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A Derouiche

Publications and source records attributed to A Derouiche.

17 recordsLinked to original sources
PubMed2000

Topical correlation of increased hippocampal glutamine synthetase immunoreactivity and glutamatergic terminal fields after entorhinal cortex lesion.

Many astrocyte functions are related to glutamatergic transmission and to other synaptic functions, such as glutamate uptake and glutamate metabolism. While many of these functions can be executed by perisynaptic astrocyte processes, it is not clear how these processes are formed. One of the factors guiding them to the synapse may be synaptically released glutamate. This would explain the topical correlation between laminated glutamatergic terminal fields and laminae most intensely labeled by a cytoplasmic astrocyte marker, anti-glutamine synthetase (GS). This hypothesis was tested by selectively increasing the glutamate content in one terminal field. The rat entorhinal cortex, the origin of the glutamatergic projection to the outer molecular layer (OML) of the hippocampal fascia dentata, was lesioned electrolytically. In line with the hypothesis, GS immunoreactivity was strongly increased in the OML at 6 and 8 days postlesion. Lesion of only the medial entorhinal cortex resulted in heavily increased GS immunoreactivity only in the central portion of the molecular layer (i.e., the corresponding terminal field). The laminae affected were always separated from neighboring fields by a straight and clear-cut line. Although many other factors are released in the terminal field after lesion, the results are consistent with a guiding role for glutamate. The lamina-specific effect suggests that the factor(s) involved have a very limited diffusion distance. The straight border line between affected and unaffected laminae, which cuts across astrocyte territories, can best be explained by ramification of only those processes of a given astrocyte that are contained within the lamina affected.

Animals↗
PubMed1999

Cortical neurons immunoreactive for the potassium channel Kv3.1b subunit are predominantly surrounded by perineuronal nets presumed as a buffering system for cations.

Perineuronal nets (PNs) are known as chondroitin sulphate-rich, lattice-like coatings of the extracellular matrix. In the cortex of mammalian species investigated so far, they were mainly found around GABAergic neurons, but to a lesser degree also around pyramidal cells. Previous investigations in the rat revealed similar distribution patterns of fast-firing neurons expressing both the Kv3.1b subunit of voltage-gated potassium channels and the calcium-binding protein parvalbumin. In the present study, triple fluorescence labelling was applied for the simultaneous demonstration of PNs with the N-acetylgalactosamine-specific Wisteria floribunda agglutinin (WFA), parvalbumin-immunoreactivity (ir) with a monoclonal antibody and of Kv3.1b-ir with several rabbit antibodies. Subsets of non-pyramidal neurons - enwrapped by PNs and expressing parvalbumin and Kv3.1b - were detected in the rat and monkey neocortex and hippocampus. In the rat, faintly stained PNs were additionally found around several layer II/III and V pyramidal cells immunonegative for Kv3.1b, but contacted by Kv3.1b-containing boutons. In the monkey, more intensely labelled PNs frequently occurred around pyramidal cells which themselves appeared to be Kv3. 1b-immunopositive. We also observed minor Kv3.1b-ir and parvalbumin-ir cortical cell populations which were devoid of PNs; occasionally, nets were detected around neurons lacking both immunoreactivities. By confocal laser scanning microscopy, Kv3.1b-ir and WFA-binding sites were found adjoining at the soma and proximal dendritic surface, while lectin-binding sites usually extended on more distal dendritic segments and the axon initial segments which failed to express detectable Kv3.1b-ir. This spatial relationship of both markers was also confirmed by combined WFA-gold labelling and Kv3.1b-immunoperoxidase staining at the electron microscopic level. The data are used for a critical examination of current hypotheses concerning the functional role of PNs. We conclude that PNs may serve as rapid local buffers of excess cation changes in the extracellular space. Somatic membranes of fast-spiking neurons seem to be a main, but not the only source of such changes.

Animals↗
PubMed1999

The dopamine D2 receptor subfamily in rat retina: ultrastructural immunogold and in situ hybridization studies.

