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G Levi

Publications and source records attributed to G Levi.

At least 163 records · Page 9Linked to original sources
PubMed1987

Glial conditioned media inhibit the proliferation of cultured rat cerebellar astrocytes.

Conditioned medium (CM) obtained from rat cerebellar astrocytes cultured in a serum-containing medium was able to inhibit [3H]thymidine incorporation into proliferating astrocytes, when compared to fresh medium. This effect could be attributed to two fractions of the CM with different molecular weights. The low molecular weight fraction (Mr less than 1,000) inhibited the cellular transport of the labeled precursor, without significantly affecting cell proliferation. The high molecular weight fraction (Mr greater than 10,000) showed a strong inhibitory effect on astrocyte proliferation, which was documented using different assay techniques: i) [3H]thymidine incorporation performed in conditions preventing the effects of CM on transport; ii) [3H]thymidine autoradiography; iii) determination of the DNA content of the cultures. The inhibitory activity was present in media conditioned by non proliferating astrocytes treated with the antimitotic cytosine arabinoside, but not in media conditioned by neuron-enriched cultures nor in a chemically defined (N2) CM. The antiproliferative activity of astrocyte CM could be due either to a rapid depletion of mitogenic factors present in serum, or, to a secretion of growth inhibitory factor(s) by cultured astrocytes.

Animals↗
PubMed1987

The proteoglycan chondroitin sulfate is present in a subpopulation of cultured astrocytes and in their precursors.

We have used an antibody raised against the bovine nasal cartilage proteoglycan chondroitin sulfate (CS) digested with chondroitinase ABC (anti-CS serum) to stain cerebellar glial cells maintained in culture. In cultures grown in the presence of serum, the antibody stained a subclass of GFAP+ astrocytes which we have previously shown to selectively bind the monoclonal antibodies A2B5 and LB1. Also the direct bipotential precursors of these cells, capable of differentiating into GFAP+ astrocytes or into Gal-C+, O1+ oligodendrocytes depending on the culture conditions, were stained, but stopped to produce CS when they differentiated into oligodendrocytes.

Animals↗
PubMed1987

Mitogenic effect of a human placental factor on astrocytes and glial precursors.

We have characterized and partially purified a new 'factor' present in human placenta which strongly stimulates the in vitro proliferation of two immunocytochemically characterized subtypes of astrocytes and of bipotential precursors of putative fibrous astrocytes and oligodendrocytes. This 'factor' has an apparent Mr of 60-80 kD and exhibits physicochemical and chromatographic properties characteristic of polypeptides. Our observations suggest that placenta-derived growth factors (PDMF) control the proliferation of glial cells and glial precursors during fetal development.

Animals↗
PubMed1987

Extensive amino acid sequence homologies between animal lectins.

We have established the amino acid sequence of the beta-D-galactoside binding lectin from the electric eel and the sequences of several peptides from a similar lectin isolated from human placenta. These sequences were compared with the published sequences of peptides derived from the beta-D-galactoside binding lectin from human lung and with sequences deduced from cDNAs assigned to the beta-D-galactoside binding lectins from chicken embryo skin and human hepatomas. Significant homologies were observed. One of the highly conserved regions that contains a tryptophan residue and two glutamic acid residues is probably part of the beta-D-galactoside binding site, which, on the basis of spectroscopic studies of the electric eel lectin, is expected to contain such residues. The similarity of the hydropathy profiles and the predicted secondary structure of the lectins from chicken skin and electric eel, in spite of differences in their amino acid sequences, strongly suggests that these proteins have maintained structural homologies during evolution and together with the other beta-D-galactoside binding lectins were derived from a common ancestor gene.

Amino Acid Sequence↗
PubMed1987

Expression sequences and distribution of two primary cell adhesion molecules during embryonic development of Xenopus laevis.

