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G Levi

Publications and source records attributed to G Levi.

At least 181 records · Page 10Linked to original sources
PubMed1985

Characterization of GABAergic neurons in cerebellar primary cultures and selective neurotoxic effects of a serum fraction.

The morphological and functional differentiation of GABAergic interneurons present in cerebellar primary cultures has been examined by means of [3H]gamma-aminobutyric acid (GABA) autoradiography and [3H]GABA depolarization-evoked release. At 2 days in vitro these neurons showed scarce accumulation of radioactivity and no Ca2+-dependent K+-evoked or veratridine-induced release of [3H]GABA. At 5 days in vitro GABAergic interneurons appeared more intensely labeled and had grown out long and often branched neuritic processes; a large Ca2+-dependent release of [3H] GABA could be evoked by high K+. At later stages the progressive increase in labeling and branching of the neuritic processes was paralleled by a further increase in the amount and Ca2+ dependence of [3H]GABA release; a tetrodotoxin-sensitive, veratridine-stimulated release was also demonstrated. The [3H]GABA-accumulating stellate astrocytes present in the culture were not responsible for the observed release of the amino acid. GABAergic neurons were also identified by indirect immunofluorescence, using antibodies to the specific marker glutamic acid decarboxylase. Total renewal of the culture medium at 7 days in vitro caused a drastic (90%) reduction in the number of GABAergic neurons and a concomitant decrease in the amount of [3H]GABA uptake and release in the cultures. The disappearance of GABAergic neurons was caused by a low molecular weight (Mr less than 1000) fraction of the serum used to supplement the basal culture medium. This serum component did not significantly influence the survival of the major neuronal population of the culture (the granule cells) and appeared to be selectively toxic for GABAergic neurons only after they had reached a quite advanced degree of morphological and functional differentiation in vitro. The toxic activity was no longer present in neuronal or glial conditioned media.

Animals↗
PubMed1984

The distribution of electrolectin in mouse: genetic and ontogenic variations.

Electrolectins are beta-D-galactoside binding lectins found in teleosts, avians and mammals. We have studied the distribution of electrolectin in several inbred strains of mice and have followed the changes in electrolectin activity occurring during development. Electrolectin activity is present in skeletal muscle, kidney, lung, heart, thymus and spleen of the young animal. The levels of electrolectin activity present in skeletal muscle, heart, lung, kidney and liver keep increasing up to 14 days after birth and then steadily decrease. In contrast, the level of electrolectin activity in the thymus and in the spleen remain constant during development. These results are discussed in view of the possible role of electrolectin in differentiation.

Aging↗
PubMed1984

Autoradiographic localization and depolarization-induced release of acidic amino acids in differentiating cerebellar granule cell cultures.

Granule cells from 8-day-old rat cerebella were grown in basal Eagle's medium with 10% fetal calf serum, for 2,5,8 or 12 days in vitro (DIV), in conditions giving a purity greater than 90%. The results obtained can be summarized as follows: (1) Light microscopic autoradiography showed that cultured granule cells and their processes can accumulate the glutamate analog [3H]D-aspartate once they have reached an advanced degree of morphological differentiation (8 and 12 DIV), but, even then, only a limited number of cells was heavily labeled. In contrast, astrocytes were heavily labeled at all stages. (2) Calcium-dependent, high [K+]-induced release, or tetrodotoxin-sensitive, veratridine-induced release of [3H]D-aspartate from granule cell-enriched cultures was detectable only in cultures of 8 or 12 DIV. (3) When subject to 3 consecutive depolarizations, cultured granule cells maintained their ability to release [3H]D-aspartate and endogenous glutamate almost unchanged. (4) Newly synthesized [3H]glutamate was autoradiographically localized in both neurons and astrocytes (the latter, however, were not preferentially labeled as with [3H]D-aspartate), but was specifically released from neuronal structures (perikarya and processes) by depolarizing stimuli.

