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E Perret

Publications and source records attributed to E Perret.

At least 37 records · Page 2Linked to original sources
PubMed2000

Identification of rpaP1-5 and rpaP2-6 genes encoding two additional variants of the 60S acidic ribosomal proteins of Schizosaccharomyces pombe.

In the fission yeast, four genes (rpaP1-1, rpaP1-3, rpaP2-2, and rpaP2-4) encoding two variants of the RpaP1 and RpaP2 ribosomal proteins (rp) have been characterized. We have identified cDNA for additional variants called RpaP1.5 and RpaP2.6. Sequence comparison suggests that RpaP1.5 diverged before RpaP1.1 and RpaP1.3 and that RpaP2.6 is closer to RpaP2.2 than to RpaP2.4. The corresponding genes, rpaP1-5 and rpaP2-6, are transcribed coordinately with other rp genes.

Amino Acid Sequence↗
PubMed2000

Differential expression and cellular distribution of centrin isoforms during human ciliated cell differentiation in vitro.

Centrin protein is an ubiquitously expressed cytoskeletal component and is a member of the EF-hand superfamily of calcium-binding proteins. It was first discovered in the flagellar apparatus of unicellular green algae where it is involved in contraction of Ca(2+)-sensitive structures. Centrin protein is associated with centrosome-related structures such as spindle pole body in yeast, and centriole/basal bodies in flagellar and ciliated cells. Three centrin genes have been cloned in human cells. In this work, we have performed a comparative biochemical and functional analysis of centrin isoforms using a primary culture of human nasal epithelial cells which provides an efficient way to obtain a complete ciliated cell differentiation process. RT-PCR experiments show that the expression of the three human centrin genes increases during cell differentiation, and that only centrin 2 and 3 are expressed during cell proliferation. Using polyclonal antibodies raised against recombinant human centrin 2 and 3, we show a specific pattern of protein expression. Ultrastructural immunolocalization suggests that centrin proteins are involved in the early process of centriole assembly, as they are concentrated within the precursor structures of centriole/basal bodies. It also shows a differential localisation of centrin proteins in mature centriole/basal bodies, suggesting different functions for centrins 1/2 and centrin 3. This is also supported by functional analyses showing that centrin 1 and/or centrin 2 are involved in ciliary beating.

Calcium-Binding Proteins↗
PubMed1999

Entry of human cytomegalovirus into retinal pigment epithelial and endothelial cells by endocytosis.

PURPOSE: Human retinal pigment epithelial (RPE) cells and endothelial cells (HUVECs) are targets of human cytomegalovirus (HCMV) infection in vivo with significantly protracted replication in vitro compared with that in fibroblasts. This study analyzes the kinetics and mechanisms of HCMV entry into both cell types. METHODS: RPE cells were obtained from donor eyes. HUVECs were isolated from human umbilical cords. HCMV entrance was followed by electron microscopy and immunofluorescence in the presence of lysosomotropic agents and cytochalasin B. RESULTS: Human cytomegalovirus entered into RPE cells and HUVECs as early as 5 minutes after virus- cell contact. Entry was mediated by endocytosis, whereas HCMV enters fibroblasts through fusion. Most internalized viral particles and dense bodies appeared to be degraded within vacuoles. Viral entry, transport of viral proteins to the nucleus, and onset of viral transcription (immediate early [IE] protein expression) were significantly blocked by cytochalasin B. Lysosomotropic agents did not significantly reduce IE expression in RPE cells or HUVECs. CONCLUSIONS: This study shows that HCMV penetrates these highly specialized relevant cells via endocytosis. The low level of infection and the delay in the onset of HCMV expression seen in these cells compared with fibroblasts may be related to the sequestration and degradation of incoming viral particles in endocytic vacuoles.

Ammonium Chloride↗
PubMed1998

Improved differential screening approach to analyse transcriptional variations in organized cDNA libraries.

A cDNA library was generated from rat brain tissues and organized into 1536-well plates, using a fluorescence activated cell sorter (FACS), acting as a single cell deposition system. The organized library containing 10,000 clones, with 60% full-length cDNA inserts, allowed the generation of multiple identical membrane replicas. Each replica was hybridized with a complex probe obtained from a particular brain tissue or a given cultured cell. The signal intensity for each of the clones present on the membrane, quantified with a standard image-analysis software, is proportional both to the abundance of the corresponding mRNA in the probe and to the amount of plasmid template on the membrane. The latter value was thus used to normalize the signals produced with complex probes, to optimize the comparison of mRNA expression levels for the different systems under study. The construction of high-quality cDNA libraries, the generation of identical membrane replicas and comparable probes, and the utilization of an image-analysis software package, coupled with the normalization of the spot intensity by assaying plasmid quantity, significantly improves the differential screening approach. Altogether, these technical improvements open the possibility to compare a great number of different probes and, in consequence, to accumulate biological information for each clone present in an organized cDNA library. The functional information obtained should complement data from DNA sequencing projects.