Studies of chicken embryos have demonstrated that cell adhesion molecules are important in embryonic induction and are expressed in defined sequences during embryogenesis and histogenesis. To extend these observations and to provide comparable evidence for heterochronic changes in such sequences during evolution, the local distributions of the neural cell adhesion molecule (N-CAM) and of the liver cell adhesion molecule (L-CAM) were examined in Xenopus laevis embryos by immunohistochemical and biochemical techniques. Because of the technical difficulties presented by the existence of multiple polypeptide forms of CAMs and by autofluorescence of yolk-containing cells, special care was taken in choosing and characterizing antibodies, fluorophores, and embedding procedures. Both N-CAM and L-CAM were found at low levels in pregastrulation embryos. During gastrulation, N-CAM levels increased in the presumptive neural epithelium and decreased in the endoderm, but L-CAM continued to be expressed in all cells including endodermal cells. During neurulation, the level of N-CAM expression in the neural ectoderm increased considerably, while remaining constant in non-neural ectoderm and diminishing in the somites; in the notochord, N-CAM was expressed transiently. Prevalence modulation was also seen at all sites of secondary induction: both CAMs increased in the sensory layer of the ectoderm during condensation of the placodes. During organogenesis, the expression of L-CAM gradually diminished in the nervous system while N-CAM expression remained high. In all other organs examined, the amount of one or the other CAM decreased, so that by stage 50 these two molecules were expressed in non-overlapping territories. Embryonic and adult tissues were compared to search for concordance of CAM expression at later stages. With few exceptions, the tissue distributions of N-CAM and L-CAM were similar in the frog and in the chicken from early times of development. In contrast to previous observations in the chicken and in the mouse, N-CAM expression was found to be high in the adult liver of Xenopus, whereas L-CAM expression was low. In the adult brain, N-CAM was expressed as three components of apparent molecular mass 180, 140, and 120 kD, respectively; in earlier stages of development only the 140-kD component could be detected. In the liver, a single N-CAM band appears at 160 kD, raising the possibility that this band represents an unusual N-CAM polypeptide. L-CAM appeared at all stages as a 124-kD molecule.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1987

Glutamate receptor subtypes in cultured cerebellar neurons: modulation of glutamate and gamma-aminobutyric acid release.

Using cerebellar, neuron-enriched primary cultures, we have studied the glutamate receptor subtypes coupled to neurotransmitter amino acid release. Acute exposure of the cultures to micromolar concentrations of kainate and quisqualate stimulated D-[3H]aspartate release, whereas N-methyl-D-aspartate, as well as dihydrokainic acid, were ineffective. The effect of kainic acid was concentration dependent in the concentration range of 20-100 microM. Quisqualic acid was effective at lower concentrations, with maximal releasing activity at about 50 microM. Kainate and dihydrokainate (20-100 microM) inhibited the initial rate of D-[3H]aspartate uptake into cultured granule cells, whereas quisqualate and N-methyl-DL-aspartate were ineffective. D-[3H]Aspartate uptake into confluent cerebellar astrocyte cultures was not affected by kainic acid. The stimulatory effect of kainic acid on D-[3H]aspartate release was Na+ independent, and partly Ca2+ dependent; the effect of quisqualate was Na+ and Ca2+ independent. Kynurenic acid (50-200 microM) and, to a lesser extent, 2,3-cis-piperidine dicarboxylic acid (100-200 microM) antagonized the stimulatory effect of kainate but not that of quisqualate. Kainic and quisqualic acid (20-100 microM) also stimulated gamma-[3H]-aminobutyric acid release from cerebellar cultures, and kynurenic acid antagonized the effect of kainate but not that of quisqualate. In conclusion, kainic acid and quisqualic acid appear to activate two different excitatory amino acid receptor subtypes, both coupled to neurotransmitter amino acid release. Moreover, kainate inhibits D-[3H]aspartate neuronal uptake by interfering with the acidic amino acid high-affinity transport system.

Animals↗
PubMed1986

Kainic acid stimulates GABA release from a subpopulation of cerebellar astrocytes.

The effect of acute exposure to micromolar concentrations of kainic acid was studied in rat cerebellar astroglial cultures consisting of two antigenically and functionally distinct subpopulations of astrocytes. Kainate (20-100 microM) released [3H]GABA preaccumulated by A2B5-positive stellate astrocytes in a concentration-, Na+- and Ca2+-dependent way, and did not interfere with [3H]GABA transport. We hypothesize that kainate depolarizes this class of cells by activating cationic channels and that this results in an imbalance of GABAergic transmission.

Animals↗
PubMed1986

Developmental features of rat cerebellar neural cells cultured in a chemically defined medium.

