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Biomedical subjects

B Pessac

Publications and source records attributed to B Pessac.

At least 55 records · Page 3Linked to original sources
PubMed1987

Differentiation of retrovirus-infected avian neuroretina cells.

The effects of oncogenic retroviruses on the expression of differentiation markers were studied in monolayer cultures of chick and quail embryo neuroretinas. Transformation by Rous sarcoma virus (RSV) did not affect the appearance of synapses, and the expression of glutamic acid decarboxylase was stimulated by pp60v-src, the product of the src gene. Quail embryo neuroretina cells transformed by Mill Hill 2 (which contains the two oncogenes v-mil and v-myc) were induced to proliferate into permanent cultures that synthesized crystallins and produced lentoid bodies. In contrast, transformation with a temperature-sensitive mutant of RSV reversibly blocked the production of crystallins and lentoid bodies. These data show that given cellular genes can respond differently to distinct oncogenes.

Animals↗
PubMed1987

Free amino acid content of astroglial cell clones derived from 8-day postnatal mouse cerebella.

We have measured the free amino acid content of three distinct astroglial cell clones derived from permanent lines obtained after "spontaneous immortalization" of 8-day postnatal mouse cerebellar cultures; these clones show characteristics similar to the Golgi Bergmann glia cells, the fibrous astrocytes, and the velate protoplasmic astrocytes, i.e., the three main types of cerebellar astrocytes. The relative concentrations of amino acids that are thought to act as neurotransmitters were compared in confluent cultures of the different astroglial clones. The most striking result was a high concentration of glycine (20% of free amino acids), even in astroglial cells cultured in a glycine-free medium, a finding suggesting that glycine is synthesized by the astroglial clones. Furthermore, no gamma-aminobutyric acid (GABA) was detected. In contrast, a "neuron-like" clone derived from the same cerebellar culture contained GABA, whereas its glycine content was much lower than that of the astroglial clones. The present results, together with our previous finding of glycine synthesis in an astrocytic clone derived from 14-day postnatal mouse cerebella transformed by simian virus 40, indicate that a high glycine content may be characteristic of many cerebellar astroglial types.

Amino Acids↗
PubMed1986

Crystallin gene expression and lentoid body formation in quail embryo neuroretina cultures transformed by the oncogenic retrovirus Mill Hill 2 or Rous sarcoma virus.

The lens-specific proteins alpha and delta crystallins and lentoid bodies, structures that follow a differentiation pathway similar to that of the lens, regularly appear after 4 to 5 weeks in quail embryo neuroretina monolayer cultures. We have investigated the effects of the avian oncogenic retroviruses Mill Hill 2 and Rous sarcoma virus on this process. Quail embryo neuroretina cells transformed by Mill Hill 2 virus were established into permanent cultures that synthesized alpha and delta crystallins and contained stem cells for the production of lentoid bodies. In contrast, transformation with the Rous sarcoma virus mutant tsNY-68 blocked the appearance of mRNA crystallins, but cytoplasmic alpha and delta crystallin mRNA and alpha crystallin appeared 44 h after a shift to the nonpermissive temperature. However, delta crystallins and lentoid bodies were only present after 7 days. The crystallins of transformed quail neuroretina cultures were immunologically indistinguishable from those of quail lenses and of normal quail embryo neuroretina cultures.

Animals↗
PubMed1985

Glutamic acid decarboxylase activity is stimulated in quail retina neuronal cells transformed by Rous sarcoma virus and is regulated by pp60v-src.

Rous sarcoma virus (RSV) stimulates in quail embryo neuro-retina (NR) cultures the specific activity of glutamic acid decarboxylase (GAD), the enzyme responsible for the synthesis of gamma-aminobutyric acid, a major inhibitory neurotransmitter in NR and in central nervous system. In quail embryo NR cultures transformed by ts NY-68, a thermodependent transformation-defective mutant of RSV, stimulation of GAD activity is regulated by pp60v-src, the product of the src gene of RSV. Fibroblasts and myoblasts have a very low GAD activity that is not stimulated after transformation by RSV. Neuronal clones, previously derived from ts NY-68-transformed established NR cell lines, have a high GAD activity which is regulated by pp60v-src, while other clones have a low GAD activity apparently not regulated by pp60v-src. These data indicate that pp60v-src selectively activates the expression of GAD in distinct neuronal cells of quail embryo NR cultures transformed by RSV. GAD activity is also stimulated in NR cells infected with viruses containing v-mil.

