces a time- and dose-dependent decrease in the activity of choline acetyltransferase (EC 2.3.1.6) and a proportional decrease in the number of acetylcholinesterase-positive neurons in these high density cultures. However, although both EGF and TGFα decreased choline acetyltransferase activity maximally at the same concentration (10 ng/ml), the latter was consistently more potent. TGFα does not affect cholinergic cell survival but the expression of their chemical phenotype and does so indirectly via the glial cells. On the other hand, TGFα directly induces a dose and time-dependent increase in glutamic acid decarboxylase activity in these high density cultures without affecting the number of glutamic acid decarboxylase immunoreactive neurons. In low density cultures, TGFα acts as a general neuronal survival factor, affecting both cholinergic and GABAergic neurons. Here TGFα's neurotrophic activity is more evident than its effects on their chemical phenotype. These results suggest that TGFα exerts distinct and differential effects on the biochemical expression of two neuronal populations in the developing medial septum maintained in high density culture. Finally, as TGFα acts as a general neuronal survival factor in low density cultures, cell to cell interactions appear to be important in the ultimate response of these cells to this growth factor.

Distribution of cholinergic, gabaergic and serotonergic neurons in the medial medullary reticular formation and their projections studied by cytotoxic lesions in the cat

As part of a larger study concerning the role of neurons in the medial medullary reticular formation in sleep-wake states, the distribution and projections of cholinergic, GABAergic and serotonergic neurons were studied within the lower brainstem of the cat. Cells were plotted with the aid of an image analysis system through the medullary reticular formation and raphe in adjacent sections immunostained for choline acetyltransferase, glutamic acid decarboxylase and serotonin. Immunostained fibres and varicosities were examined and quantified by microdensitometry in regions of the medulla, pons and upper spinal cord in normal and quisqualate-injected animals to assess the loss of local and distant projections following cytotoxic destruction of neurons in the medial medullary reticular formation. Choline acetyltransferase-immunoreactive neurons are unevenly and sparsely distributed, though none the less in significant numbers (estimated at ∼9080 in total), through the medial medullary reticular formation, and are present in all tegmental fields, including the gigantocellular (∼3700) and magnocellular (∼1760) rostrally and the ventral (∼3240) and paramedian (∼380) caudally, and are absent in the midline raphe. Glutamic acid decarboxylase-immunoreactive neurons are more evenly and densely distributed in large numbers (estimated at ∼18,720) through the medial medullary reticular formation, being present in the gigantocellular (∼5960), magnocellular (∼8260), ventral (∼2280) and paramedian (∼2220) tegmental fields, and are also numerous within the raphe magnus and pallidus-obscurus nuclei (∼3880). Serotonin-immunoreactive cells are sparse in the medial medullary reticular formation (estimated to total ∼1540), where they are mainly located in the magnocellular tegmental field (∼1340), and are concentrated in larger numbers within the raphe nuclei (∼8060). Cholinergic varicose fibres were moderately densely distributed through the medial medullary reticular formation, as well as through more distant lateral, rostral and caudal brainstem and upper spinal regions. After cytotoxic lesions focussed in the gigantocellular and magnocellular tegmental fields, a loss of ∼55% of the cholinergic neurons in the medial medullary reticular formation was associated with a minor decrease (∼35% in optical density measures) of local cholinergic fibres. Small and variable reductions in varicose fibres (and their optical density measures) were detected in distant structures (including the pontine lateral, gigantocellular and subcoerular tegmental fields and the caudal spinal trigeminal nucleus), that were none the less correlated with the number of intact medial medullary cholinergic cells, suggesting that these cells may project to distant brainstem targets, in addition to providing a minor proportion of the local cholinergic innervation of the medial medullary reticular formation. GABAergic terminals were highly densely distributed through the medullary reticular formation, as well as through distant brainstem and upper spinal cord regions. With destruction of$˛60% of the medial medullary GABAergic cells, there was a dramatic loss ($˛50% in optical density measures) of GABAergic terminals in the magnocellular and gigantocellular tegmental fields, that was correlated with the number of surviving GABAergic neurons in the region. No reduction of GABAergic varicosities was evident outside the zone of GABAergic cell destruction, suggesting that these cells provide primarily a local innervation to the medial medullary reticular formation and raphe nuclei. Destruction of a minority (∼30%) of the serotonergic cells that lie within the reticular formation (and variable related destruction of those within the midline raphe nuclei) was associated with reductions (≤50%) of the moderately dense local serotonergic varicose fibres and of the similarly dense distant fibres (in the medullary and pontine lateral tegmental fields, the pontine gigantocellular tegmental field, the subcoeruleus, the spinal trigeminal nucleus and ventral horn) that were correlated with the number of medullary serotonergic cells, establishing the existence of both local and distant projections of the serotonergic neurons. These results demonstrate the presence of a significant population of cholinergic neurons in the medial medullary reticular formation, which like medullary serotonergic neurons, provide a proportion of the local and distant brainstem and upper spinal cord innervation. In contrast, a large population of GABAergic neurons provides the major local GABAergic innervation to the medial medullary reticular formation and raphe, terminating around GABAergic as well as non-GABAergic cells, and may thus serve primarily to control, by disinhibition as well as inhibition, other projection neurons of the medullary reticular formation and raphe.

