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Biomedical subjects

R Massarelli

Publications and source records attributed to R Massarelli.

At least 73 records · Page 4Linked to original sources
PubMed1983

Development of cholinergic properties in nerve cell cultures in the presence of brain extract.

Cells dissociated from cerebral hemispheres of 6-day-old chick embryos were cultured either in standard nutrient medium or in the presence of a brain extract from 8-day-old chick embryo. Morphological observations showed the development of bipolar and multipolar neurons in both culture conditions and acetylcholinesterase activity was found in all neuronal cells. Brain extract stimulated the morphological maturation of neurons, expressed by the formation of fiber bundles, fine structural maturation and development of synapses rich in clear vesicles. Furthermore, acetylcholinesterase and choline acetyltransferase activities were higher in the cultures treated with brain extract. In these cultures, the values of choline acetyltransferase activity reached a peak at 10 days and then decreased. These observations are discussed with particular reference to proliferation, maturation and degeneration of cholinergic neurons.

Acetylcholinesterase↗
PubMed1983

A study on the topological distribution of phospholipids in microsomal membranes of chick brain using phospholipase C and trinitrobenzenesulfonic acid.

The transbilayer distribution of phospholipids in chicken brain microsomal membranes has been investigated using trinitrobenzenesulfonic acid and phospholipase C from Clostridium welchii. The exposure of intact microsomes to trinitrobenzenesulfonic acid showed that the labelling of aminophospholipids followed biphasic kinetics, indicating that these membranes contain a fast- and a slow-reacting pool of aminophospholipids. Use of microsomes radioiodinated on their surface led to the conclusion that the fast-reacting pool may be located on the outer leaflet of the microsomal vesicles. It contains about 35% of the phosphatidylethanolamine, 29% of the ethanolamine plasmalogens and 18% of the phosphatidylserine. The treatment of intact microsomes with the phospholipase C Cl. welchii produced the hydrolysis of 50% of the phospholipids without any loss of their permeability properties, indicating that they are not permeable to the hydrolase. Phospholipids extracted from the microsomes were hydrolyzed rapidly by the phospholipase C with the exception of phosphatidylserine and phosphatidylinositol. In intact microsomes about 90% of phosphatidylcholine, 32% of ethanolamine phospholipids and 60% of sphingomyelin were accessible to the phospholipase. These results suggest that the phospholipids have an asymmetric distribution in chicken brain microsomes, the external leaflet containing about 75% of the choline phospholipids and 25% of the aminophospholipids, whereas an opposite distribution is observed in the inner leaflet.

Animals↗
PubMed1983

Ectoglycosyltransferase activities at the surface of cultured neurons.

Glycosyltransferase activities (ectogalactosyl, ectofucosyl and ectosialyl) were studied at the external surface of exclusively neuronal cultures. An appropriate methodology gave the possibility to eliminate sources of errors due to the hydrolysis of nucleotide sugar substrates or due to cellular uptake of free sugars. Ovomucoid and asialofetuin coupled to Sepharose and Ultrogel beads were used as exogenous substrate to circumvent possible substrates pinocytosis. Ectoglycosyltransferase activities were studied as function of protein concentration, incubation time and amount of bead coupled exogenous acceptors. The data show that these enzymes are present at the external surface of the neuronal membrane; their possible role in cell - cell interactions is suggested.

Animals↗
PubMed1983

[Existence of UDP galactose: galactosyltransferase activity at the external surface of cultured neurons].

The existence of UDP-galactose:galactosyltransferase activities at the external surface of cultured neurons from chick embryo hemispheres, was demonstrated. [14C]-galactose was transferred upon cellular substrates present on the neuronal surface and upon exogenous substrates (ovomucoïd) bound to Sepharose beads. The activity of ectogalactosyltransferase was not due to a leakage of intracellular enzymes into the incubation medium nor to an intracellular enzymatic activity utilizing [14C]-galactose obtained from the hydrolysis of UDP-[14C]-galactose. Kinetics of these ectogalactosyltransferases were analysed. The results support the hypothesis of a communication and/or interaction among the nerve cell surfaces via glycoconjugates or their biosynthetic enzymes.

Animals↗
PubMed1982

Efflux of choline from neurons and glia in culture.

The efflux of radioactive choline from exclusively neuronal or glial cell cultures was dependent upon the concentrations of choline present in the cells and in the incubation medium, suggesting the possible presence of a homoexchange phenomenon between influx and efflux. The ionic dependence of the outward movement of choline from these cells showed that is could be stimulated by high K+ concentrations and by the absence of Ca2+. In glial cells, however, the efflux of choline was increased with a much lower concentration of K+ compared to neurons. The result may suggest that during nerve stimulation the release of K+ from neurons could stimulate, from glia, the efflux of choline which would then be taken up in neurons.

Animals↗
PubMed1982

Possible role of sialocompounds in the uptake of choline into synaptosomes and nerve cell cultures.

Incubation of primary nerve cell cultures and of crude synaptosomal preparations with neuraminidase released sialic acid from both gangliosides and sialoglycoproteins. After this treatment, the pattern of ganglioside distribution was severely modified with a decrease of polysialogangliosides (GD1b, GT1b, Gt1L, GQ1) and a dramatic increase in monosialoganglioside GM1. The choline influx into neuraminidase treated cells and organelles was reduced by 30--50% but the efflux was unmodified. In particular the high affinity mechanism of choline uptake disappeared and the low affinity mechanism was modified in both cases. The disappearance of the high affinity uptake mechanism was not followed by a decreased acetylcholine synthesis as it should be if the current theories on choline uptake and acetylcholine synthesis are correct. Our present data thus confirm our previous hypothesis that choline metabolism regulates choline uptake rather than the other way round as is suggested by the theories most widely accepted at present. Choline uptake was unaffected by pretreatment of cells and organelles with tetanus toxin suggesting that the effect of neuraminidase on the choline uptake were either mediated through glycoproteins or through gangliosides other than those which bind to tetanus toxin (GD1b and GT1b). Several speculative models for explaining the effect of neuraminidase on choline uptake are proposed.

Animals↗
PubMed1981

[Role of sialo compounds in choline uptake by synaptosomes].

Two kinetic mechanisms are responsible for the uptake of choline in Rat brain synaptosomal fractions with different affinity towards the substrate (high affinity mechanism: Km 5.1 microM, Vmax 44 pmol/mg prot/min.; low affinity mechanism: 35.7 microM, Vmax 286 pmol/mg prot/min). The treatment of synaptosomal fractions with neuraminidase (15 min. at 37 degrees C) followed by 1 min. incubation with various concentrations of [14C] choline reduces the uptake in parallel with the disappearance of the high affinity mechanism (Km 202.2 microM, Vmax 119 pmol/mg prot/min.), without influencing the synthesis of acetylcholine.

Acetylcholine↗