Animals↗
PubMed1985

Quail neural crest cells transformed by Rous sarcoma virus can be established into differentiating permanent cell cultures.

Quail neural crest cells derived from the truncal neural primordium, infected in vitro by Rous sarcoma virus (RSV) in January 1978, were induced to multiply and have been established into permanent cultures. These cultures contain cells that differentiate into melanocytes, neuron-like cells and flat cells. About 50% of these different cell types are tetanus-toxin positive. Electrophysiological studies have shown that some cells can generate action potentials similar to those reported in quail neural crest primary cultures. Taken together these data show that the RSV-transformed quail neural crest permanent cultures are composed of stem cells which can differentiate into cell types specific for neural crest.

Animals↗
PubMed1985

Astrocytic cerebellar cell clones synthesize the beta' isoforms of the beta-tubulin protein family.

We have analysed the isotubulin pattern of three astrocytic cell clones, derived from spontaneously established permanent cell cultures originating from 8-day postnatal mice cerebellar explants, in comparison with that of primary astroglial cultures from embryonic brain and cerebellum. These astrocytic clones, which may represent the different astroglial cell types of mouse cerebellum, did not produce the alpha- and beta-acidic isoforms, these being found only in cells of neuronal lineage. However, the three astrocytic clones, but not the primary astroglial cultures, did synthesize the beta'-tubulin isoforms; in addition quantitative analysis of the beta' proteins showed a positive correlation between the ability of the cells to extend processes and their synthesis of the beta' isoforms. These data suggest that the presence of beta'-tubulin is not specific for neuronal cells but may be related to the ability of cells from the nervous system to extend processes.

Animals↗
PubMed1984

Astrocytic cell clones derived from established cultures of 8-day postnatal mouse cerebella.

Clonal permanent cell lines with astrocytic properties have been established from explant cultures of 8-day postnatal mouse cerebella after in vitro spontaneous transformation, i.e. without the addition of carcinogens or oncogenic viruses. The cell lines were derived in a multistage process. Slowly proliferating foci with several morphologies appeared 4 months after initiation of the cultures and became progressively enriched by cells with a homogeneous appearance. These cells could be established into permanent cell lines from which many clones were obtained. Some of these cloned cell lines bound anti-GFAP sera and therefore appeared to be astrocytic. According to their morphology, 3 separate types of these GFAP-positive clones could be distinguished. Type I and II cells had small somata; type I had several short processes, while type II had two processes, one of which was very thin and long (greater than 200 microns). Type III cells had large flat somata and no processes. The three types of clonal cell lines were labeled by monoclonal antibodies which bind to astrocytes in vivo. In particular, three monoclonal antibodies (BSP-3, M2 and M3) bound only to type II cells in a distinct pattern. Type I and II astrocytes are pseudodiploid and type III, heteroploid. The properties of these different clonal cell lines are very stable. We have thus obtained permanently established clonal cultures of mouse cerebellum astrocyte-like cells, which might be the in vitro counterparts of fibrous (type I), or velamentous (type III) astrocytes and of Golgi epithelial cells (type II).

Animals↗
PubMed1983

Cells with neuronal properties in permanent cultures of quail embryo neuroretinas infected with Rous sarcoma virus.

Neuroretina cells from 7-day quail embryos infected 'in vitro' with the mutant ts NY-68 of Rous sarcoma virus, were established into permanent cultures. An initial stage of cellular proliferation was followed by a period of minimal multiplication. After recovery from this crisis, cell proliferation resumed. About 30% of the cells had binding sites for tetanus toxin and the monoclonal antibody A2B5 which seem to be specific for neurons, and an ultrastructural study suggested the presence of neurons and Müller (astroglial) cells. The specific activity of glutamic acid decarboxylase, the enzyme responsible for the synthesis of the neurotransmitter gamma-aminobutyric acid was high (10-30 nmol CO2/h/mg of protein) and electrophysiology showed that some cells had 'active' membranes. After about 18 months in culture, approximately 20% of the cells were able to respond to electrical stimulation by producing action potentials which were inhibited by 10(-7) M tetrodotoxin. These electrophysiological properties are stable: they have been repeatedly found at regular intervals throughout a 20 months period. Furthermore, a clone in which all tested cells are excitable, has been derived from the mass culture. Quail embryo neuroretina cells with typical neuronal properties can thus be established into permanent cultures after infection with Rous sarcoma virus.

