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C Léránth

Publications and source records attributed to C Léránth.

At least 19 recordsLinked to original sources

Distribution of calbindin D28k immunoreactive cells and fibers in the monkey hippocampus, subicular complex and entorhinal cortex. A light and electron microscopic study.

Calbindin D28k (CB)-containing neurons and axon terminals in the hippocampus, subicular complex and entorhinal cortex of the African green monkey (Cercopithecus aethiops) were studied by light and electron microscopic immunocytochemistry. CB was present in granule cells of the dentate gyrus, pyramidal neurons of hippocampal fields CA1 and CA2, and in pyramidal neurons of the prosubiculum and entorhinal cortex. In contrast, pyramidal neurons in the CA3, subiculum and presubiculum were not labeled. A subpopulation of non-principal neurons (non-granule and non-pyramidal cells) was also stained for CB. These cells were rare in the hippocampus and subiculum, but were more frequently observed in the presubiculum, parasubiculum, and in the entorhinal cortex. In the electron microscope, these non-principal cells displayed fine-structural characteristics of GABAergig neurons. Strongly stained CB-immunoreactive bundles of myelinated axons were found in the molecular layer of the subiculum and in various layers of the presubiculum. The CB-positive, unmyelinated axons of the granule cells, the mossy fibers, gave rise to distinct fiber bundles. Mossy fiber terminals formed asymmetric synapses on large spines in the hilus and CA3. In addition to the giant mossy fiber boutons, there were large CB-positive terminals that formed asymmetric synapses with dentritic spines throughout the hippocampal formation. These boutons also formed axo-dendritic synapses in the entorhinal cortex. Axon terminals that formed symmetric synapses and might, thus, be derived from non-principal neurons, were rarely found in the hippocampus and subicular complex. They were more frequent in the parasubiculum and entorhinal cortex. These CB-positive terminals were small, heavily immunostained, and formed symmetric axo-dendritic synapses. Our results demonstrate a great diversity of CB-containing neurons, axons, and terminals in the monkey hippocampal formation. In general, regions that received a dense innervation of CB-positive terminals displayed pyramidal neurons that all lacked this calcium-binding protein. Further studies are required to understand the functional significance of these findings.

Animals↗

The origin of somatostatin-containing nerve fibers innervating the hypothalamic supraoptic nucleus.

Light and electron microscopic studies were performed to study the connections between somatostatin (SOS)-containing nerve terminals and vasopressin (VP)-containing neurons in the rat supraoptic nucleus (SON). SOS-positive fibers innervate the SON in both the oxytocinergic and vasopressinergic areas. Using double immunostaining symmetric synaptic contacts were visualized between SOS immunoreactive boutons and the soma of VP immunopositive neurons. Surgical transection deafferentiating the SON from all possible directions do not effect the presence of SOS immunopositive fibers. These results suggest a local origin of the SOS fibers. Somatostatin-containing perikarya can indeed be found at the dorsal border of the SON at the rostral and caudal pole of the nucleus--we suggest that these cells innervate the SON. The presence of synaptic contacts between SOS fibers and VP neurons as well as the lack of these fibers in the VP deficient Brattleboro rats indicate a role for SOS in the synthesis and/or release of vasopressin in the SON.

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Cholinergic innervation of hippocampal GAD- and somatostatin-immunoreactive commissural neurons.

