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Biomedical subjects

F Gros

Publications and source records attributed to F Gros.

At least 109 records · Page 6Linked to original sources

Isolation of murine neuron-specific and non-neuronal enolase cDNA clones.

cDNA clones corresponding to subunits of neuron-specific (gamma gamma and alpha gamma) and non-neuronal (alpha alpha) enolase isozymes were characterized from two mouse brain cDNA libraries. Our hybridization data revealed a partial homology of the coding sequences of mouse alpha, mouse gamma and rat gamma mRNAs. The noncoding sequences, however, appear to be specific for each mouse mRNA. Although coding for two polypeptides of the same molecular weight, the mRNA for the gamma subunit (2600 bases) is larger than that for the alpha subunit (1900 bases). The noncoding sequences for neuron-specific gamma mRNA (about 1300 bases) are therefore longer than those of the non-nervous tissue specific alpha mRNA (about 600 bases).

Animals↗

Effects on mitochondrial metabolism of CCA, one inducer of neuroblastoma differentiation.

CCA, a potent neuroblastoma differentiation inducer, was shown by oxygraphic measurements to reduce significantly the O2 consumption of whole neuroblastoma cells as of mitochondria purified from neuroblastoma or mouse cortex. The effect of CCA on the respiration was compared to those of oligomycin. Our results suggest that the molecular target of CCA is the matrix F1 catalytic component of the F0F1 mitochondrial ATPase.

Animals↗

[Risk factors in gastric ulcer surgery after the age of 70 years. 34 cases].

Post-operative morbidity and mortality factors were investigated in a retrospective study of 34 patients aged over 70 operated upon for gastric ulcer. In two-thirds of the patients the operation was motivated by a complication of the ulcer. Two-thirds presented with an concurrent disease at the time of surgery. Fourteen patients died in hospital during the post-operative period, this mortality being related to the concurrent disease. Bleeding of the ulcer and emergency surgery were aggravating factors. The absence of post-operative abdominal complications had more influence on the prognosis than the type of operation performed.

Age Factors↗

Two simian virus 40 (SV40)-transformed cell lines from the mouse striatum and mesencephalon presenting astrocytic characters. I. Immunological and pharmacological properties.

Dissociate cultures were initiated from embryonic rostral mesencephalic and striatal tissues dissected from the mouse brain and previously incubated with a simian virus 40 (SV40) suspension. After several weeks in culture foci of fastly dividing cells were resuspended and cloned by successive dilutions. Several clones expressing the SV40 nuclear T antigen were obtained by these procedures and two of them, one mesencephalic (F7-Mes) and one striatal (F12-Str) were screened for the expression of glial or neuronal characters. Both clones possess adenylate cyclase-linked beta 2-adrenergic receptors. They also take up and synthesize gamma-aminobutyric acid (GABA) in amounts compatible with a glial origin. As is the case for astrocytes, the uptake of GABA is inhibited by beta-alanine and rather insensitive to the presence of diaminobutyric acid (DABA), a specific inhibitor of the neuronal GABA carrier. The most convincing evidence that F7-Mes and F12-Str belong to the astrocytic lineage comes from the fact that the two cell lines synthesize glial fibrillary acidic protein (GFAP) as demonstrated by immunofluorescence and immunoblotting. In an accompanying paper we also show that these lines behave like astrocytes when considered from the point of view of neuroglial interactions.

Animals↗

Two simian virus 40 (SV40)-transformed cell lines from the mouse striatum and mesencephalon presenting astrocytic characters. II. Interactions with mesencephalic neurons.

In an accompanying paper we report the characterization on the basis of pharmacological and immunological criteria of two astrocytic cell lines originating from the rostral mesencephalon and the striatum of the embryonic mouse (F7-Mes and F12-Str). This report compares the interactions of primary mesencephalic neurons with the astrocytic clones to that displayed with either an SV40-transformed fibroblastic clone (BT2) or primary mesencephalic (G-Mes) and striatal (G-Str) astrocytes. We show that BT2 differs from all other cell types (F7-Mes, F12-Str, G-Mes and G-Str). Indeed, as opposed to these cells BT2 is a poor substratum for neuronal adhesion or neuritic growth. This was clearly demonstrated by morphological examination of cocultures of the tested cells with either mesencephalic explants or dissociated cells. In addition a statistical analysis is provided which only concerns the dopaminergic (DA) neurons visualized by autoradiography after specific uptake of [3H]DA. The number of DA cells attached, the total length of their neurites and the degree of branching behaviour were examined. With the help of these criteria we show that F7-Mes and F12-Str are very similar to primary astrocytes and differ highly significantly from BT2. However, although sharing the main astrocytic features, F7-Mes and F12-Str do not differ from one another in their ability to induce the branching of DA neurites as their non-transformed counterparts do.