We studied some aspects of the differentiation of rat cerebellar neural cells obtained from 8-day postnatal animals and cultured in a serum-free, chemically defined medium (CDM). The ability of the cells to take up radioactive transmitter amino acids was analyzed autoradiographically. The L-glutamate analogue 3H-D-aspartate was taken up by astroglial cells, but not by granule neurons, even in late cultures (20 days in vitro). This is in agreement with the lack of depolarization-induced release of 3H-D-aspartate previously observed in this type of culture. In contrast, 3H-(GABA) was scarcely accumulated by glial-fibrillary-acidic-protein (GFAP)-positive astrocytes, but taken up by glutamate-decarboxylase-positive inhibitory interneurons and was released in a Ca2+-dependent way upon depolarization: 3H-GABA evoked release progressively increased with time in culture. Interestingly, the expression of the vesicle-associated protein synapsin I was much reduced in granule cells cultured in CDM as compared to those maintained in the presence of serum. These data would indicate that in CDM the differentiation of granule neurons is not complete, while that of GABAergic neurons is not greatly affected. Whether the diminished differentiation of granule cells must be attributed only to serum deprivation or also to other differences in the composition of the culture medium remains to be established. 3H-GABA was avidly taken up also by a population of cells which were not recognized by antibodies raised against GFAP, glutamate decarboxylase, and microtubule-associated protein 2. These cells exhibited a stellate morphology, were stained by the monoclonal antibody A2B5, and have been characterized elsewhere [Levi et al, 1986] as bipotential precursors of oligodendrocytes and of a subpopulation of astrocytes bearing a stellate shape and capable of high-affinity 3H-GABA uptake.

Animals↗
PubMed1986

A reinterpretation of tyramine sympathomimetic effect and tachyphylaxis.

In this commentary, the indirect sympathomimetic effect of tyramine and the phenomenon of tyramine tachyphylaxis are reinterpreted in terms of carrier-mediated exchange processes. Extracellular tyramine would exchange with intraterminal noradrenaline and, upon repeated tyramine administration, with a mixture of noradrenaline and tyramine progressively more enriched in the pharmacologically inactive amine.

Adrenergic Fibers↗
PubMed1986

Subpopulations of rat cerebellar astrocytes in primary culture: morphology, cell surface antigens and [3H]GABA transport.

Glial fibrillary acidic protein (GFAP)-positive astrocytes in preconfluent cultures derived from postnatal rat cerebellum have been previously shown to display two distinct morphologies, one stellate and the other irregularly epithelioid. The immunofluorescence studies described here showed that these cells also possess unique surface characteristics. In cultures derived from 8-day-old animals stellate cells bound the monoclonal antibody A2B5 whereas the epithelioid cells bound another monoclonal antibody against rat neural antigen-2 (RAN2). Some stellate cells derived from 2-day-old animals also bound tetanus toxin. The A2B5 labelling of the stellate cells made it possible to follow their fate in vitro. In confirmation of previous time-lapse studies, they underwent a shape transformation as confluence was approached, ultimately attaining a form resembling that of the epithelioid cells. Autoradiographic transport studies using two tritiated gamma-aminobutyric acid (GABA) analogues cis-1,3-aminocyclohexane carboxylic acid (ACHC) and beta-alanine revealed further differences between the two types of astrocytes. Whereas [3H]ACHC was taken up solely by the stellate cells [3H]beta-alanine was transported by both cell types. In other experiments in which various inhibitors of [3H]GABA transport were used ACHC virtually eliminated uptake into the stellate astrocyte, but had little effect on the epithelioid ones. The 'neuron-like' [3H]GABA transport process in the stellate astrocytes was confirmed in experiments comparing the effect of another compound which has been proposed as an astrocyte-selective GABA transport inhibitor, 4,5,6,7-tetrahydroisoxazolo-(4,5-C)pyridin-3-ol (THPO). No discrimination was found in its effect on the uptake of [3H]GABA into either neurons or stellate astrocytes. Further autoradiographic studies following the uptake of [3H]GABA by postnatal cerebellar slices showed that astrocytes in all layers of the cerebellar cortex and white matter transported [3H]GABA in contrast to the situation in culture where the amino acid is taken up predominantly by the stellate astrocytes. The possibility is discussed that the stellate astrocytes represent a population of cerebellar fibrous astrocytes whereas the identity of the epithelioid astrocytes is less certain.