Dopamine, a major neurotransmitter in the vertebrate retina, is released from interplexiform cells and a restricted subset of amacrine cells. Dopamine effects vary between different retinal cell types, most likely due to differences in cell-specific receptor subtype expression. Identification of cells expressing receptors of the D2-subfamily (D2R, D3R, D4R) on a light microscopical level has rendered equivocal results, and no information is as yet available concerning the subcellular distribution of receptor protein. In the present study, D2R and D2/3R subtype-specific antisera, and D2R-, D3R- and D4R-specific oligonucleotide probes were used for ultrastructural and in situ hybridization analyses of the receptor subtype distribution in the rat retina. Light and electron microscopy showed that in addition to the known localization of intense D2R-immunoreactivity in all dopaminergic cells immunoreactive for tyrosine hydroxylase (TH), homogeneous, less intense D2R-immunoreactivity was also seen throughout the inner plexiform layer (IPL). Ultrastructurally, many additional amacrine cell processes devoid of TH-immunoreactivity at all levels of the inner plexiform layer were immunoreactive. D2R-immunoreactivity was found mainly on intracellular vesicles, and immunoreactivity associated with the plasma membrane was always extrasynaptic. No D2R-immunoreactivity was found in amacrine cell somata postsynaptic to the so-called dopaminergic 'ring endings'. Many D2R-mRNA reactive cells were observed throughout the inner nuclear layer. Morphologically, labelled cells resemble amacrines and bipolars but not horizontal cells. Reactivity with splice variant-specific oligonucleotide probes suggested that the D2LR variant is the predominant if not the only D2R isoform in the rat retina. D2R-mRNA reactivity was not observed in other retinal layers, in particular not in photoreceptor inner segments, which displayed D4R-mRNA reactivity. D3R-mRNA reactivity was not detected. The results indicate that D2-like responses are mediated through the D2R subtype, by an autoreceptor mechanism in dopaminergic cells, and by volume transmission in non-dopaminergic cells of the inner retina. D2-like responses in photoreceptors probably represent D4R activation.

Animals↗
PubMed1998

Glutamine synthetase activity in patients with Parkinson's disease.

According to current concepts, the excitatory amino acid glutamate is involved in the pathogenesis of Parkinson's disease (PD). Overactivity of glutamatergic projection neurons and beneficial effect of antiglutamatergic substances in animal experiments suggest that excess supply of glutamate might contribute to the pathophysiology of PD. Reduced activity of the glutamate metabolizing enzyme glutamine synthetase (GS) leads to decreased uptake of glutamate and thus abundant glutamate. Here we report that PD patients and age-matched controls are comparable with respect to GS activity in peripheral blood mononuclear cells (PBMC). These results imply no systemic dysregulation of the enzyme GS in patients with PD.

Case-Control Studies↗
PubMed1996

Identified glial cells in the early postnatal mouse hippocampus display different types of Ca2+ currents.

Based on their typical pattern of membrane currents, four populations of glial cells could be identified in thin brain slices of the postnatal hippocampus. In the present study, we applied the patch-clamp technique to glial cells in the hippocampal CA1 region, which are characterized by a complex pattern of different Na+ and K+ currents ("complex" cells). These cells were identified as non-neuronal cells, most likely astrocytes, by their glutamine synthetase immunoreactivity. Two types of glial Ca2+ currents could be identified that differed in their kinetics and pharmacological properties. A low-voltage activated (LVA), fast inactivating component was activated at membrane potentials positive to -60 mV and reached maximum current amplitudes at about -20 mV. This current was sensitive to amiloride and thus displayed properties of neuronal LVA currents. The threshold potential of the second Ca2+ current component was at about -40 mV, and peak currents were observed at 0 mV. In contrast to the LVA component, the inactivation of these high-voltage activated (HVA) currents slowed down with increasing depolarizations. This current was sensitive to low concentrations of Cd2+ but was not affected by amiloride. A small fraction of the HVA currents was sensitive to nifedipine, and omega-conotoxin GVIA (omega-CgTx) was also found to reduce the glial HVA component. The study provides electrophysiological and pharmacological characterization of different types of Ca2+ currents in gray matter glial cells in situ.

Animals↗
PubMed1996

Possible role of the Müller cell in uptake and metabolism of glutamate in the mammalian outer retina.