Animals↗

Differential expression of several neurospecific beta-tubulin mRNAs in the mouse brain during development.

The expression of alpha- and beta-tubulin mRNAs has been studied during mouse brain development by using in vitro translation and specific cDNA clone hybridization techniques. The results presented here indicate that about half of the isotubulins expressed in vivo by brain cells are directly encoded by mRNA, the other isotypes being of post-translational origin. Among the six brain beta-tubulin mRNAs detected after RNA size fractionation followed by in vitro translation, four (1.8a, 1.8c, 2.4 and 3.5 kb) appear to be uniquely expressed in brain, and very likely in neurons. These four beta-tubulin RNA transcripts are differently regulated during development. The 1.8a-kb mRNA, which codes for the neurospecific beta' 1-isotubulin, is expressed very early, whereas the 1.8c, 2.4 and 3.5-kb mRNAs, which all code for a beta 4-tubulin isotype, are all expressed later. We isolated a beta-tubulin cDNA clone which specifically hybridizes to the large, neural 3.5-kb mRNA. The two other ubiquitous tubulin mRNAs, 1.8b and 2.9 kb in length, both direct the synthesis of an electrophoretically distinct beta 3 isotype. These results support the idea that within the tubulin multigene family, distinct beta-tubulin isogenes could be specifically expressed at various steps of the neuronal differentiation, thus contributing to the isotubulin diversity and specificity observed in this cell type and possibly involved in the developmental changes of its structure and function.

Animals↗

[Copurification of tyrosine hydroxylase from rat pheochromocytoma by protein kinase].

Rat pheochromocytoma contains a protein kinase activity which remains associated with tyrosine hydroxylase (TH) during its purification. The incorporation of phosphate in TH is observed after incubation of TH with labelled ATP and magnesium without the need for an exogenous protein kinase. This Ca2+ and cAMP-independent kinase activity is different from previously described TH phosphorylating kinases from rat pheochromocytoma and other tissues.

Adrenal Gland Neoplasms↗

[Nuclear proteins preferentially bound to the beta-globin gene: cellular specificity and variation during terminal differentiation of mouse erythroblasts].

Restriction fragments of the mouse beta major globin gene and of the long terminal repeat (LTR) DNA fragment of the mouse mammary tumor provirus as a control, were used to analyze the specificity of DNA-protein interactions in nuclear extracts of mouse erythroleukemia (MEL) cells and of other differentiated mouse cultured cell lines. After gel electrophoresis and transfer to nitrocellulose, DNA-binding proteins with a preferential affinity for the cloned beta-globin genomic sequence were characterized and related to the level of globin gene expression during induction of differentiating mouse erythroblasts. Two proteins (110 K and 75 K) appear in differentiated MEL cells while another one (100 K), for which we have localized the binding site on the beta-globin gene, is present only in immature MEL cells.

Animals↗

Correlation of bronchial epidermoid metaplasia with level of tobacco consumption in heavy smokers.

One hundred forty-four heavy-smoker (125 males, 19 females) volunteers with at least 15 packet-year of smoking experience (number of daily packets of cigarettes X number of years of smoking) underwent bronchoscopy with systematic biopsies in ten sites of the bronchial tree. No morbidity was related to the fibroscopy procedure. Each biopsy was cut into ten sections and an index of metaplasia (IM) was calculated: IM = number of sections with epidermoid metaplasia/number of sections examined X 100. A highly significant statistical correlation was observed (P = 10(-4)) between the IM and the amount of cigarettes smoked (slope of the curve: 0.23). Sex appeared to play an important role in the incidence of metaplasia, since 84% of the examined females had an IM less than 15% versus 48% of the males (P adjusted for individual tobacco consumption in packet-years less than 0.01). Early study of HLA phenotype (locus A) failed to detect a link between HLA and risk of tobacco-induced epidermoid metaplasia.

Epidermal Cells↗

In vitro translation of human pheochromocytoma messenger RNAs: characterization of tyrosine-hydroxylase and dopamine-beta-hydroxylase.

mRNAs extracted from human pheochromocytoma were translated in vitro in a lysate of a rabbit reticulocytes. Two enzymes of the biosynthetic pathway of the catecholamines, tyrosine-hydroxylase (TH) and dopamine-beta-hydroxylase (DBH), were characterized as translation products after immunoprecipitation by specific antisera and electrophoretic analysis. The precursor of TH is a polypeptide having a molecular mass of 62,000 identical to that found for the mature protein. The molecular mass of the precursor of DBH 73,000 while that of the mature form is 79,000. TH and DBH have been translated from mRNAs having sedimentation coefficients of 22S and 25S, respectively.