Animals↗
PubMed1983

Prevention and therapy with electrolectin of experimental autoimmune myasthenia gravis in rabbits.

Electrolectin (EL), an endogenous beta-D-galactoside-binding lectin from Electrophorus electricus, was found to have a prophylactic and therapeutic action on the experimental autoimmune myasthenia gravis (EAMG) in rabbits. EAMG is an autoimmune disease induced by immunization with the purified acetylcholine receptor protein (AChR) and is considered to be a good model for the human disease myasthenia gravis. Simultaneous immunization with AChR and EL completely prevented the onset of myasthenic symptoms. This preventive effect was accompanied by a decrease in the recognition of AChR by anti-AChR antibodies. The administration of EL to myasthenic rabbits led, in most cases, to a complete recovery which was not accompanied by any significant change in the level of circulating anti-AChR antibodies. No evidence for an action of EL at the muscular level could be obtained. EL, however, was found to bind to rabbit lymphocytes and to stimulate their mitosis. These results suggest that EL produces its effects on EAMG by acting at the level of the immune system. It is proposed that EL may play a role in the immunological regulation of the response to self-antigen, which could be one of the biological functions of this animal lectin.

Agglutination↗
PubMed1983

Substrate specificity and developmental aspects of a presynaptic GABA receptor regulating glutamate release in the rat cerebellum.

In order to better characterize the presynaptic GABA receptors regulating glutamate release in the cerebellum [Levi and Gallo, 1981], a number of GABA agonists and GABA transport inhibitors were tested for their ability to potentiate the depolarization-induced release of the glutamate analog D-[3H]aspartate from superfused cerebellar synaptosomes. Of all the compounds tested, only those which are known to interact specifically with GABA receptors were effective when tested on synaptosomal preparations. The order of effectiveness found was the following: muscimol congruent to 3-APS greater than or equal to P4S greater than isoguvacine greater than THIP. GABA uptake inhibitors were unable to enhance D-[3H]aspartate evoked release from synaptosomes, but were effective when tested in cerebellar slices; in the latter case, the activation of the GABA receptors may be achieved indirectly, through an increase of the extracellular GABA concentrations. The substrate specificity of the presynaptic GABA receptors regulating cerebellar acidic amino acid release appears to be similar to that reported for GABA receptors in radioligand binding studies and for GABA autoreceptors. Studies on synaptosomes from immature cerebella suggested that the presence of the potentiating effect on the acidic amino acid release by GABA agonists is correlated with the development of the parallel fiber terminals, which are believed to be the main site from which glutamate is released in the adult cerebellum.

Animals↗
PubMed1983

Selective interactions of electrolectins from eel electric organ and mouse thymus with mouse immature thymocytes.

The pure electrolectin, a beta-D-galactoside binding lectin from the electric organ of the electric eel Electrophorus electricus, was found to agglutinate selectively a subpopulation of mouse thymocytes. This cell population could be separated from non-agglutinated cells by 1 g sedimentation over fetal calf serum. The agglutinated cells could be identified as immature thymocytes on the basis of the density of theta-antigen they bear on their surfaces, their mitotic activity and the absence of response to the mitogenic action of phytohemagglutinin. The immature mouse thymocytes were found to bind the endogenous mouse thymic lectin (MTL) a protein that displays the same saccharide specificity as the eel electrolectin and with which it cross-reacts immunologically. MTL is secreted by mouse thymic reticulocytes in tissue culture and its specific activity is markedly increased after depleting the thymus of its thymocytes. This finding is an indication of the possible localization of MTL in the thymic epithelium. These results are discussed in the light of our recent findings that the eel electrolectin has prophylactic and therapeutic actions on the experimental auto-immune myasthenia gravis in rabbits.

Animals↗
PubMed1983

Enrichment of differentiated, stellate astrocytes in cerebellar interneuron cultures as studied by GFAP immunofluorescence and autoradiographic uptake patterns with [3H]D-aspartate and [3H]GABA.