Morphological and GAD immunocytochemical properties of the dorsal lateral geniculate nucleus in a reptile

Morphological and glutamic acid decarboxylase (GAD) immunocytochemical properties of the dorsal part of the lateral geniculate nucleus were investigated in reptiles, Caiman crocodilus. GAD immunoreactive neurons, although sparse, were found in the dorsal part of the lateral geniculate nucleus. In addition, puncta and fibers immunoreactive for GAD were also observed in this same nucleus. These results in Caiman utilizing GAD are similar to observations on the dorsal part of the lateral geniculate nucleus of other reptiles in which antisera to gamma amino butyric acid (GABA) were used.

Luteinizing Hormone‐Releasing Hormone and Gamma‐Aminobutyric Acid Neurons in the Medial Preoptic Area are Synaptic Targets of Dopamine Axons Originating in Anterior Periventricular Areas

The aim of this study was to characterize further the transmitter content and the location of the parent cells of tyrosine hydroxylase-immunoreactive boutons terminating on luteinizing hormone-releasing hormone- and glutamic acid decarboxylase-immunoreactive neurons in the rat medial preoptic area. Electron microscopic immunostaining for luteinizing hormone-releasing hormone, tyrosine hydroxylase or glutamic acid decarboxylase was performed on desipramine-pretreated (to protect norepinephrine and epinephrine axons) rats which received a stereotaxic injection of 6-hydroxydopamine into the medial preoptic area anteroventral periventricular nucleus 48 h prior to sacrifice. This treatment induced acute degeneration of dopamine axon terminals characterized by the development of autophagous cytolysosomes, an early morphological sign of catecholamine axon degeneration. To further define the cells of origin of these dopamine boutons, the anterograde marker Phaseolus vulgaris leucoagglutinin was iontophoretically applied to the zona incerta. Six days later, rats received a 6-hydroxydopamine injection into the zona incerta or the lateral ventricle, and 48 h later, double immunostaining was performed for Phaseolus vulgaris leucoagglutinin and tyrosine hydroxylase, luteinizing hormone-releasing hormone, or glutamic acid decarboxylase on preoptic area vibratome sections. Following the 6-hydroxydopamine injection into the anteroventral periventricular nucleus, autophagous cytolysosome-containing degenerated axons were found in synaptic contact with both luteinizing hormone-releasing hormone and GABA neurons in the medial preoptic area, confirming that these are dopaminergic connections. Following the double injection treatment, 6-hydroxydopamine-induced degenerated, Phaseolus vulgaris leucoagglutinin-labeled dopamine axons originating in the zona incerta were not found to contact luteinizing hormone-releasing hormone-containing or GABA cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Activity‐dependent regulation of glutamic acid decarboxylase in the rat barrel cortex: Effects of neonatal versus adult sensory deprivation

Immunocytochemical techniques were used to study the effects of tactual deprivation on glutamic acid decarboxylase (GAD) containing neurons in rat somatosensory barrel cortex. In normal rats GAD immunoreactive neurons and puncta are present in all laminae, with dense patches of GAD immunoreactive puncta centered on the barrels in lamina IV. Trimming whiskers of adult rats leads to a reversible decrease of GAD immunoreactivity in barrels corresponding to trimmed hairs. Intensity of GAD staining also is reversibly altered in supragranular laminae of nondeprived barrel columns flanked by deprived barrels. This indicates that GAD levels in the barrel cortex ordinarily fluctuate with changes in sensory input. By contrast, animals whose whiskers are trimmed from birth have normal GAD staining in both deprived and nondeprived barrels. Moreover, if trimmed whiskers of neonatally deprived animals are allowed to grow to normal lengths and are retrimmed later in adulthood GAD staining is not affected. Thus early tactual deprivation disrupts mechanisms that permit modulation of transmitter enzyme levels in cortical neurons following changes in sensory experience.

