Origine et formation de différents types de vacuoles induites par la multiplication d'Alphavirus Sindbis dans divers systemes cellulaires

1979 ◽  
Vol 25 (12) ◽  
pp. 1452-1459 ◽  
Author(s):  
Yves Lombard ◽  
Philippe Poindron ◽  
Aimé Porte
Keyword(s):  
Endoplasmic Reticulum ◽  
Glial Cells ◽  
Chicken Embryo ◽  
Cytoplasmic Membrane ◽  
Dense Material ◽  
Newborn Mice ◽  
Electron Dense Material ◽  
Double Membrane ◽  
Single Membrane ◽  
Chicken Embryo Fibroblasts

Spherule-containing vacuoles and nucleocapsid-bearing vacuoles (cytopathic vacuoles types 1 and 2 respectively of Grimley et al. 1968) induced by Alphavirus Sindbis were studied in brains from newborn mice, chicken embryo fibroblasts, and two lines of tumoral glial cells from muridae. Endoplasmic reticulum (ER) elements and finely granular electron-dense material also seen in contact with nucleocapsids seemed to be involved in the formation of the classical single-membrane spherule-containing vacuoles. A second type of spherule-containing vacuoles were characterized by their double membrane and an amorphous electron-dense content and were probably derived from mitochondria. Nucleocapsid-bearing vacuoles were formed from modified ER elements and seemed to be linked to excessive synthesis of viral material. Such ER alterations were not observed in RG6 cells. In these cells, there were only spherule-containing vacuoles, while nucleocapsids were seen associated with the cytoplasmic membrane only.

scholarly journals The Origin, Dynamic Morphology, and PI4P-Independent Formation of Encephalomyocarditis Virus Replication Organelles

mBio ◽  
2018 ◽  
Vol 9 (2) ◽  
pp. e00420-18 ◽  
Author(s):  
C. E. Melia ◽  
H. M. van der Schaar ◽  
A. W. M. de Jong ◽  
H. R. Lyoo ◽  
E. J. Snijder ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Rna Synthesis ◽  
Electron Tomography ◽  
Membrane Vesicles ◽  
Encephalomyocarditis Virus ◽  
Escape Mutant ◽  
Membrane Structures ◽  
Double Membrane ◽  
Single Membrane ◽  
Phosphatidylinositol 4

ABSTRACTPicornaviruses induce dramatic rearrangements of endomembranes in the cells that they infect to produce dedicated platforms for viral replication. These structures, termed replication organelles (ROs), have been well characterized for theEnterovirusgenus of thePicornaviridae. However, it is unknown whether the diverse RO morphologies associated with enterovirus infection are conserved among other picornaviruses. Here, we use serial electron tomography at different stages of infection to assess the three-dimensional architecture of ROs induced by encephalomyocarditis virus (EMCV), a member of theCardiovirusgenus of the family of picornaviruses that is distantly related. Ultrastructural analyses revealed connections between early single-membrane EMCV ROs and the endoplasmic reticulum (ER), establishing the ER as a likely donor organelle for their formation. These early single-membrane ROs appear to transform into double-membrane vesicles (DMVs) as infection progresses. Both single- and double-membrane structures were found to support viral RNA synthesis, and progeny viruses accumulated in close proximity, suggesting a spatial association between RNA synthesis and virus assembly. Further, we explored the role of phosphatidylinositol 4-phosphate (PI4P), a critical host factor for both enterovirus and cardiovirus replication that has been recently found to expedite enterovirus RO formation rather than being strictly required. By exploiting an EMCV escape mutant, we found that low-PI4P conditions could also be overcome for the formation of cardiovirus ROs. Collectively, our data show that despite differences in the membrane source, there are striking similarities in the biogenesis, morphology, and transformation of cardiovirus and enterovirus ROs, which may well extend to other picornaviruses.IMPORTANCELike all positive-sense RNA viruses, picornaviruses induce the rearrangement of host cell membranes to form unique structures, or replication organelles (ROs), that support viral RNA synthesis. Here, we investigate the architecture and biogenesis of cardiovirus ROs and compare them with those induced by enteroviruses, members of the well-characterized picornavirus genusEnterovirus. The origins and dynamic morphologies of cardiovirus ROs are revealed using electron tomography, which points to the endoplasmic reticulum as the donor organelle usurped to produce single-membrane tubules and vesicles that transform into double-membrane vesicles. We show that PI4P, a critical lipid for cardiovirus and enterovirus replication, is not strictly required for the formation of cardiovirus ROs, as functional ROs with typical morphologies are formed under phosphatidylinositol 4-kinase type III alpha (PI4KA) inhibition in cells infected with an escape mutant. Our data show that the transformation from single-membrane structures to double-membrane vesicles is a conserved feature of cardiovirus and enterovirus infections that likely extends to other picornavirus genera.

scholarly journals Calcium-containing structures in vertebrate glial cells. Ultrastructural and microprobe analysis.