A study was undertaken to correlate the morphology expressed by astroglial cells in post-natal cerebellar, interneuron-enriched primary cultures, and the ability of these cells to accumulate putative neurotransmitter amino acids. Astroglial cell morphology, as studied by GFAP immunofluorescence staining showed considerable changes during the culture period considered (up to 12 days in vitro). While the total number of GFAP-positive cells decreased with time (cell multiplication was prevented by cytosine arabinoside), a progressive enrichment of stellate astrocytes (cells bearing multiple radially arranged processes) and a striking increase in size of these cells was noted. In 12 DIV cultures stellate astrocytes accounted for 70-80% of the astrocytes present, and could reach a diameter of over 300 micron. The L-glutamate analogue, [3H]D-aspartate, was avidly taken up by all the astrocytes, independently of their shape and stage of differentiation. Astroglial cell morphology as delineated by [3H]D-aspartate autoradiography was identical to that evidenced by GFAP staining. On the other hand, [3H]GABA was accumulated in substantial amounts only by the stellate astrocytes, that is by the cells showing greater morphological differentiation. Astrocytes of other shapes were only lightly labelled by [3H]GABA in 2 DIV and 5 DIV cultures, and even less at later stages. Even within the stellate astrocyte population, the extent of [3H]GABA labelling was very variable, from one cell to another. Autoradiographic examinations and the determination of the IC50s for GABA uptake inhibitors consistently indicated that the GABA transport system present in stellate astrocytes did not have the features generally attributed to a glial transport system. In fact, beta-alanine was a very weak inhibitor, while nipecotic acid and ACHC were strongly inhibitory; DABA inhibitory potency fell somewhere in between. [3H]GABA uptake into the inhibitory interneurons present in the cultures showed similar sensitivity to GABA transport inhibitors.

Animals↗
PubMed1983

Cerebellar astroglial cells in primary culture: expression of different morphological appearances and different ability to take up [3H]D-aspartate and [3H]GABA.

In non-neuronal cultures of cells dissociated from postnatal rat cerebellum astrocytes, identified by the presence of the marker protein glial fibrillary acidic protein (GFAP), displayed two distinct morphological forms. One class was stellate in shape with radially distributed fine processes, while the other class was more varied in shape being polygonal or elongate. [3H]thymidine incorporation experiments revealed that cells of both morphologies were able to incorporate this nucleoside, suggesting the capacity for both cell types to undergo cell division. An autoradiographic study of the uptake of [3H]D-aspartate and [3H]GABA revealed that whilst the two classes of astrocytes took up the aspartate to apparently the same extent, only the stellate cells were found to be heavily labeled following incubation with [3H]GABA. A study of the cultures over a 12-day period showed that there was a disappearance of the stellate astrocytes. The time of disappearance was found to be dependent upon the initial plating density; the stellate morphology was apparent longer in lower density cultures. Time lapse studies suggested that one of the reasons for the disappearance of the stellate cells might be that in fact they underwent a change in shape following certain cell-cell interactions, but cell death also has to be considered as a further possibility. The relationships between the two classes of astroglial cells in these cultures is not yet clear. The possibilities are that they represent two different types of astrocytes, or just one type at different stages of differentiation, or maybe a combination of the two possibilities.

Animals↗
PubMed1983

Abnormalities of visual evoked potentials by checkerboards in children with specific reading disability.

Visual evoked potentials by checkerboards of varying check sizes were recorded in the two hemispheres of 16 specific reading disabled and 8 normal children. In most of the disabled subjects a gross hemisphere asymmetry was assessed, while in the control group the usual evoked potential symmetry was observed. In some disabled subjects the evoked potentials had a larger amplitude in the right hemisphere, while in others the amplitude was larger in the left hemisphere. In a small subgroup the evoked potentials were symmetrical, but they had a smaller amplitude than in the control subjects. The results, giving evidence of a dysfunction in basic visual processing, are discussed in the context of current literature on clinical subgroups and the interhemispheric relationship in the dyslexic syndrome.