It is not known how glutamate released synaptically in the outer plexiform layer of the retina is inactivated. Since there is no extracellular enzymatic degradation, glutamate released from photoreceptors is taken up intracellularly by at least one of the three transporters known, and metabolized by a glutamate degrading enzyme. In order to elucidate which of the transporters and enzymes are involved in this process, immuno-electron microscopy was carried out on retinal sections of adult albino rats, applying antiserum against either glutamine synthetase (GS) or L-glutamate-L-aspartate transporter (GLAST). Both stainings revealed immunoreactivity in Müller cells and particularly in their processes tightly ensheathing rod photoreceptor terminals. Thus, although this remains to be tested functionally, transmitter uptake and subsequent degradation at photoreceptor terminals might be preferentially controlled by GLAST and GS expressed in the fine Müller cell processes.

Amino Acid Transport System X-AG↗
PubMed1996

Spatial relationship of lectin-labelled extracellular matrix and glutamine synthetase-immunoreactive astrocytes in rat cortical forebrain regions.

Extracellular matrix proteoglycans have previously been revealed by immunocytochemical and lectin-histochemical methods as distinct perineuronal nets in the microenvironment of different types of neurons, but also as a diffuse stain throughout the neuropil in region-dependent patterns. Ultrastructural investigations of perineuronal nets in subcortical regions have demonstrated glycan components in the close vicinity of astrocyte processes, suggesting that the extracellular matrix contributes differentially to the glianeuron interface. In the present study the spatial relationship of extracellular matrix components and astrocytes was characterised at the regional and cellular level by lectin histochemistry (soybean agglutinin, Vicia villosa agglutinin, Wisteria floribunda agglutinin) and antiglutamine synthetase immunocytochemistry in the rat neocortex and hippocampus. In most cortical areas layer-specific patterns of diffuse neuropil staining revealed by the lectins could also be recognised after glutamine synthetase (GS) immunostaining. In double-labelling experiments GS-immunoreactive astrocyte processes were found to reach lectin-stained perineuronal nets. GS-immunoreactivity was often parallelled but did not coincide with the lectin label completely, but was observed to form net-like structures similar to the perineuronal lectin staining. Using immunocytochemistry with anti-GS perineuronal, net-like structures were demonstrated on certain parvalbumin-immunopositive neurons which are known to be ensheathed by lectin-stained perineuronal nets. It was evident that a single neuron may receive net-like contacts from several astrocytes and that a single astrocyte can contribute to perineuronal nets on more than one neuron. The findings support the view that N-acetylgalactosamine-containing extracellular matrix molecules and astrocytic processes are topically associated to a high degree. Different proportions of both components may specify the individual neuronal micro-environment.

Acetylgalactosamine↗
PubMed1995

Coincidence of L-glutamate/L-aspartate transporter (GLAST) and glutamine synthetase (GS) immunoreactions in retinal glia: evidence for coupling of GLAST and GS in transmitter clearance.

Our aim was to identify proteins that mediate the uptake and degradation of synaptically released glutamate, focusing on the rat retina with its well-defined glutamatergic pathways. Immunoreactivity against the L-glutamate/L-aspartate transporter (GLAST) is present in Müller cells. Ultrastructurally, even the finest glial processes, particularly those ensheathing identified structures of glutamatergic transmission (rod spherules), are immunoreactive for GLAST. Further light and electron microscopic observations revealed that also retinal astrocytes and pigment epithelial cells are immunoreactive for GLAST. No neuronal or microglial staining was observed. This is in line with uptake of exogenous [3H]glutamate previously localized specifically in Müller cells and pigment epithelium (Ehinger and Falck: Brain Res 33:157-172, 1971). Since endogenous glutamate can only be demonstrated in Müller cells if glutamine synthetase (GS) is inhibited (Pow and Robinson: Neuroscience 60:355-366, 1994), the immunocytochemical localization of GS was determined. GS immunoreactivity was found in all but only those cell types immunoreactive for GLAST. The light and electron microscopic patterns of immunoreactivity were very similar, particularly in the outer plexiform layer. The three cell types containing both GS and GLAST (Müller cells, astrocytes, and retinal pigment epithelium) are related developmentally. In the light of the two references quoted the present data indicate that the proteins mediating retinal uptake and degradation of synaptically released glutamate may be GLAST and GS, respectively, and that they may operate in concert to terminate the neurotransmitter action of glutamate.