1975 ◽  
Vol 64 (2) ◽  
pp. 322-330 ◽  
Author(s):  
P Gambetti ◽  
S E Erulkar ◽  
A P Somlyo ◽  
N K Gonatas
Keyword(s):  
Glial Cells ◽  
Electron Probe Microanalysis ◽  
Electron Probe ◽  
Microprobe Analysis ◽  
Dense Material ◽  
Electron Dense Material ◽  
Human Astrocytes ◽  
Calcium And Phosphorus ◽  
Osmiophilic Particles

Electron probe microanalysis has revealed that vesicular or cisternal structures containing electron-dense material in frog ependymal glial cells contain deposits of calcium and phosphorus. The so-called "osmiophilic particles" in human astrocytes also contain calcium. It is suggested that these organelles are storage sites of calcium.

The transformation of muscle into bioluminescent tissue in the fish Benthalbella infans Zugmayer

1985 ◽  
Vol 225 (1239) ◽  
pp. 213-218 ◽  
Keyword(s):  
Lattice Structure ◽  
Dense Material ◽  
Muscle Fibres ◽  
Normal Muscle ◽  
Electron Dense Material ◽  
Double Membrane ◽  
Dense Granules ◽  
Sarcotubular System ◽  
Luminescent System ◽  
Distinct Membrane

The luminescent tissue of the pelvic and anal organs of the scopelarchid fish Benthalbella infans Zugmayer is derived from skeletal muscle. Normal muscle cells are found to be interspersed between fibres containing ‘granules’ 0.5-2 µm in diameter. The majority of granules are bounded by a double membrane and filled with an electron-dense material which is presumably a component of the luminescent system. All fibres with granules contain remnants of myofibrils, although there is a wide variation in their ultrastructure. In some cases, relatively few granules are present and the myofilament lattice structure and the sarcotubular system of myofibrils appears normal. Other fibres are almost entirely filled with granules and only isolated fragments of degenerating myofibrils and free filaments are present. In these fibres, there are relatively few mitochondria and glycogen granules are more numerous than in normal muscle fibres. Some of the dense granules are not bounded by a distinct membrane; they have a diffuse periphery and ‘empty’ luminescent vesicles containing small amounts of electron-dense material are always present. Lysosomes are common to all muscle fibres containing granules and frequently enclose complete granules, suggesting their involvement in the turnover or processing of luminescent material. The modification of muscle fibres for other physiological purposes, and the origin of luminous tissue in scopelarchid fish are briefly discussed.

The ultrastructure of the R.S. zoospore of the Chytridiomycete Physoderma gerhardti

1978 ◽  
Vol 56 (12) ◽  
pp. 1387-1393 ◽  
Author(s):  
Robert J. Lowry ◽  
Frederick K. Sparrow
Keyword(s):  
Endoplasmic Reticulum ◽  
Fine Structure ◽  
Inclusion Bodies ◽  
Smooth Endoplasmic Reticulum ◽  
Lipid Body ◽  
Dense Material ◽  
Electron Dense Material ◽  
General Internal ◽  
Deep Groove ◽  
Posterior Flagellum

The fine structure of the zoospore of Physoderma gerhardti Schroeter is described. It possesses a single very large lipid body (sometimes accompanied by several smaller ones) situated laterally to the nuclear cap – nucleus complex and is associated with electron-dense material (the microbody) and the single large posteriorly located mitochondrion. The single posterior flagellum proximally terminates in a kinetosome just short of the posterior cone-shaped end of the nucleus. The kinetosome lies in a deep groove in the mitochondrion and is associated with this organelle by striated rootlets. The kinetosome terminates in electron-dense material from which an array of microtubules arises. These microtubules run along the sides of the cone-shaped nucleus and nuclear cap. There is an accessory centriole lying close to and more or less parallel with the kinetosome. The cytoplasm contains a small amount of smooth endoplasmic reticulum and several inclusion bodies in the anterior region and several small vacuoles in the posterior region of the cell. The general internal organization of the zoospore of Physoderma gerhardti more nearly resembles that of a blastocladiaceous fungus than of any chytrid thus far investigated.