This study describes the cholinergic innervation of chemically defined nonpyramidal neurons in the hilar region of the rat hippocampus. Cholinergic terminals were identified by immunocytochemistry employing a monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme, and the avidin-biotin-peroxidase (ABC) technique. Nonpyramidal neurons in the hilar region were characterized by immunostaining with antibodies against glutamate decarboxylase (GAD), the gamma aminobutyric acid (GABA)-synthesizing enzyme, and somatostatin (SS). The immunoreactivity to these antibodies was detected by using biotinylated secondary antibodies and avidinated ferritin as an electron-dense marker. This electron microscopic double immunostaining procedure enabled us to demonstrate that immunoperoxidase-labeled ChAT-immunoreactive terminals established symmetric synaptic contacts on the ferritin-labeled GAD- and SS-immunoreactive hilar cells. In additional experiments at least some of the GAD- and SS-immunoreactive hilar neurons were further characterized as commissural neurons by retrograde filling with horseradish peroxidase (HRP) following an injection of the tracer into the contralateral hilus. From these triple labeling experiments, we concluded that at least some GABAergic and somatostatin-containing neurons in the hilar region, which are postsynaptic to cholinergic terminals, project to the contralateral hippocampus. Together with previous studies on the cholinergic innervation of the hippocampus and fascia dentata, our present results thus demonstrate that different types of hippocampal cells, including GABAergic and peptidergic commissural neurons in the hilar region, receive a cholinergic input.

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Corticotropin-releasing factor in the olivocerebellar tract of rats: demonstration by light- and electron-microscopic immunohistochemistry and in situ hybridization histochemistry.

The presence of corticotropin-releasing factor (CRF) in the olivocerebellar system was demonstrated in rats by light- and electron-microscopic immunohistochemistry, as well as by in situ hybridization histochemistry. CRF-like immunoreactivity was present in each portion of the olivocerebellar system: In colchicine-treated rats, CRF-immunostained cells were found in each nuclei and subunit of the inferior olive. After surgical transection of the medullary portion of the olivocerebellar tract, CRF-immunostaining accumulated in inferior olive neurons contralateral to the lesion. In situ hybridization histochemistry indicated that CRF mRNA is transcribed in inferior olive neurons for translation to CRF precursor, whereas CRF mRNA was depleted contralateral to the lesion. CRF-immunopositive fibers of the olivocerebellar tract were demonstrated ipsilateral to the transection due to an accumulation in the proximal portion of transected axons. CRF-immunopositivity was demonstrated in climbing fibers in the molecular layer of the cerebellum at both light- and electron-microscopic levels. CRF-immunostained nerve terminals (climbing fiber varicosities) established synaptic contact with dendritic spines of the Purkinje cells. These three observations provide morphological evidence suggesting that CRF might serve as a neurotransmitter in the olivocerebellar system.

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The cholinergic innervation of the rat fascia dentata: identification of target structures on granule cells by combining choline acetyltransferase immunocytochemistry and Golgi impregnation.

A monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme, was used to study cholinergic synapses on identified (Golgi stained) granule cells in the rat fascia dentata. Choline acetyltransferase immunocytochemistry was applied to 40-microns Vibratome sections cut perpendicular to the longitudinal axis of the hippocampus. Light microscopy revealed fine varicose ChAT-immunoreactive axons in all layers of the fascia dentata, i.e., in the stratum moleculare, the stratum granulosum, and the subgranular polymorph zone. Most fibers were observed in the vicinity of granule cell bodies where they ran mainly parallel to the granular layer. Next, the immunostained Vibratome sections were sandwiched between small pieces of Parafilm and piled to form a block that was covered with agar and Golgi stained. After that, the sections were separated by cutting away the agar and removing the Parafilm. Sections containing well-impregnated granule cells were gold-toned (Fairén et al., '77), embedded in Araldite, and subjected to ultrathin sectioning for electron microscopy. A total of 14 gold-toned granule cells were examined in the electron microscope for synaptic contacts with cholinergic afferents. Choline acetyltransferase-immunoreactive axon terminals were observed that established symmetric synaptic contacts with the cell bodies and dendritic shafts of the gold-toned identified granule cells. Two types of contact were observed on spines arising from gold-toned granule cell dendrites. Immunoreactive terminals established asymmetric synaptic contacts with the head of small spines and symmetric contacts with the stalk of large, complex spines. The boutons forming asymmetric synaptic contacts with the cup-shaped spine head of the complex spines were not found to be immunoreactive. Our results demonstrate that cholinergic fibers to the rat fascia dentata establish characteristic types of synaptic contact with different postsynaptic elements of granule cells, suggesting a complex function of this afferent system.