Child↗
PubMed1983

Muscimol potentiation of acidic amino acid release from cerebellar synaptosomes is chloride dependent.

In previous studies we have shown that the depolarization-induced release of preaccumulated acidic amino acids and newly synthesized glutamate from cerebellar synaptosomal preparations is potentiated by gamma-aminobutyric acid (GABA) agonists through a GABAergic presynaptic mechanism. Here we report a systematic analysis of the ionic requirements of the potentiating effect of muscimol on the high K+-evoked release of D-[3H]aspartate. Our studies show that: Ca2+, Na+, and Mg2+ are not required for muscimol to exert its effect; a depolarizing concentration of K+ is a necessary, but not sufficient, condition to observe the presynaptic effect in question; and a minimal Cl- concentration (50-70 mM) is also required. A possible model based on these findings is proposed.

Animals↗
PubMed1982

Antigenic cross-reactivity between electrolectins.

The antigenic cross-reactivity between purified chick, eel and mouse electrolectins (endogenous beta-D-galactoside specific lectins) have been studied using a solid phase radioimmunoassay. The immune serum raised against the eel electrolectin crossreacts both with the chick and the mouse electrolectins, while the anti-chick electrolectin anti-serum recognizes only the eel but not the mouse electrolectin. These findings are analyzed in terms of the phylogenetic distance separating the species considered; they suggest that electrolectins fulfil a fundamental biological function.

Animals↗
PubMed1982

Putative acidic amino acid transmitters in the cerebellum. I. Depolarization-induced release.

In the present investigation we studied the autoradiographic localization and the characteristics of the depolarization-induced release of acidic amino acids in in vitro rat cerebellar preparations. Light microscopy autoradiography of cerebellar slices preincubated in the presence of the non-metabolized glutamate analogue D-[3H]aspartate showed a large accumulation of radioactivity over glial cells, and very little labelling of the granule cells, whose putative neurotransmitter may be glutamate. In spite of its predominant localization in glia, D-[3H]aspartate (and [14C]glutamate) was released from cerebellar slices depolarized with high [K+] in a Ca2+-dependent way, and the release elicited by veratrine was prevented by TTX. These findings, together with the observation that freshly isolated or cultured glial cells did not show any Ca2+-dependent, depolarization-induced release of D-[3H]aspartate, suggest that the radioactive amino acid released from slices has a neuronal origin. The high [K+]-induced release of exogenous radioactive acidic amino acids from superfused cerebellar synaptosomal preparations exhibited, as best, a modest Ca2+-dependence, a result probably due to the existence of a substantial non-Ca2+-dependent release of the amino acid from glial fragments contaminating the preparation. However, both the K+-evoked release of endogenous glutamate, and that of [14C]glutamate previously synthesized from [14C]glutamine were largely Ca2+-dependent, suggesting that nerve endings are the main sites involved in the stimulus-coupled secretion. In the experiments in which synaptosomes had been prelabelled with [14C]glutamine, a study of the specific radioactivity of the glutamate released and of that present in synaptosomes at the beginning and at the end of superfusion period provided evidence in favour of a preferential release of the newly synthesized [14C]glutamate. In contrast to glutamate, endogenous aspartate was not released in a Ca2+-dependent manner, and the efflux of newly formed [14C]aspartate was only slightly potentiated by Ca2+, which suggests that glutamate and aspartate are not released from the same sites. Studies on preparations (slices and synaptosomes) from immature, 8-day-old cerebella showed that neither the K+-evoked release of D-[3H]aspartate, nor that of endogenous glutamate was Ca2+-dependent. In conclusion, the data presented are consistent with the proposition that glutamate has a neurotransmitter role in the cerebellum.U

Amino Acids↗
PubMed1982

Evidence in favor of a neurotransmitter role of glycine in the rat cerebral cortex.