Glutamate decarboxylase-immunoreactive neurons are preserved in human epileptic hippocampus

The present study was designed to determine whether inhibitory neurons in human epileptic hippocampus are reduced in number, which could reduce inhibition on principal cells and thereby be a basis for seizure susceptibility. We studied the distribution of GABA neurons and puncta by using glutamate decarboxylase (GAD) immunocytochemistry (ICC) together with Nissl stains. Using quantitative comparisons of GAD-immunoreactive (GAD-IR) neurons and puncta in human epileptic hippocampus and in the normal monkey hippocampus, we found that GAD-IR neurons and puncta are relatively unaffected by the hippocampal sclerosis typical of hippocampal epilepsy where 50-90% of principal (non-GAD-IR) cells are lost. GAD-IR neurons and puncta were not significantly decreased compared with normal monkey. In 6 patients, prior in vivo electrophysiology demonstrated that the anterior hippocampus generated all seizures. The anterior and posterior hippocampus were processed simultaneously, and the counts of hippocampal GAD-IR neurons were numerically greater in anterior than in the posterior hippocampus, where no seizures were initiated. These results indicate that GABA neurons are intact in sclerotic and epileptogenic hippocampus. Computerized image analysis of puncta densities in fascia dentata, Ammon's horn, and subicular complex in epileptic hippocampi (n = 7) were not different from puncta densities in the same regions in normal monkey (n = 2). Hence, despite the significant loss of principal cells (50-90% loss) GABA terminals (GAD-IR puncta) were normal, which suggests GABA hyperinnervation of the remnant pyramidal cells and/or dendrites in human epileptic hippocampus. The apparent increase in puncta ranged from 2 (fascia dentata) to 3.3 (CA1) times normal puncta densities. These findings would suggest increased inhibition and less excitability; however, those regions were epileptogenic. We suggest that GABA terminal sprouting or hyperinnervation of the few remnant projection cells may serve to synchronize their membrane potentials so that subsequent excitatory inputs will trigger a larger population of neurons for seizure onset in the hippocampus and propagation out to undamaged regions of subiculum and neocortex.

Mossy fiber synapses on glutamate decarboxylase-immunoreactive neurons: evidence for feed-forward inhibition in the CA3 region of the hippocampus.

Mossy fibers are known to form excitatory synapses on pyramidal neurons in regio inferior of the hippocampus. This study demonstrates that the mossy fibers also establish synaptic contacts with glutamate decarboxylase-immunoreactive, supposedly GABAergic inhibitory neurons in the CA3 region. The observed connection provides a morphological basis for feed-forward inhibition of the pyramidal cells.

Glutamate decarboxylase immunoreactivity in the rat interpeduncular nucleus: A light and electron microscope investigation

The distribution of immunohistochemically demonstrable glutamate decarboxylase, the synthetic enzyme for GABA, was examined in the rat interpeduncular nucleus at the light and electron microscope levels. Immunoreactive perikarya were distributed in a characteristic pattern among the subnuclear divisions. The rostral, ventral and caudal portions of the nucleus possessed numerous immunoreactive perikarya, while few immunoreactive somata were observed in the subnuclei of the dorsal aspect. A dense field of immunostained axons and terminals was also present throughout. Ultrastructural examination of glutamate decarboxylase immunoreactivity revealed numerous labelled somata, dendritic processes, axons and boutons. Axodendritic and axosomatic synapses with immunoreactive postsynaptic profiles were numerous throughout those subnuclei with large numbers of immunoreactive somata. Immunostained terminals in contact with both immunoreactive and non-immunoreactive somatic and dendritic profiles were also present. An abundance of immunostained terminals was observed in the subnuclei that possessed a sparse population of immunoreactive somata. Immunoreactive myelinated axons of unknown origin were also present. This investigation demonstrates that the rat interpeduncular nucleus possesses a large population of glutamate decarboxylase-immunoreactive neurons coextensive with a plexus of immunostained axons and terminals. The results suggest that the immunoreactive neurons give rise to axons which contribute to an intrinsic circuit interconnecting the different subnuclear divisions. These immunoreactive neurons are in receipt of non-immunoreactive afferent inputs of variable morphology, as well as projections from intrinsic immunoreactive neurons.