scholarly journals Fibrillogenesis in the wax-moth, Galleria mellonella

1964 ◽  
Vol s3-105 (72) ◽  
pp. 391-403
Author(s):  
DOREEN E. ASHHURST
Keyword(s):  
Endoplasmic Reticulum ◽  
Connective Tissue ◽  
Plasma Membranes ◽  
Galleria Mellonella ◽  
Fibrous Tissue ◽  
Dense Material ◽  
Fibrous Connective Tissue ◽  
Electron Dense Material ◽  
Wax Moth ◽  
Great Development

The fibroblasts of the pupa are characterized by the great development of the endoplasmic reticulum, which becomes dilated to form vesicles containing a rather electron-dense material which is thought to be a precursor of the collagen fibrils. Fibrils are seen within the cytoplasm of the fibroblasts; these are about 12.5 to 20 mµ. in diameter and some ofthem show indications of banding with a periodicity between 1 5 and 20 mµ. It is thought that these fibrils and their surrounding cytoplasm become incorporated into the fibrous connective tissue. The plasma membranes of the fibroblasts are discontinuous where they are adjacent to the fibrous tissue. The fibrils in the connective tissue are obscured by masses of mucopolysaccharide, but there are indications that these fibrils are essentially similar to the intracellular fibrils. The processes of fibrillogenesis in the moth and in various vertebrate tissues are shown to have many features in common.

Comparative Morphology of Intercellular Bridges Between Developing Germ Cells of the Ovary

1970 ◽  
Vol 28 ◽  
pp. 328-329
Author(s):  
J. R. Ruby ◽  
R. F. Dyer ◽  
R. G. Skalko ◽  
R. F. Gasser ◽  
E. P. Volpe
Keyword(s):  
Germ Cells ◽  
Rana Pipiens ◽  
Comparative Morphology ◽  
Dense Material ◽  
Individual Species ◽  
Microscope Examination ◽  
Electron Dense Material ◽  
Intercellular Bridges ◽  
Cytoplasmic Side ◽  
Intercellular Connections

An electron microscope examination of fetal ovaries has revealed that developing germ cells are connected by intercellular bridges. In this investigation several species have been studied including human, mouse, chicken, and tadpole (Rana pipiens). These studies demonstrate that intercellular connections are similar in morphology regardless of the species.Basically, all bridges are characterized by a band of electron-dense material on the cytoplasmic side of the tri-laminar membrane surrounding the connection (Fig.l). This membrane is continuous with the plasma membrane of the conjoined cells. The dense material, however, never extends beyond the limits of the bridge. Variations in the configuration of intercellular connections were noted in all ovaries studied. However, the bridges in each individual species usually exhibits one structural characteristic seldom found in the others. For example, bridges in the human ovary very often have large blebs projecting from the lateral borders whereas the sides of the connections in the mouse gonad merely demonstrate a slight convexity.

The ultrastructure of the double membrane-bound bodies and endoplasmic reticulum in serial sections of wheat spot mosaic-affected wheat plants

1990 ◽  
Vol 48 (3) ◽  
pp. 694-695
Author(s):  
M. H. Chen ◽  
C. Hiruki
Keyword(s):  
Endoplasmic Reticulum ◽  
High Resolution ◽  
Membrane Structure ◽  
Mosaic Disease ◽  
Serial Sections ◽  
Thin Sections ◽  
Double Membrane ◽  
Southern Alberta ◽  
Membrane Bound ◽  
Scanning Em

Wheat spot mosaic disease was first discovered in southern Alberta, Canada, in 1956. A hitherto unidentified disease-causing agent, transmitted by the eriophyid mite, caused chlorosis, stunting and finally severe necrosis resulting in the death of the affected plants. Double membrane-bound bodies (DMBB), 0.1-0.2 μm in diameter were found to be associated with the disease.Young tissues of leaf and root from 4-wk-old infected wheat plants were fixed, dehydrated, and embedded in Spurr’s resin. Serial sections were collected on slot copper grids and stained. The thin sections were then examined with a Hitachi H-7000 TEM at 75 kV. The membrane structure of the DMBBs was studied by numbering them individually and tracing along the sections to see any physical connection with endoplasmic reticulum (ER) membranes. For high resolution scanning EM, a modification of Tanaka’s method was used. The specimens were examined with a Hitachi Model S-570 SEM in its high resolution mode at 20 kV.

scholarly journals Tropomyosin isoforms in chicken embryo fibroblasts: purification, characterization, and changes in Rous sarcoma virus-transformed cells.

