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E Couve

Publications and source records attributed to E Couve.

12 recordsLinked to original sources

Intracellular distribution of myelin protein gene products is altered in oligodendrocytes of the taiep rat.

Hypomyelination and subsequent demyelination of the taiep rat CNS are thought to result from the abnormal accumulation of microtubules (MTs) in oligodendrocytes that disrupts intracellular transport of components needed to form and maintain the myelin sheath. In this study, myelin gene expression was evaluated in mutant and age-matched controls to determine if MT abnormalities affect the distribution of myelin proteins and their mRNAs. Immunohistochemical analysis of taiep brains and spinal cords revealed a gradual decrease in levels of several myelin proteins including myelin basic protein (MBP), proteolipid protein (PLP), myelin-associated glycoprotein (MAG), and 2',3'-cyclic nucleotide 3'-phosphodiesterase. Accompanying early declines in MAG and PLP, accumulations of immunoreactive products were detected within oligodendrocytes, consistent with a defect in protein trafficking. Northern blot analysis indicated that diminishing protein levels could not be attributed to changes in transcriptional activity, except for MBP of which mRNA levels decreased with age. Cellular localization of MBP mRNA by in situ hybridization further revealed that transcripts were concentrated within oligodendrocyte cell bodies instead of uniformly distributed throughout processes. These results demonstrate that changes in expression and intracellular localization of myelin gene products are concurrent with increases in MT mass in taiep oligodendrocytes and support our hypothesis that cytoskeletal defects prevent the normal transport of elements required for the formation and maintenance of the myelin sheath.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

The 102-kilobase pgm locus of Yersinia pestis: sequence analysis and comparison of selected regions among different Yersinia pestis and Yersinia pseudotuberculosis strains.

We report the complete 119,443-bp sequence of the pgm locus from Yersinia pestis and its flanking regions. Sequence analysis confirms that the 102-kb unstable pgm locus is composed of two distinct parts: the pigmentation segment and a high-pathogenicity island (HPI) which carries virulence genes involved in iron acquisition (yersiniabactin biosynthetic gene cluster). Within the HPI, three genes coding for proteins related to phage proteins were uncovered. They are located at both extremities indicating that the entire HPI was acquired en bloc by phage-mediated horizontal transfer. We identified, within the pigmentation segment, two novel loci that may be involved in virulence: a fimbriae gene cluster and a locus probably encoding a two component regulatory system similar to the BvgAS regulatory system of Bordetella pertussis. Three genes containing frameshift mutations and two genes interrupted by insertion element insertion were found within this region. To investigate diversity among different Y. pestis and Yersinia pseudotuberculosis strains, the sequence of selected regions of the pgm locus and flanking regions were compared from 20 different Y. pestis and 10 Y. pseudotuberculosis strains. The results showed that the genes interrupted in Y. pestis are intact in Y. pseudotuberculosis. However, one of these mutations, in the bvgS homologue, is only present in Y. pestis strains of biovar Orientalis and not in those of the biovars Antiqua and Medievalis. The results obtained by analysis of variable positions in the sequence are in accordance with historical records, confirming that biovar Orientalis is the most recent lineage. Furthermore, sequence comparisons among 29 Yersinia strains suggest that Y. pestis is a recently emerged pathogen that is probably entering the initial phase of reductive evolution.

Bacteriophages↗

Dysmyelination, demyelination and reactive astrogliosis in the optic nerve of the taiep rat.