Anatomy of glutamic acid decarboxylase immunoreactive neurons and axons in the rat medial geniculate body.

This is a study of the form, density, and distribution of glutamic acid decarboxylase (GAD) immunoreactive neurons and puncta (axon terminals) in the adult rat medial geniculate complex. GAD-positive elements were stained by either the peroxidase-antiperoxidase or avidin-biotin procedures. Thalamic architectonic subdivisions were defined independently in Golgi, Nissl, plastic-embedded semi-thin, and fiber-stained preparations, and from investigations of medial geniculate connectivity. GAD-positive neurons represent only approximately 1% of medial geniculate neurons. They occur in the three major medial geniculate subdivisions (ventral, dorsal, and medial). There is variability between subdivisions in the form and number of such neurons, and among the puncta. In the ventral division, immunopositive somata may have sparsely branched dendrites as long as 300-400 microns and capped with varicose expansions or bouton-like sprays of appendages. These closely appose the somata or primary dendrites of other cells; the axons of these GAD-positive neurons are also immunostained. In the dorsal division there are fewer GAD-positive neurons and their structure is different. Their dendrites are rarely immunoreactive for more than 100-150 microns; nor can their immunostained axons be traced very far. In the medial division the number of GAD-positive neurons, considering the relatively small size of this division, was high. These neurons rarely have immunostained dendrites, and more than one type of neuron is immunoreactive. The average somatic diameter of GAD-positive neurons is about 60% of that of non-immunostained cells in semi-thin material; however, the range of somatic area and the dendritic variability of these neurons suggest that cells representing more than one population are immunopositive and include all but the largest neurons. The puncta also show regional differences. Small (0.5-2 microns in diameter), medium (2-3 microns), or large (greater than 3 microns) puncta occur. In the ventral division, the predominantly medium-sized puncta are about four times as numerous on a unit/area basis than in the dorsal division, where they are far smaller and more delicate; medial division puncta are as numerous as those in the ventral division, but are much larger and coarser, and may form perisomatic arrangements. Controls were devoid of specific immunostaining.(ABSTRACT TRUNCATED AT 400 WORDS)

Glutamate decarboxylase-immunoreactive neurons in the aging rat hippocampus are more resistant to ischemia than CA1 pyramidal cells

Glutamate decarboxylase (GAD)-immunoreactive, supposedly GABAergic inhibitory, neurons in various fields of the rat hippocampus and pyramidal cells in area CA1 were quantified 1 week after transient cerebral ischemia by 4-vessel occlusion. Whereas the number of CA1 pyramidal cells in Toluidine blue-stained semithin sections were found reduced by 50% when compared with controls there was no loss of GAD-immunoreactive cells in vibratome sections of hippocampus proper and fascia dentata. These data suggest that GABAergic hippocampal neurons are more resistant to ischemia than CA1 pyramidal cells.

Glutamate decarboxylase immunoreactive neurons in rat neostriatum: their morphological types and populations

Morphological types and populations of glutamate decarboxylase (GAD)-immunoreactive neurons in rat neostriatum (Str) were studied. Str of colchicine-treated animals contained 3 types of neurons immunoreactive for GAD. The first type, which makes up 80–84% of Str neurons, was medium in size and showed moderate intensity GAD-staining. The somatic morphology of the neurons was identical to the medium-spiny projection neuron. The second type, 3–5% of Str neurons, was small to medium in size and was in-intensely stained for GAD. The somata of the neurons were round or oval and contained a narrow ring of cytoplasm surrounding the nucleus, which often had nuclear invaginations. There were only a few in each section of the third type, which were large, polygonal, and intensely stained. GAD-immunoreactive neurons, including all 3 types, ranged from 85–87% of the total neuron population. The present study indicated that GABAergic neurons in the Str are not a single morphological type and that most Str projection neurons are GABAergic.