Animals↗
PubMed1998

The cell cycle gene SKP1 is regulated by light in postnatal rat brain.

During the early postnatal phase of high neuronal plasticity, an altered visual input leads to great modifications of visual cortex organization [Y. Frégnac, M. Imbert, Development of neuronal selectivity in primary visual cortex of cat, Physiol. Rev., 64 (1984) 375-434; D.H. Hubel, T.N. Wiesel, S. LeVay, Plasticity of ocular dominance columns in monkey striate cortex, Philos. Trans. R. Soc. London, Ser. B, 278 (1977) 377-409.]. We used refined differential screening of an organized cDNA library to identify the genes that may participate in this plasticity. We isolated a candidate plasticity gene encoding for a 163 aa protein that is closely related to the human and yeast Skp1p, a key factor in cell cycle progression [C. Baï, K. Hofman, L. Ma, M. Goebl, J.W. Harper, S.J. Elledge, SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box, Cell, 86 (1996) 263-274; C. Connelly, P. Hieter, Budding yeast SKP1 encodes an evolutionary conserved kinetochore protein required for cell cycle progression, Cell, 86 (1996) 275-285; H. Zhang, R. Kobayashi, K. Galaktionov, D. Beach, p19Skp1 and p45Skp2 are essential elements of the cyclin A-CDK2 S phase kinase, Cell, 82 (1995) 915-925.]. Northern blot analysis showed that the expression of SKP1 (Skp1p gene) dramatically decreased after 2 h of light stimulation in the visual cortex of young dark-reared rats. This down regulation lasted at least 72 h. It was specific for the critical period as we did not observe any significant regulation of SKP1 mRNA by light in adult dark-reared rat brain. The down regulation was observed in the superior colliculus but also in the frontal cortex and in the hippocampus. The fact that this down regulation was not restricted to the visual system, suggested that it could be produced by dark rearing-induced hormonal changes. The significance of SKP1 expression in the brain and its regulation are discussed.

Animals↗
PubMed1998

Differentially expressed genes in C6.9 glioma cells during vitamin D-induced cell death program.

C6.9 rat glioma cells undergo a cell death program when exposed to 1, 25-dihydroxyvitamin D3 (1,25-D3). As a global analytical approach, we have investigated gene expression in C6.9 engaged in this cell death program using differential screening of a rat brain cDNA library with probes derived from control and 1,25-D3-treated cells. Using this methodology we report the isolation of 61 differentially expressed cDNAs. Forty-seven cDNAs correspond to genes already characterized in rat cells or tissues. Seven cDNAs are homologous to yeast, mouse or human genes and seven are not related to known genes. Some of the characterized genes have been reported to be differentially expressed following induction of programmed cell death. These include PMP22/gas3, MGP and beta-tubulin. For the first time, we also show a cell death program induced up-regulation of the c-myc associated primary response gene CRP, and of the proteasome RN3 subunit and TCTP/mortalin genes. Another interesting feature of this 1,25-D3 induced-cell death program is the down-regulated expression of transcripts for the microtubule motor dynein heavy chain/MAP 1C and of the calcium-binding S100beta protein. Finally 15 upregulated cDNAs encode ribosomal proteins suggesting a possible involvement of the translational apparatus in this cell program. Alternatively, these ribosomal protein genes could be up-regulated in response to altered rates of cellular metabolism, as has been demonstrated for most of the other isolated genes which encode proteins involved in metabolic pathways. Thus, this study presents to our knowledge the first characterization of genes which are differentially expressed during a cell death program induced by 1, 25-D3. Therefore, this data provides new information on the fundamental mechanisms which participate in the antineoplastic effects of 1,25-D3 and on the machinery of a cell death program in a glioma cell line.

Animals↗
PubMed1997

Expression of SKP1 mRNA and protein in rat brain during postnatal development.