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Cholinergic neurons in the hippocampus. A combined light- and electron-microscopic immunocytochemical study in the rat.

We report here on cholinergic neurons in the rat hippocampal formation that were identified by immunocytochemistry employing a monoclonal antibody against choline acetyltransferase (ChAT), the acetylcholine-synthesizing enzyme. In general, ChAT-immunoreactive cells were rare, but were observed in all layers of the hippocampus proper and fascia dentata with a preponderance in zones adjacent to the hippocampal fissure and in the part of CA1 bordering the subiculum. All immunoreactive cells found were non-pyramidal neurons. They were relatively small with round or ovoid perikarya, which gave rise to thin spine-free dendrites. These hippocampal neurons were very similar to ChAT-immunoreactive cells in the neocortex of the same animals but were quite different from cholinergic neurons in the basal forebrain, medial septal nucleus, and neostriatum, which were larger and more intensely immunostained. Electron-microscopic analysis of ChAT-immunoreactive cells in the hippocampus and fascia dentata revealed synaptic contacts, mainly of the asymmetric type, on cell bodies and smooth proximal dendrites. The nuclei of the immunoreactive cells exhibited deep indentations, which are characteristic for non-pyramidal neurons. Our results provide evidence for an intrinsic source of the hippocampal cholinergic innervation in addition to the well-established septo-hippocampal cholinergic projection.

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The use of gold-substituted silver-intensified diaminobenzidine (DAB) and non-intensified DAB for simultaneous electron microscopic immunoperoxidase labeling of tyrosine hydroxylase and glutamic acid decarboxylase immunoreactivity in the rat medial preoptic area.

An improved gold-substituted silver intensification procedure for the peroxidase-diaminobenzidine (DAB) reaction product was developed. The method was applied in the rat medial preoptic area to label tyrosine hydroxylase (TH)-immunoreactive profiles. Following the gold toning, the same sections were immunostained for glutamic acid decarboxylase (GAD) immunoreactivity with non-intensified peroxidase-DAB. Single DAB-labeled GAD axons were found in symmetric synaptic connection with unlabeled dendrites as well as with gold-toned immunoperoxidase-containing TH neurons.

3,3'-Diaminobenzidine↗

Somatostatin immunoreactive nerve elements in the rat small intestine: light and electron microscopic studies.

Somatostatin-containing nerve elements were identified by light and electron microscopic immunocytochemistry in the rat small intestine. Labelled nerve cell bodies were found in the submucous plexus. In the myenteric plexus and the mucosa, numerous somatostatin positive fibers were found. Labelled fibers were observed in the vicinity of blood vessels, close to the smooth muscle cell membrane and to the lamina basalis of epithelial cells. In both plexuses, there were many synapses between the labelled and unlabelled nerve elements. After transection of the nn. mesenteric some of the degenerated nerve fibers originating outside the wall of the small intestine contained also somatostatin immunoreactivity. These observations suggest that somatostatin nerves not only influence the blood flow, but participate in regulation of smooth muscle activity and, by the aid of their interneuronal synapses they have a regulatory or transmitter effect on the other intrinsic nerve elements.

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Cholinergic innervation of the rat hippocampus as revealed by choline acetyltransferase immunocytochemistry: a combined light and electron microscopic study.