Amino Acid Transport System X-AG↗
PubMed1994

Regeneration of axons into the trochlear rootlet after anterior medullary lesions in the rat is specific for ipsilateral IVth nerve motoneurones.

The fibre projection from the IVth nerve nucleus to the superior oblique muscle was determined quantitatively in the normal rat by defining fibre numbers in transverse sections of the IVth nerve, and neurone numbers after retrograde labelling by horseradish peroxidase (HRP) injection into the muscle. There were 183 +/- 27 (S.E.) labelled neurones in the nucleus contralateral to the injected muscle and only 2 +/- 1 ipsilateral. The ipsilateral fibre number was 234 +/- 7 and the cell/axon ratio 0.8 +/- 0.1. Extensive analysis of all HRP retrogradely labelled material revealed no central fibre contribution to the IVth nerve other than from neurones resident in the trochlear nucleus. The central portion of the trochlear nerve tract was severed at its point of decussation in the anterior medullary velum. Ninety days after lesion, 10 +/- 4 (6% of control) neurones were labelled in the ipsilateral trochlear nucleus; none were labelled in the contralateral nucleus or in any other part of the midbrain, pons, medulla, or cerebellum. The number of myelinated fibres in the IVth nerve had decreased to 21 +/- 5 (9% of control) so that the cell/axon ratio was 0.4 +/- 0.2, thus suggesting that a single motoneurone has more fibres after lesion. In electron micrographs of the IVth nerve, larger than normal numbers of unmyelinated fibres were seen. Many myelinated fibres displayed signs of abnormal myelination. After regeneration, the projection was exclusively ipsilateral and not crossed as in the normal. These findings establish that there is a high degree of specificity after regeneration since no myelinated central nervous system axons other than trochlear fibres select the IVth nerve root as a trajectory over which to regenerate.

Animals↗
PubMed1994

Glutamine synthetase immunoreactivity in the human hippocampus is lamina-specific.

Vibratome sections of immersion-fixed (4% paraformaldehyde) hippocampi from four autoptic cases without neuropathological findings were immunostained with a polyclonal anti-glutamine synthetase antibody. Immunoreactivity is restricted to astrocytes and has a layer-specific distribution: The highest levels are observed in the outer molecular layer of the fascia dentata, in the pyramidal cell layer and in the upper stratum oriens. The stratum radiatum displays particularly low immunoreaction. A conspicuous drop in immunoreactivity from the heavily stained outer molecular layer to its inner portion is marked by a clear-cut dividing line. The layer-specific staining pattern found corresponds to the laminated distribution of glutamate binding sites described by others.

Adult↗
PubMed1994

An immunocytochemical investigation of glial morphology in the Pacific hagfish: radial and astrocyte-like glia have the same phylogenetic age.

This study attempts to reconstruct the early phylogenetic history of macroglial cells among craniates. Since glia does not fossilize, such a reconstruction must be based on a cladistic comparison of glial characters in the Recent craniate taxa (hagfishes, lampreys, and gnathostomes); however, there are only few data on glial morphology and none on glial immunocytochemistry in hagfishes. Therefore, we investigated the presence and localization of various macroglia-specific epitopes in the brain and spinal cord of the Pacific hagfish, Eptatretus stouti (Myxinoidea) by means of immunocytochemistry. Antibodies directed against S100-protein and vimentin showed no cross reactivity. Antibodies directed against glial fibrillary acidic protein and glutamine synthetase labelled various glial structures. Glial fibrillary acidic protein-like immunoreactivity was observed in ependymal cells with radially oriented processes in some regions. However, throughout the entire CNS, labelling of non-ependymal cells and their processes prevailed. The processes of these cells made occasional vascular contacts and they also made contacts with neuronal perikarya. Glutamine synthetase-like immunoreactivity was also found in some processes with radial orientation and in ependymal cells; but the antibody stained mainly non-ependymal cells which gave rise to a felt-like meshwork of interdigitating fine and very fine processes penetrating the neuropil of the entire brain. Additionally, there was labelling in the walls of blood vessels and in processes enwrapping individual neurons. The occurrence of glial fibrillary acidic protein- and glutamine synthetase-like immunoreactivity in non-ependymal glial elements in the brain of hagfishes and the relative scarcity of labelling in radial glial elements necessitates a re-interpretation of the evolutionary history of glial cells. Non-ependymal macroglia with immunocytochemical and morphological characters resembling typical (mammalian) astrocytes appears to be as primitive as the various forms of radial ependymal glia.