We previously isolated the cell cycle element SKP1 as a candidate plasticity gene in the rat visual cortex. Here, we studied the expression and localization of SKP1 mRNA and protein in rat brain. We found a high level of expression for the SKP1 gene in the cortex at different postnatal ages. SKP1 mRNA levels remained stable from P2 (postnatal day 2) to adulthood. SKP1 mRNA expression was also detected in several other brain areas. Skp1p (SKP1 protein) immunohistochemistry showed nuclear staining in a large majority of neurones. The pyramidal cells in the hippocampus, as well as cortical cells were stained. The presence of Skp1p in post-mitotic neurones suggests that this protein is involved in processes other than the cell cycles other target proteins and functions in neurones are currently under investigation.

Animals↗
PubMed1995

Identification of an HSP70-related protein associated with the centrosome from dinoflagellates to human cells.

The monoclonal antibody CTR210 raised against isolated human centrosomes strongly decorates the centrosome and more weakly a domain congruent with the Golgi apparatus in several animal cells (HeLa, 3T3, CHO, PtK2). Both decorations resist Triton extraction in conditions which totally extract the Golgi apparatus, as judged by galactosyltransferase decoration. A 67 kDa centrosomal antigen can be demonstrated in human cells with this antibody. CTR210 also decorates the centrosome or associated structures in several systems, including unicellular eukaryotes such as dinoflagellates or ciliates. A 72 kDa antigen has been identified and purified from the dinoflagellate C. cohnii and its NH2-terminal sequence partially established. It shows a close homology with HSP70 proteins. The possibility that the 72 kDa antigen belongs to this chaperone family was further supported using a mAb reacting, in most species, with HSP70. A polyclonal antibody raised against the 72 kDa antigen from C. cohnii decorates the centrosome in human cells and reacts with the CTR210 centrosomal 67 kDa antigen. These results suggest that specific chaperone proteins are associated with the centrosome in eukaryotic cells. The centrosomal chaperones could participate in the microtubule nucleation reaction or in the process of centrosome assembly.

Amino Acid Sequence↗
PubMed1993

Fluorescence induction used to measure parathion toxicity in the marine dinoflagellate Prorocentrum micans: a possible model for biodetection.

The marine dinoflagellate Prorocentrum micans Ehrenberg was used as a test organism to determine the conditions of use of fluorescence induction kinetic measurements in the study of parathion phytotoxicity. Measurements were taken of the kinetics of slow and fast fluorescence induction in whole cells and isolated chloroplast fragments at various concentrations of parathion. In both types of induction, the addition of parathion greatly decreased the fluorescence yield, indicating either inhibition of electron transport or physical changes. The action of parathion was a function of its concentration and was similar to the action of DCMU. Fluorescence induction was clearly less in isolated chloroplast fragments than in whole cells.

Animals↗
PubMed1993

Microtubule organization during the cell cycle of the primitive eukaryote dinoflagellate Crypthecodinium cohnii.

The complete microtubular system of the dinoflagellate Crypthecodinium cohnii Biecheler is described, as seen by confocal laser scanning fluorescence microscopy and labelling with anti-beta-tubulin antibody. This technique allowed us to observe the organization of the subcortical and internal cytoskeletons and the mitotic microtubular system, and their changes during the cell cycle. These observations are compared with those made in cryosections by light microscopy and in fast-freeze-fixed, cryosubstituted cells by electron microscopy. We show the organization of the cortical microtubules, and in particular of the thick microtubular bundles arranged as a three-pronged fork from which they seem to emanate. This fork emerges from a peculiar cytoplasmic zone at the pole of the cell and is in contact with the region of the kinetosomes, at the cingulum. During the G1 phase, only a single, radial microtubular bundle (a "desmose") is observable in the inner part of the cytoplasm. One of its ends is near the flagellar bases and the other end is close to the nucleus in the centrosome region. During the S phase, the flagella drop off, the cell encysts and the kinetosomes duplicate. In mitosis, the cortical microtubules and the intracytoplasmic microtubular bundles do not depolymerize. The microtubular fork, desmose and centrosome double and migrate, while the divided kinetosomes stay in the same place. Later, the centrosomes organize the extranuclear spindle, which is connected to the kinetosome region by the microtubular desmose. The convergent end of the three-pronged fork seems to be in contact with the centrosome region. In early and mid-prophase, thick microtubular bundles pass through the nucleus in cytoplasmic channels and converge towards the two poles. Asters were never seen at the spindle poles. The channels and microtubular bundles in the spindle double in number during late prophase and lengthen in early anaphase. The spindle bundles diverge in late anaphase, extend to very near the plasma membrane and depolymerize during telophase. The cleavage furrow in which tubulin and actin are characterized appears in anaphase, formed by invagination of plasma membrane in the kinetosome region. The structure and rearrangements of the Crypthecodinium cohnii microtubular system are compared with those of other dinoflagellates and protists and of higher eukaryotes.