The cholinergic innervation of the rat hippocampus proper and fascia dentata was investigated by using a monoclonal antibody against choline acetyltransferase (ChAT). At the light microscopic level, thin varicose ChAT-immunoreactive fibers were observed mainly in the vicinity of the pyramidal and granular layers where they formed a fine network around proximal dendrites of pyramidal and granule cells. In addition, many ChAT-immuno-reactive fibers were found in the hilar region and in stratum oriens, radiatum, and lacunosum-moleculare of all hippocampal sectors. Electron microscopic analysis revealed ChAT immunoreactivity in thin unmyelinated varicose axons and terminals which established synaptic contacts. Asymmetric contacts of ChAT-immunoreactive terminals were found on small spines in the dendritic layers of the hippocampus proper and in the molecular layer of the fascia dentata. Symmetric synaptic contacts were formed on the cell bodies of pyramidal and granule cells. Both symmetric and asymmetric synaptic contacts occurred on dendritic shafts. The analysis of serial thin sections, which allows identification of postsynaptic elements, suggests that pyramidal cells, granule cells, and nonpyramidal neurons of the hippocampus receive a cholinergic input.

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Intrinsic tyrosine hydroxylase (TH) immunoreactive axons synapse with TH immunopositive neurons in the rat arcuate nucleus.

Synaptic connections were demonstrated between tyrosine hydroxylase (TH) immunoreactive axons and dendrites in the arcuate nucleus of the rat, by electron microscopic immunocytochemistry. These connections were primarily found in the lateral part of the nucleus. The results of two types of medial-dorsal-lateral surgical de-afferentation suggest that the TH-TH contacts represent intrinsic connections.

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Estrogen responsive cells in the arcuate nucleus of the rat contain glutamic acid decarboxylase (GAD): an electron microscopic immunocytochemical study.

Twenty-one days after ovariectomy (OVX) an increased frequency of lamellar cytoplasmic organelles, termed 'whorl' bodies (WB), was observed in neurons of the rat arcuate nucleus (AN). When estradiol valerate (2 mg s.c.) was injected either at the time of OVX or one week later, the frequency of WB at 21 days was reduced. The estrogen treatment resulted in a concomitant rise in the frequency of 'nematosomes' (NS), filamentous electron-dense cytoplasmic structures. In the medial part of the AN glutamic acid decarboxylase (GAD) immunopositive symmetric (Gray II) synapses were observed in contact with WB- and NS-containing cells. After colchicine treatment, GAD immunoreactivity was observed in the WB- and NS-containing perikarya in the medial AN. Some of the NS-containing cells in the lateral AN of the colchicine-pretreated animals remained immunonegative for GAD.

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Simultaneous ultrastructural demonstration of retrogradely transported horseradish peroxidase and choline acetyltransferase immunoreactivity.

In order to study the synaptic connections of neurons identified by their projection target and neurotransmitter content, we have adapted a method of combining retrograde tracing of horseradish peroxidase (HRP) and immunocytochemistry at the electron microscopic level. HRP was injected into the rat amygdala. Sections from the rostral forebrain were processed according to the 3,3'-diaminobenzidine/glucose oxidase reaction followed by choline acetyltransferase (ChAT) localization. Neurons in the ventral pallidum which contained both the diffuse immunoperoxidase reaction product (ChAT) and large electron dense bodies characteristic of retrogradely transported HRP were defined as double labeled, i.e. cholinergic neurons that project to the amygdaloid body.

Amygdala↗

Synaptic arrangements formed by serotonin-immunoreactive axons in the substantia gelatinosa of the rat spinal cord.

Ultrastructural properties of serotonin-immunoreactive axons in the rat's substantia gelatinosa were examined in order to determine the synaptic arrangements that they form in the neuropil. Some immunoreactive varicosities formed synaptic associations with somata or dendrites suggesting that serotoninergic systems are capable of influencing the activity of these structures post-synaptically. No examples of axo-axonic synapses were found. Most varicosities, however, did not form identifiable synaptic junctions, but were closely associated with somata, dendrites or axons. Some of these associations may be functional but morphological evidence for this is lacking.

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Ultrastructural evidence of amygdalofugal axons terminating on cholinergic cells of the rostral forebrain.

In the present study a double-label ultrastructural procedure was used to study amygdalofugal fibers contacting cholinergic cells of the rostral forebrain. Following horseradish peroxidase (HRP) injections into the basolateral amygdala, anterogradely transported HRP was detected in axon terminals contacting the dendrites of choline acetyltransferase-containing cells in the ventral pallidum.