In the present study we analyze whether glycine satisfies some electrophysiological and biochemical criteria to consider it as a putative transmitter in the rat cerebral cortex. Intracellular recordings from rat sensory-motor cortex showed that in 15-20% of the tested neurons glycine hyperpolarized the cell membrane, decreased the firing rate and flattened the evoked EPSP-IPSP sequence by increasing the membrane conductance. The iontophoretic application of strychnine antagonized the block of 'spontaneous' firing and the membrane hyperpolarization induced by glycine. Moreover, in a group of neurons, strychnine decreased the amplitude and duration of the IPSP and brought back the membrane potential to resting values. Previously accumulated [3H]glycine and endogenous glycine were released from cortical synaptosomal preparations by depolarizing stimuli in a Ca2+-dependent way. The release pattern of glycine was qualitatively similar in cortical and in spinal synaptosomes. [14C]Glycine was rapidly synthetized from [14C]serine in cortical synaptosomal preparations, and the newly formed [14C]glycine was released by depolarizing stimuli in a Ca2+-dependent way. It is concluded that glycine, which is generally considered as an inhibitory neurotransmitter in the spinal cord, medulla and pons, may also have a transmitter role in a discrete number of cortical neurons of some mammalian species.U

Animals↗
PubMed1982

Selective release of glutamate from cerebellar granule cells differentiating in culture.

The aim of the present study was to assess whether endogenous and newly synthesized glutamate can be released from differentiating cultured cerebellar granule cells in a way compatible with a neurotransmitter role. Granule cells from 8-day-old rat cerebella were grown in basal Eagle's medium with 10% fetal calf serum for 2-12 days in vitro (DIV), then washed with Krebs-Ringer medium, and labeled for 45 min with tracer amounts of radioactive glutamine. Subsequently, the release of endogenous glutamate and of newly formed radioactive glutamate was measured in basal conditions and upon depolarization with elevated K(+) concentration or veratridine. At 2 DIV, the release of endogenous and newly synthesized glutamate evoked by high K(+) concentration was small and Ca(2+) independent, but it progressively and steadily increased (up to 8- to 10-fold) and became Ca(2+) dependent (up to 80-85%) at later stages (4, 8, and 12 DIV). Veratridine was almost ineffective with cells at 2 DIV but greatly increased glutamate release (endogenous and neosynthesized) at 8 DIV, and its action was totally antagonized by tetrodotoxin. The level and synthesis of glutamate remained fairly constant in cells from 2 to 12 DIV. gamma-Aminobutyric acid synthesis from radioactive glutamine was about 3% of that of glutamate, and gamma-aminobutyric acid release (endogenous and neosynthesized) was not measurable. Aspartate synthesis was about 10% of that of glutamate, and the high K(+) concentration-evoked release of this amino acid was modest and scarcely affected by Ca(2+). Neither high K(+) concentration nor veratridine was able to induce glutamate release from confluent cerebellar astrocyte cultures at 14 DIV, although the level and synthesis of the amino acid were comparable to those in granule cells. In conclusion, the data show that a stimulus-coupled release of endogenous and neosynthesized glutamate is progressively expressed by cerebellar granule cells differentiating in culture, and this strongly supports the concept that glutamate is the neurotransmitter of these cells.

Animals↗
PubMed1982

Language disorders and prognosis for reading disabilities in developmental age.

2 groups of language-disordered children were studied regarding the reading levels reached at the end of the first year of school. The first group was composed by children with language retardation mainly on phonological level, the second group was composed by children with retardation also on semantic-syntactic level. The results confirmed the strong association between reading disability and language disorders and yielded significant differences between the two groups: the reading achievement seemed to be associated mainly to semantic and syntactic competences.

Achievement↗