Amino Acids↗
PubMed1986

Bipotential precursors of putative fibrous astrocytes and oligodendrocytes in rat cerebellar cultures express distinct surface features and "neuron-like" gamma-aminobutyric acid transport.

When postnatal rat cerebellar cells were cultured in a chemically defined, serum-free medium, the only type of astrocyte (defined by the expression of the glial fibrillary acidic protein, GFAP) present was unable to accumulate gamma-[3H]aminobutyric acid (GABA), did not express surface antigens recognized by two monoclonal antibodies, A2B5 and LB1, and showed minimal proliferation. In these cultures, nonneuronal A2B5+, LB1+ stellate cells exhibiting "neuron-like" [3H]GABA uptake formed cell colonies of increasing size and were GFAP-. After about one week of culturing, the A2B5+, LB1+, GABA-uptake positive cell groups became galactocerebroside (GalCer) positive. Immunocytolysis of the A2B5+ cells at 3 and 4 days in vitro prevented the appearance of the A2B5+, LB1+, GABA-uptake positive cell colonies, and also of the GalCer+ cell groups. If 10% (vol/vol) fetal calf serum was added to 6-day cultures, the A2B5+, LB1+, GABA-uptake positive cell groups expressed GFAP and not GalCer. If the serum was added to the cultures 2 days after lysing the A2B5+ cells, only A2B5-, LB1-, GABA-uptake negative astrocytes proliferated. It is concluded that the putative fibrous astrocytes previously described in serum-containing cultures (which had a stellate shape and were A2B5+, LB1+, GABA-uptake positive) derive from bipotential precursors that differentiate into oligodendrocytes (GalCer+) in serum-free medium or into astrocytes (GFAP+) in the presence of serum, while the epithelioid A2B5-, LB1-, GABA-uptake negative astrocytes originate from a different precursor not yet identified.

Animals↗
PubMed1986

Altered expression of neuronal cell adhesion molecules induced by nerve injury and repair.

Peripheral nerve injury results in short-term and long-term changes in both neurons and glia. In the present study, immunohistological and immunoblot analyses were used to examine the expression of the neural cell adhesion molecule (N-CAM) and the neuron-glia cell adhesion molecule (Ng-CAM) within different parts of a functionally linked neuromuscular system extending from skeletal muscle to the spinal cord after peripheral nerve injury. Histological samples were taken from 3 to 150 d after crushing or transecting the sciatic nerve in adult chickens and mice. In unperturbed tissues, both N-CAM and Ng-CAM were found on nonmyelinated axons, and to a lesser extent on Schwann cells and myelinated axons. Only N-CAM was found on muscles. After denervation, the following changes were observed: The amount of N-CAM in muscle fibers increased transiently on the surface and in the cytoplasm, and in interstitial spaces between fibers. Restoration of normal N-CAM levels in muscle was dependent on reinnervation; in a chronically denervated state, N-CAM levels remained high. After crushing or cutting the nerve, the amount of both CAMs increased in the area surrounding the lesion, and the predominant form of N-CAM changed from a discrete Mr 140,000 component to the polydisperse high molecular weight embryonic form. Anti-N-CAM antibodies stained neurites, Schwann cells, and the perineurium of the regenerating sciatic nerve. Anti-Ng-CAM antibodies labeled neurites, Schwann cells and the endoneurial tubes in the distal stump. Changes in CAM distribution were observed in dorsal root ganglia and in the spinal cord only after the nerve was cut. The fibers within affected dorsal root ganglia were more intensely labeled for both CAMs, and the motor neurons in the ventral horn of the spinal cord of the affected segments were stained more intensely in a ring pattern by anti-N-CAM and anti-Ng-CAM than their counterparts on the side contralateral to the lesion. Taken together with the previous studies (Rieger, F., M. Grumet, and G. M. Edelman, J. Cell Biol. 101:285-293), these data suggest that local signals between neurons and glia may regulate CAM expression in the spinal cord and nerve during regeneration, and that activity may regulate N-CAM expression in muscle. Correlations of the present observations are made here with established events of nerve degeneration and suggest a number of roles for the CAMs in regenerative events.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1986

Differential distribution of cell adhesion molecules during histogenesis of the chick nervous system.