Amygdala↗

Commissural afferents innervate glutamate decarboxylase immunoreactive non-pyramidal neurons in the guinea pig hippocampus.

The innervation of GABAergic hippocampal neurons by commissural fibers was investigated in the guinea pig by a combined anterograde degeneration - immunocytochemical technique. Presumed GABAergic neurons were identified by immunocytochemistry for glutamate decarboxylase (GAD) and the commissural fibers by electron-dense degeneration following contralateral transection of the fimbria. Commissural afferents were found to establish asymmetric synaptic contacts with non-pyramidal GAD-immunoreactive neurons located in subpyramidal and suprapyramidal zones of region CA1. The observed connection suggests that inhibition of pyramidal cells may occur in a feed-forward manner as postulated by electrophysiological studies.

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On the origin of the serotonergic input to the intermediate lobe of the rat pituitary.

Serotonin-containing nerve fibers have been visualized immunocytochemically in the intermediate lobe of the rat pituitary. A 50% depletion of the serotonin level in the intermediate lobe was obtained in our previous experiment in rats with pituitary stalk transection, which may represent the total neuronally derived serotonin there. In the present studies we have attempted to determine the source of these fibers by examining the effect of hypothalamic and midbrain lesions or fiber transections on serotonin levels and serotonin-containing neuronal elements. Complete hypothalamic deafferentation resulted in a significant reduction of immunostained fibers in the intermediate lobe. A 27% fall in the serotonin content (measured by HPLC and electrochemical detection) and a significant disappearance of immunostained fibers were observed after transecting the ascending fibers from the raphe nuclei towards the hypothalamus. The transection combined with the lesioning of the hypothalamic dorsomedial nuclei resulted in a 50% decrease of serotonin level in the intermediate lobe. The present data therefore suggest that serotonin fibers in the intermediate lobe may originate from cells both in the midbrain raphe and hypothalamic dorsomedial nuclei.

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Serotoninergic endings on VIP-neurons in the suprachiasmatic nucleus and on ACTH-neurons in the arcuate nucleus of the rat hypothalamus. A combination of high resolution autoradiography and electron microscopic immunocytochemistry.

A combination of immunocytochemistry with transmitter specific autoradiography at the electron microscopic level was used to identify vasoactive intestinal polypeptide (VIP)- or ACTH-containing neurons and serotoninergic nerve terminals in the suprachiasmatic (SCN) and arcuate nuclei (AN). We observed nerve terminals showing selective uptake of [3H]serotonin forming synaptic contact with perikarya or dendrites where the postsynaptic structures exhibited VIP-like immunoreactivity in the SCN and ACTH-like immunoreactivity in the AN. The findings provide ultrastructural evidence that serotoninergic nerve fibers terminate on VIP-neurons in the SCN and on ACTH-neurons in the AN.

Adrenocorticotropic Hormone↗

Depletion of substance P and somatostatin in the upper dorsal horn after blockade of axoplasmic transport.

In the upper dorsal horn of the rat lumbosacral spinal cord, substance P and somatostatin are present in two distinct and different populations of primary central afferent terminals. Substance-P-positive terminals are mainly concentrated in lamina I, while somatostatin-positive terminals are confined to lamina II. Although these two populations of primary afferent terminals differ at light- and electron-microscopic level, they are equally affected by transganglionic degenerative atrophy (TDA) which is induced by the blockade of axoplasmic transport in the segmentally related, ipsilateral sensory nerve by the local application of Vinblastin, a microtubule inhibitor. In consequence, substance P and somatostatin are depleted in the medial and intermediate portions of the upper dorsal horn, while the lateralmost area, which represents the postaxial portion of the dermatome, remains virtually intact. Substance P and somatostatin in propriospinal elements and the axonal meshwork within the dorsolateral funicle are not affected by TDA. Neurotensine, a propriospinal neuropeptide, does not show any alterations in the affected spinal segments.

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