1985 ◽  
Vol 100 (3) ◽  
pp. 692-703 ◽  
Author(s):  
J J Lin ◽  
D M Helfman ◽  
S H Hughes ◽  
C S Chou
Keyword(s):  
Rous Sarcoma Virus ◽  
Chicken Embryo ◽  
Actin Binding ◽  
Transformed Cells ◽  
Two Dimensional ◽  
Tropomyosin Isoforms ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Embryo Fibroblasts ◽  
Chicken Embryo Fibroblasts

Seven polypeptides (a, b, c, 1, 2, 3a, and 3b) have been previously identified as tropomyosin isoforms in chicken embryo fibroblasts (CEF) (Lin, J. J.-C., Matsumura, F., and Yamashiro-Matsumura, S., 1984, J. Cell. Biol., 98:116-127). Spots a and c had identical mobility on two-dimensional gels with the slow-migrating and fast-migrating components, respectively, of chicken gizzard tropomyosin. However, the remaining isoforms of CEF tropomyosin were distinct from chicken skeletal and cardiac tropomyosins on two-dimensional gels. The mixture of CEF tropomyosin has been isolated by the combination of Triton/glycerol extraction of monolayer cells, heat treatment, and ammonium sulfate fractionation. The yield of tropomyosin was estimated to be 1.4% of total CEF proteins. The identical set of tropomyosin isoforms could be found in the antitropomyosin immunoprecipitates after the cell-free translation products of total poly(A)+ RNAs isolated from CEF cells. This suggested that at least seven mRNAs coding for these tropomyosin isoforms existed in the cell. Purified tropomyosins (particularly 1, 2, and 3) showed different actin-binding abilities in the presence of 100 mM KCl and no divalent cation. Under this condition, the binding of tropomyosin 3 (3a + 3b) to actin filaments was significantly weaker than that of tropomyosin 1 or 2. CEF tropomyosin 1, and probably 3, could be cross-linked to form homodimers by treatment with 5,5'-dithiobis-(2-nitrobenzoate), whereas tropomyosin a and c formed a heterodimer. These dimer species may reflect the in vivo assembly of tropomyosin isoforms, since dimer formation occurred not only with purified tropomyosin but also with microfilament-associated tropomyosin. The expression of these tropomyosin isoforms in Rous sarcoma virus-transformed CEF cells has also been investigated. In agreement with the previous report by Hendricks and Weintraub (Proc. Natl. Acad. Sci. USA., 78:5633-5637), we found that major tropomyosin 1 was greatly reduced in transformed cells. We have also found that the relative amounts of tropomyosin 3a and 3b were increased in both the total cell lysate and the microfilament fraction of transformed cells. Because of the different actin-binding properties observed for CEF tropomyosins, changes in the expression of these isoforms may, in part, be responsible for the reduction of actin cables and the alteration of cell shape found in transformed cells.

The ultrastructure of M1-a-mediated resistance to powdery mildew infection in barley

1975 ◽  
Vol 53 (22) ◽  
pp. 2589-2597 ◽  
Author(s):  
H. H. Edwards
Keyword(s):  
Powdery Mildew ◽  
Host Cell ◽  
Electron Density ◽  
Epidermal Cells ◽  
Dense Material ◽  
Ultrastructural Changes ◽  
Electron Dense Material ◽  
Host Cytoplasm ◽  
Powdery Mildew Infection ◽  
Cell Protoplast

M1-a-mediated resistance in barley to invasion by the CR3 race of Erysiphe graminis f. sp. hordei does not occur in every host cell with the same speed and severity. In some cells ultrastructural changes within the host cell as a result of resistance will occur within 24 h after inoculation, whereas in other cells these changes may take up to 72 h. In some cells the ultrastructural changes are so drastic that they give the appearance of a hypersensitive death of the host cell, whereas in other cells the changes are very slight. In any case, at the end of these changes the fungus ceases growth. The ultrastructural changes occur in penetrated host epidermal cells as well as non-infected adjacent epidermal and mesophyll cells.The following ultrastructural changes have been observed: (1) an electron-dense material which occurs either free in the vacuole or adhering to the tonoplast (the material is granular or in large clumps); (2) an increased electron density of the host cytoplasm and nucleus; (3) a breakdown of the tonoplast so that the cytoplasmic constituents become dispersed throughout the cell lumen; and (4) the deposition of papillar-like material in areas other than the penetration site. The first three changes take place within the host cell protoplasts and are directly attributable to the gene M1-a. These changes are typical of stress or incompatibility responses and thus M1-a appears to trigger a generalized incompatibility response in the presence of race CR3. The papillar-like material occurs outside the host cell protoplast in the same manner as the papilla and probably is not directly attributable to M1-a.

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