Animals↗
PubMed1991

Microtubular spindle and centrosome structures during the cell cycle in a dinoflagellate Crypthecodinium cohnii B.: an immunocytochemical study.

In order to determine if a mitotic spindle organizing center is present in dinoflagellate cells, we used a library of 12 monoclonal antibodies obtained by immunizing mice with isolated human centrosomes. When tested by immunofluorescence on cryosections of the dinoflaggelate Crypthecodinium cohnii B., a positive labeling was obtained with three of these antibodies. In interphase cells, the anti-centrosome antibodies labeled structures located either in the cell periphery, corresponding probably to both basal bodies (i.e. kinetosomes) and in the perinuclear area. In the latter case, two punctate structures were observed near the nuclear envelope. They have never been described, either in light, or in electron microscopic studies of dinoflagellates. We have designated them as centrosome-like structures. A microtubular desmose reacting positively with anti-tubulin Ab was also visible, linking kinetosomes and centrosome-like structures. During mitosis, the double punctate structures were observed at the poles of the nucleus. Double immunolabeling with tubulin and anti-centrosome Ab was also carried out and strongly suggested that in mitotic cells, centrosome-like structures, located at the poles of the mitotic spindle, were associated with microtubular bundles and probably organize and polarize them. These data indicate the existence of centrosome-like structures in C. cohnii cells and the strong conservation of some centrosomal epitopes from dinoflagellates to human. One of the antibodies (CTR 210) recognized by immunoblotting, a single protein band at 72 kDa from a total protein extract. The direct demonstration that this protein is located at the centrosome-like structures and at the kinetosomes deserves further study.

Animals↗
PubMed1988

Role of interstimulus and stimulus-hemisphere compatibility in the process of interhemispheric integration.

Normal subjects had to name German compound nouns which were presented tachistoscopically. The compound nouns were displayed either unilaterally to the left or right visual field, or bilaterally with one element to the left and one to the right visual field. A distinction was made between the bilateral conditions as to whether the representation of the elements, printed and/or pictorial, included a high or low interstimulus and a high or low stimulus-hemisphere compatibility. Analysis indicated firstly a superiority of the left hemisphere for the naming of compound nouns in mixed print and pictorial representation. Secondly, the performance in the bilateral conditions was moderated by stimulus-hemisphere compatibility. In the process of interhemispheric integration stimulus-hemisphere compatibility proved more crucial than interstimulus compatibility. Analyses of errors further illustrated hemispheric behaviour.

Adult↗
PubMed1987

Interhemispheric integration of compound nouns: effects of stimulus arrangement and mode of presentation.

Normal subjects had to name German compound nouns which were presented tachistoscopically. The compound nouns were displayed either unilaterally to the left or right visual field or bilaterally with one element to each visual field. In the bilateral condition a distinction was made as to whether familiar or unfamiliar arrangement of the elements was used. Representation in print was compared with pictorial representation of the compound nouns. A right visual-field superiority was observed with printed representation, but no laterality effects with pictorial representation. Bilateral processing was superior to unilateral processing. Within the bilateral conditions, the familiar arrangement of the elements yielded a significantly better performance than unfamiliar arrangement. This difference can be explained by reading habits and/or by different styles of interhemispheric integration.

Adult↗
PubMed1986

Interhemispheric integration of information in a surface estimation task.

Normal subjects were asked to estimate the total surface of two black spots presented tachistoscopically, either both to the right or both to the left visual field, or bilaterally to either visual field. Performance was highest with unilateral left visual field presentation and lowest with bilateral presentation of the spots. Interhemispheric integration varied with the quantitative relation between the stimuli in the left and right visual fields. Errors may be due in part to an insufficient cooperation of the left hemisphere and/or to an ignorance of the right hemisphere and/or to an unreliable transfer by the cerebral commissures.

Adult↗
PubMed1986

A pronounced cognitive style improves interhemispheric integration: by hemispheric differentiation or by hemispheric similarity?

Under conditions of tachistoscopic stimulation normal subjects were presented visuospatial stimulus pairs which had to be integrated interhemispherically. The presence of a pronounced cognitive style improves the interhemispheric integration of this type of information. Two interpretations are discussed. The first explains the observed improvement with a qualitative division of labour, with one highly specialized and one just transmitting hemisphere, i.e., with hemispheric differentiation. The second emphasizes the relatively high capacity of the "subordinate" hemisphere in information processing, i.e., hemispheric similarity.

Adult↗