Adrenal Gland Neoplasms↗

Changes in mitochondrial proteins during neuroblastoma differentiation.

The evolution of three major mit-proteins was followed in neuroblastoma cells cultured in different conditions of differentiation. 1 methyl cyclohexane carboxylic acid (CCA) was found to stimulate the synthesis of the three mit-protein markers. This result, compared to the effects of oligomycin, an inhibitor of mitochondrial function, favours the hypothesis that CCA induces in vitro neurogenesis through a general metabolic alteration.

Animals↗

[Microheterogeneity of tubulin in the central nervous system of the quail (Coturnix coturnix japonica)].

Tubulin from the central nervous system of the Quail (Coturnix coturnix japonica) was analysed by two-dimensional gel electrophoresis. Either in soluble extracts or in tubulin enriched fractions, tubulin of forebrain, cerebellum and spinal cord is resolved into 3 components alpha, beta' and beta, characterized by their electrophoretic coordinates (alpha: pI = 5.4, MW = 54 kd, beta: pI = 5.2, MW = 52 kd; beta': pI = de mapping, by comparison with mouse brain alpha, beta and beta'-tubulin components. With highly resolutional isoelectric focusing 6 alpha isotubulins and 14 beta isotubulins are found in enriched-tubulin fractions of Quail forebrain and cerebellum.

Animals↗

Tissue-specific formation of transcription-initiation complexes at the 5' end of the mouse beta major globin gene.

Accurate initiation of transcription in vitro requires, in addition to RNA polymerase II, factors present in soluble extracts of cultured cells. We have developed transcription system in vitro, which permits us to visualize the transcription-initiation complexes on a mouse beta globin restricted fragment from a recombinant beta globin bacteriophage DNA. Using the lambda fragments as internal controls this system has allowed us to assess the specificity of transcription with RNA polymerase II. Comparing extracts from cells and tissues expressing the globin genes (Friend cells, spleen and blood from phenylhydrazine-induced anemic mice) with those which do not (thymus, liver, PCC3 cells), we observed that specific initiation of transcription on the beta globin gene occurs only with soluble extracts from erythroid tissues. This tissue-specific transcription is partially sensitive to alpha-amanitin and occurs at the 5' end of the globin gene.

Amanitins↗

Developmental changes in translatable mRNAs for the cerebral enolase isozymes alphaalpha and gammagamma.

Using the rabbit reticulocyte cell-free translation system we have estimated during ontogenesis the proportions of in vitro translatable alpha and gamma brain enolase mRNAs, which are two minor mRNA species. No polypeptide precursor to these enzyme subunits appears to be synthesized during translation in vitro. During brain development, the changes in translatable alpha and gamma mRNA content seem to parallel those of the corresponding antigens. The proportion of each of the enolase mRNAs is highest in adult mouse brain. Mechanisms controlling alpha and gamma antigen expression are discussed. In order to prepare the specific cDNA probes, purification of alpha and gamma mRNAs was undertaken.

Animals↗

Adverse effect of indomethacin in experimental chronic nephrosis.

The effects of long-term indomethacin therapy were studied in rats with chronic nephrosis induced by puromycin of aminonucleoside (PAN). Fifteen PAN rats received indomethacin (2 mg/kg/d) administered by daily gavages, and fifteen received glucose. A control group received indomethacin but no PAN. Three to five animals per group were sacrificed monthly. Proteinuria was similar in both PAN rat groups. Renal function and histology were normal in the control rats. In the PAN animals, renal failure developed earlier in the treated rats than in the others. Histological examination revealed in all PAN rats early tubulo-interstitial lesions and progressive glomerular damage. The pattern of the lesions was identical in both PAN rats, but the glomerular changes developed earlier, were more diffuse and more severe in the indomethacin treated rats. Thus, indomethacin resulted in an accelerated destruction of the glomeruli in experimental nephrosis. Such a deleterious effect remains unexplained, and its relevance to human disease is not demonstrated.

Animals↗

[Proliferation and differentiation of a myogenic line in synthetic media].

When plated in plastic culture dishes, muscle cells of the L6 cell line are unable to proliferate or to differentiate in a synthetic medium composed of Dulbecco's Minimal Essential medium (DME), bovine fetuin (1 mg/ml) and bovine insulin (10 micrograms/ml). In contrast, these cells will proliferate and differentiate morphologically and biochemically in this medium if the culture dishes have been previously treated with DME containing fetal Calf serum. Pretreatment of dishes does not increase cell attachment in synthetic medium but seems to act as a factor increasing cell viability.

Animals↗