Taiep is an autosomal recessive mutant rat that shows a highly hypomyelinated central nervous system (CNS). Oligodendrocytes accumulate microtubules (MTs) in association with endoplasmic reticulum (ER) membranes forming MT-ER complexes. The microtubular defect in oligodendrocytes, the abnormal formation of CNS myelin and the astrocytic reaction were characterized by immunocytochemical and ultrastructural methods during the first year of life. Optic nerves of both control and taiep rats were processed by the immunoperoxidase method using antibodies against tubulin, myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP). Taiep oligodendrocytes are strongly immunoreactive against tubulin, indicative of a significant accumulation of microtubules. Early differentiated oligodendrocytes observed with electron microscopy show that MT-ER complexes are mainly present in the cell body. This defect increases during the first year of life; oligodendrocytes show large MT-ER complexes projected within oligodendrocyte processes. Using anti-MBP, there was a progressive reduction of immunolabeling in the myelin sheaths as taiep rats grew older. Ultrastructural analysis revealed severely dysmyelinated axons with a frequently collapsed periaxonal collar. However, through age the myelin sheath became gradually infiltrated by MTs, suggesting their contribution to premature loss of myelin in the taiep rat. Axons of one-year-old taiep rats were severely demyelinated. Modifications in astrocytes revealed by the GFAP antibody showed a strong hypertrophy with increased immunostaining in their processes. As demyelination of axons progressed, taiep rats developed a strong astrogliosis. The present findings suggest that in taiep rats the early abnormal myelination of axons affects the adequate maintenance of myelin, leading to a progressive loss of myelin components and severe astrogliosis, features that should be considered in the pathogenesis of dysmyelinating diseases.

Animals↗

Binding of microtubules to transitional elements in oligodendrocytes of the myelin mutant taiep rat.

The presence of microtubules physically bound to smooth endoplasmic reticulum profiles of oligodendrocytes constitutes the most conspicuous feature observed in the myelin mutant taiep rat. The endoplasmic reticulum membranes associated with microtubules were morphologically characterized as transitional elements that constitute the intermediate compartment according to their topographic location close to the cis-Golgi apparatus. The development of this surprising microtubular defect appears to be associated with the early events of myelination. Transitional elements associated with microtubules operate in protein transport from endoplasmic reticulum to cis-Golgi. This microtubular defect could explain the dysmyelination and neurologic alterations observed in taiep rats. Moreover, these findings allow us to propose that there is a blockage of protein traffic at the intermediate compartment of taiep oligodendrocytes, a situation that could explain the hypomyelinated axons observed in this myelin mutant. The binding of microtubules to membranous organelles promotes the stabilization of microtubules, a feature that has important implications regarding its accumulation within the cytoplasm of oligodendrocytes during the temporal evolution of this neurologic disorder. The taiep rat is a myelin mutant with a long survival and could be a useful model for understanding dysmyelinating diseases in which the intracellular transport of myelin components is affected.

Animals↗

Infectious pancreatic necrosis virus internalization and endocytic organelles in CHSE-214 cells.

The early events that take place during the internalization of infectious pancreatic necrosis virus (IPNV) into Chinook salmon embryo cells (CHSE-214) were analyzed ultrastructurally. Endocytic tracers were employed in order to characterize the organization of endocytic organelles in CHSE-214 cells, as well its relation to the IPNV penetration. Results demonstrate that IPNV appear internalized within vesicular compartments which are located peripherally in CHSE-214 cells. Despite the high rate of infectious multiplicity few virus particles were detected inside the cells. Endocytic tracer labelling of tubulovesicular elements and endosomes of host cells showed a well developed endocytic apparatus. Results suggest that endocytosis may be involved during the initiating events in the productive IPNV infection.

Animals↗

Cell polarity changes and migration during early development of the avian peripheral auditory system.