We have compared the expression of the neural cell adhesion molecule (N-CAM) and the neuron-glial cell adhesion molecule (Ng-CAM) during histogenesis of the chick nervous system. Data from immunohistochemistry and photometry were combined to construct maps of the overall distribution and dynamics of CAM appearance and disappearance. Each CAM appeared in a characteristic spatial and temporal pattern in various areas during cell movement, fiber outgrowth, tract formation, and myelination. N-CAM was more uniformly distributed than Ng-CAM and was present on all neural cell bodies and processes of the CNS and PNS. In the adult, the staining pattern of N-CAM remained similar to that in the embryo, although the staining intensity was diminished. During embryonic development, Ng-CAM was expressed on extending neurites and migrating neurons. The appearance Ng-CAM in the CNS was correlated particularly with times of cell migration in spinal cord and cerebellum, and in regions undergoing neurite extension, such as the developing white matter of the spinal cord, the optic nerve, and the medial longitudinal fasciculus. Cell bodies not undergoing migration were negative for Ng-CAM. In the adult CNS, Ng-CAM was markedly decreased in myelinated fiber tracts like the white matter of the spinal cord but persisted in unmyelinated regions such as the olfactory bulb. In contrast, in the PNS (for example, the dorsal root ganglion and sciatic nerve), Ng-CAM appeared early on both cell bodies and neurites, and it continued to be present on both in the adult, even in the presence of myelin. Maps comparing the relative distribution of Ng-CAM and N-CAM showed dynamic reversals as the nervous system developed and, as a result, the pattern of CAM expression was markedly different in embryos and adults. This difference appears to reflect changes in the roles of selective adhesion and of the two neuronal CAMs at different times of development.

Animals↗
PubMed1986

Reading strategies in children with learning disabilities and minor neurological dysfunction.

The connexions between minor neurological dysfunction (MND) and learning disabilities (LD) are analysed in 48 children with LD with and without MND. The two groups did not differ in a set of reading/writing tests or in neuropsychological tests. What proved to be different were the profiles of correlation internal to reading and between reading and presupposed neuropsychological functions. The results highlight the importance of studying the connexions between MND and LD and the need to take neuromotor parameters into account in the construction of clinical subtypes of learning disabilities.

Child↗
PubMed1985

Dopamine decreases cell excitability in rat striatal neurons by pre- and postsynaptic mechanisms.

The mechanism by which dopamine (DA) decreases the amplitude of the EPSP-IPSP sequences evoked by cortical stimulation was investigated by means of electrophysiological and biochemical methods. Intracellular recordings indicate that DA decreases the amplitude of the excitatory and inhibitory events by reducing the increase in membrane conductance measured at the peaks of the EPSP-IPSP. The non-synaptic input resistance was not modified. In addition the catecholamine (+50/+200 nA balanced current) was shown to decrease the action of glutamate (-30/-80 nA balanced current) and GABA (+40/+100 nA balanced current) when iontophoretically applied. These observations suggest that DA interferes with the excitatory (glutamatergic) and inhibitory (GABAergic) transmission at the postsynaptic site in striatal neurons. However, the depression of cellular excitability elicited by DA could not be ascribed only to its interaction with synaptic transmission at the postsynaptic level. In fact the catecholamine also inhibited spike frequency driven by depolarizing pulses and decreased the depolarization-induced release of glutamate at the presynaptic site, as shown by biochemical experiments with striatal synaptosomal preparations. A neuromodulatory role of DA in the depression of the excitability of striatal neurons by presynaptic and postsynaptic mechanisms is suggested.

Animals↗
PubMed1985

Isolation and characterization of chicken thymic electrolectin.

We have detected the presence of a beta-D-galactoside-binding lectin (electrolectin) in extracts of the thymus of adult chickens. This lectin was purified by affinity chromatography on a lactosyl-Sepharose column to yield 1.4 mg of pure protein from 230 g of thymus. The chicken thymic electrolectin (CTE) has an Mr of 15 300 when analysed by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and of 30 000 when analysed by gel filtration. The amino acid composition of CTE is similar to that of other electrolectins purified from human and rat lung. CTE cross-reacts immunologically, but is not identical, with electrolectins from electric-eel electric organ and from chick-embryo pectoral muscle. CTE agglutinates chicken thymocytes but does not appear to promote their mitosis.

Amino Acids↗