Animals↗
PubMed1993

Loss of layer-specific astrocytic glutamine synthetase immunoreactivity in slice cultures of hippocampus.

Glutamine synthetase (GS) supposedly inactivates the excitatory neurotransmitter glutamate. By using immunocytochemistry for GS, we recently demonstrated a layer-specific, perisynaptic distribution of GS-immunoreactive astrocytes and their processes in perfusion-fixed rat hippocampi. Highest levels of immunoreactivity were found in well defined termination zones of glutamatergic hippocampal afferents. In the present study we analysed the developmental aspect of this neuron-glia interaction by using hippocampal slice cultures lacking all extrinsic afferents. Under these conditions, no layer-specific distribution of astrocytic GS immunoreactivity could be demonstrated. This suggests that the laminated distribution of GS immunoreactivity is formed in parallel with the segregated termination of hippocampal afferents. Thus, there is no predetermined pattern of GS-containing astrocytes playing a role in the segregation of extrinsic fibres. The ultrastructural localization of GS immunoreactivity in fine astrocytic processes around asymmetric, probably glutamatergic excitatory spine synapses confirms earlier in situ findings, which suggests that this arrangement is a global phenomenon of glutamatergic systems.

Animals↗
PubMed1993

Perineuronal nets provide a polyanionic, glia-associated form of microenvironment around certain neurons in many parts of the rat brain.

The nature and function of previously described perineuronal nets are still obscure. In the present study their polyanionic components were demonstrated in the rat brain using colloidal iron hydroxide (CIH) staining. In subcortical regions, such as the red nucleus, cerebellar, and vestibular nuclei, most neurons were ensheathed by CIH-binding material. In the cerebral cortex perineuronal nets were seen around numerous nonpyramidal neurons. Biotinylated hyaluronectin revealed that hyaluronan occurs in perineuronal nets. Two plant lectins [Wisteria floribunda agglutinin (WFA) and Vicia villosa agglutinin (VVA)] with affinity for N-acetylgalactosamine visualized perineuronal nets similar to those rich in anionic components. Glutamic acid decarboxylase (GAD)-immunoreactive synaptic boutons were shown to occupy numerous meshes of perineuronal VVA-positive nets. Electron microscopically, VVA binding sites were scattered throughout perisynaptic profiles, but accumulated at membranes and in the extracellular space except not in synaptic clefts. To investigate the spatial relationship between glial cell processes and perineuronal nets, two astrocytic markers (S100-protein and glutamine synthetase) were visualized at the light and electron microscopic level. Two methods to detect microglia by the use of Griffonia simplicifolia agglutinin (GSA I-B4) and the monoclonal antibody, OX-42, were also applied. Labelled structures forming perineuronal nets were observed with both astrocytic, but not with microglial, markers. It is concluded that perineuronal nets are composed of a specialized type of glia-associated extracellular matrix rich in polyanionic groups and N-acetylgalactosamine. The net-like appearance is due to perisynaptic arrangement of the astrocytic processes and these extracellular components. Similar to the ensheathment of nodes of Ranvier, perineuronal nets may provide a special ion buffering capacity required around various, perhaps highly active, types of neurons.

Acetylgalactosamine↗
PubMed1991

Astroglial processes around identified glutamatergic synapses contain glutamine synthetase: evidence for transmitter degradation.

Glutamate is the main excitatory transmitter in the cerebral cortex. The physiologically high spatial and temporal resolution in glutamatergic transmission requires effective transmitter removal. Thus, a close topochemical relation to the glutamatergic synapse is a prerequisite for an enzyme involved in glutamate transmitter degradation. Here we report that immunoreactivity against glutamine synthetase (GS), one of the glutamate metabolizing enzymes, is localized in the fine astrocytic processes associated with identified glutamatergic synapses in the rat hippocampus. We suggest that glutamate transmitter is rapidly taken up by these fine perisynaptic astrocytic processes and degraded by GS.

Animals↗