The intracellular location of organelles was studied in avian embryos immediately before and during the initial detachment of cells from the dorsoanterolateral wall of the otocyst, using light and electron microscopy. The Golgi apparatus was silver-impregnated, and its location within the otic epithelium was determined quantitatively. The present study demonstrates that a sub-population of cells of the dorsoanterolateral otic epithelium changes the intracellular location of its organelles, particularly the Golgi apparatus and the centrosome, from an apical to a basal position. Concomitantly, cells lose specializations characteristically present at apical (tight junctions, microvilli) and basal (basal lamina) surfaces. At basolateral cell surfaces, filopodia form ahead of the Golgi apparatus and centrosome and penetrate the previously continuous underlying basal lamina. Thereafter, cells detach from the otocyst and migrate medially toward the hind-brain. Thus, concomitant with changes in surface polarity, the cells that comprise the dorsoanterolateral wall of the otocyst undergo profound changes in the intracellular location of their organelles, especially the Golgi apparatus and the centrosome, so that by the time cells detach from the otic epithelium a reversal in their "normal" internal polarity has occurred. We suggest that the change in cell polarity may be related to the mechanisms that allow cells to leave the otocyst.

Animals↗

Changes in the predentin thickness and mineralization front configuration in developing human premolars.

The thickness of the predentin layer was studied at three different levels of developing human premolars. The results demonstrate that at the growing end next to the apex, where dentinogenesis is most active, predentin exhibits its greatest thickness (mean value 40.4 micron). However, at the coronal region, where primary dentin has been completely formed, predentin width is reduced to a mean value of 14.8 micron. Changes in the calcospheritic configuration of the mineralization front were established for each of the predentin levels studied. A comparative analysis of these calcospheritic changes and the morphology of fluorescent tetracycline lines detected in ground sections of premolars was established. Fluorescent lines observed at the coronal circumpulpal dentin showed large calcospheritic forms beneath the mantle dentin. However, lines found near the dentin-pulp border showed small calcospherites. It is concluded that the thickness of the predentin layer and the mineralization front configuration vary as a function of dentinogenic activity during development of human premolars.

Bicuspid↗

Ultrastructural changes during the life cycle of human odontoblasts.

In developing premolars there are four successive stages of odontoblasts related to their location within the tooth, namely pre-odontoblasts, secretory, transitional and aged odontoblasts. Pre-odontoblasts are at the growing tip of the root; they are immature and slightly polarized. They become fully differentiated to give rise to the most active, secretory stage when primary dentine is produced. After completion of primary dentine formation in the coronal region, the aged odontoblasts have a reduced number of organelles which are relocated within the infranuclear region. This final stage is regressive and results from an autophagic process that is prominent in the transitional stage.

Bicuspid↗

Morphometric analysis of the nucleolus during the life cycle of human odontoblasts.

Developing first premolars were used as a model system to obtain information on the nucleolar structure of human odontoblasts at several stages of their life cycle. Four stages were defined by their location within the tooth: a) preodontoblasts were located at the growing tip of the root; b) secretory odontoblasts in the apical region; c) transitional odontoblasts in the middle region; and d) aged odontoblasts in the coronal region. Preodontoblasts have a small nucleolus (0.55 micron 2) with few strands of dense fibrillar material radiating from the fibrillar center. Secretory odontoblasts are characterized by a large (1.24 micron 2), irregular, and reticulated nucleolus. The fibrillogranular material, the largest component in all nucleoli, reaches maximal size at this stage (0.88 micron 2). Fibrillar centers occupy about the same area (0.1 micron 2) throughout the odontoblast's life cycle. As the formation of primary dentin is completed, the nucleolus of transitional odontoblasts is reduced in size (0.54 micron 2). Finally, the aged odontoblasts have a small, compact nucleolus (0.39 micron 2), with segregated components. Morphologic analysis and quantification of size and component areas of nucleoli obtained with an image analyzer indicated that secretory odontoblasts had the most active, and aged odontoblasts the least active, nucleolus.

Bicuspid↗

Analysis of pore-rich areas on the nuclear envelope of spermatocytes.

Serial sections analysis of spermatocytes of the rodent Phyllotis darwini, reveal a persistent positional relationship between nucleolar material and centrioles. Pore rich areas interposed between these nucleocytoplasmic components, are represented graphically. Centriole replication takes place during the pachytene stage. Pore rich areas could be the result of a preferential aggregational process which insures the exchange of information between nuclear and cytoplasmic regions.

Animals↗