Action Potentials↗
PubMed1983

A neuronal clone derived from a Rous sarcoma virus-transformed quail embryo neuroretina established culture.

Neuroretina (NR) is an evagination of the central nervous system (CNS) which is composed of photoreceptors, glial (Müller) cells and horizontal, bipolar, amacrine and ganglion neuronal cells. We describe here the usefulness of Rous sarcoma virus (RSV) in the establishment of a neuronal clone from quail embryo neuroretina. When primary cultures of chick and quail embryo neuroretina cells are transformed by RSV, neuronal markers such as ribbon synapses, choline acetyltransferase (CAT) and glutamic acid decarboxylase (GAD) specific activity are present. These RSV-transformed primary cultures can be established into permanent cell lines from which neuronal clones have been isolated. One of them, clone QNR/D, can generate tetrodotoxin(TTX)-inhibitable action potentials on electrical stimulation, has a high GAD activity and binds monoclonal antibodies raised against chick embryo neuroretina. The presence of these neuronal markers suggests that the QNR/D clone is derived from cells of the amacrine or ganglionic lineage. This is the first time that a neuronal cell clone of defined origin has been obtained from the CNS. The neuronal markers of the QNR/D clone are expressed at both the permissive and the non-permissive temperatures for transformation.

Action Potentials↗
PubMed1983

A glycine-enriched astrocytic cell clone derived from mouse cerebella transformed in vitro by simian virus-40.

Measurements were made of the amino acid content of a cellular clone (K55) derived from mouse cerebellar cultures transformed in vitro by simian virus-40 (Alliot and Pessac, 1981) and that appears to be astroglial. Both the total amount of amino acids as well as the percentage of glycine in K55 cells were higher than in the mixed cultures from which they are derived. Further, glycine accumulates in the culture medium of K55 cells, but not in the medium of the parental mixed cell culture (C14), thereby suggesting that glycine is synthesized and released by K55 cells.

Amino Acids↗
PubMed1982

[Immunoreactive neuropeptides in cell lines derived from nervous and non nervous tissues].

Immunoreactive neuropeptides have been found in cells of permanent lines originating from nervous (cerebellum, retina) and non nervous (mesenchyme, kidney, bladder, colon, lung, melanocyte) tissues. The pattern of neuropeptides varied from one cell type to another, but all the cell lines studied contained immunoreactive alpha-endorphin and methionin-enkephalin.

Animals↗
PubMed1982

Expression of neuronal markers in chick and quail embryo neuroretina cultures infected with Rous sarcoma virus.

Cultures of neuroretina (NR) cells from 7-day chick and quail embryos were infected with ts NY-68, a thermosensitive mutant of Rous sarcoma virus (RSV) which transformed NR cells at 36 degrees C. The following differentiation markers for neurones were studied: tetanus toxin-binding sites at the cell surfaces, presence of synapses, and the specific activity of the enzymes choline acetyltransferase (CAT) and glutamic acid decarboxylase (GAD). Appearance of synapses and expression of CAT were similar in control and transformed cultures. Tetanus toxin-binding cells were observed in transformed primary cultures and also in quail NR subcultures. GAD-specific activity was markedly stimulated in chick and quail primary cultures transformed by ts NY-68 and further increased in subcultures of ts NY-68-transformed quail NR cells. Stimulation of GAD activity is controlled by the transforming (src) gene of RSV since it was not observed in cultures infected with RAV-1, a leukosis virus which lacks the src gene. These data show that infection of chick and quail NR cultures with RSV results in the transformation of cells with neuronal markers.

Animals↗
PubMed1981

[Demonstration of neuropeptides in cultured neuroretinal cells of chick embryo].

Immunoreactive neuropeptides have been found in cultures of neuroretina cells obtained from 7 day Chick embryos. Methionine-enkephalin and thyroliberin (TRH) were present in a large number of neurons, while somatostatin, beta-melanotropin (MSH) and alpha-endorphin were occasionally seen in a few neurons. In cultures devoid of neurons and probably made of Müller (astroglial) cells, methionine-enkephalin, alpha-MSH and TRH were each detected in 60 to 80% of the cells. Immunoreactive methionine enkephalin was also localized in Müller cells in retina sections of 10 and 20 day old embryos.

Animals↗