scholarly journals A Cellular Reaction to Antibody in Tissue Culture Studied with Electron Microscopy

1959 ◽  
Vol 5 (3) ◽  
pp. 405-410 ◽  
Author(s):  
Harrison Latta
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Endoplasmic Reticulum ◽  
Nuclear Membrane ◽  
Normal Serum ◽  
Butyl Methacrylate ◽  
Heart Cells ◽  
Plasma Clot ◽  
Outer Nuclear Membrane ◽  
Cell Components

The reaction of embryonic chick heart cells grown in tissue culture to specific guinea pig antiserum has been studied with electron microscopy. Heart fragments from chick embryos were cultured with a plasma clot. After being tested with antiserum or normal serum, they were fixed with buffered osmium tetroxide and embedded in butyl methacrylate before removal from the glass culture chamber. Thin cells found by phase microscopy to have reacted were sectioned in a plane parallel to the glass surface on which they had grown. The results confirm and extend observations made previously while the reactions were occurring. The plasma membrane, like that of the red cell, becomes disrupted or less resistant to trauma following the action of antiserum. The membranes of mitochondria and endoplasmic reticulum vesiculate and swell. Before nuclear shrinkage becomes prominent, the outer nuclear membrane separates over a large portion of the nuclear envelope and forms one or more large swollen blebs. Thus, the outer nuclear membrane shows a reactivity similar to endoplasmic reticulum. It is suggested that the various physical and chemical changes observed to follow the action of antibody and complement on fibroblasts may be explained by osmotic pressure differences between various cell components. Some basic similarities to the action of hemolytic agents on red cells are noted.

scholarly journals Endoplasmic reticulum-to-Golgi transitions upon herpes virus infection

F1000Research ◽  
2018 ◽  
Vol 6 ◽  
pp. 1804 ◽  
Author(s):  
Peter Wild ◽  
Andres Kaech ◽  
Elisabeth M. Schraner ◽  
Ladina Walser ◽  
Mathias Ackermann
Keyword(s):  
Endoplasmic Reticulum ◽  
Nuclear Envelope ◽  
Golgi Complex ◽  
Nuclear Membrane ◽  
Progeny Virus ◽  
Cytoplasmic Matrix ◽  
Outer Nuclear Membrane ◽  
Nuclear Membranes ◽  
Perinuclear Space ◽  
Hsv 1

Background: Herpesvirus capsids are assembled in the nucleus, translocated to the perinuclear space by budding, acquiring tegument and envelope, or released to the cytoplasm via impaired nuclear envelope. One model proposes that envelopment, “de-envelopment” and “re-envelopment” is essential for production of infectious virus. Glycoproteins gB/gH were reported to be essential for de-envelopment, by fusion of the “primary” envelope with the outer nuclear membrane. Yet, a high proportion of enveloped virions generated from genomes with deleted gB/gH were found in the cytoplasm and extracellular space, suggesting the existence of alternative exit routes.Methods: We investigated the relatedness between the nuclear envelope and membranes of the endoplasmic reticulum and Golgi complex, in cells infected with either herpes simplex virus 1 (HSV-1) or a Us3 deletion mutant thereof, or with bovine herpesvirus 1 (BoHV-1) by transmission and scanning electron microscopy, employing freezing technique protocols.Results:  The Golgi complex is a compact entity in a juxtanuclear position covered by a membrane on thecisface. Golgi membranes merge with membranes of the endoplasmic reticulum forming an entity with the perinuclear space. All compartments contained enveloped virions. After treatment with brefeldin A, HSV-1 virions aggregated in the perinuclear space and endoplasmic reticulum, while infectious progeny virus was still produced.Conclusions: The data suggest that virions derived by budding at nuclear membranes are intraluminally transported from the perinuclear space via Golgi -endoplasmic reticulum transitions into Golgi cisternae for packaging. Virions derived by budding at nuclear membranes are infective like Us3 deletion mutants, which  accumulate in the perinuclear space. Therefore, i) de-envelopment followed by re-envelopment is not essential for production of infective progeny virus, ii) the process taking place at the outer nuclear membrane is budding not fusion, and iii) naked capsids gain access to the cytoplasmic matrix via impaired nuclear envelope as reported earlier.

Extrusion of Nuclear Material During Oogenesis in Blatta Orientalis

1962 ◽  
Vol s3-103 (62) ◽  
pp. 141-145
Author(s):  
R.A. R. GRESSON ◽  
L. T. THREADGOLD
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Nuclear Membrane ◽  
Close Association ◽  
Nuclear Material ◽  
Blatta Orientalis ◽  
Nucleolar Material

That nucleolar material is extruded to the cytoplasm of the young oocyte of Blatta orientalis is confirmed by means of electron microscopy. The nucleolus and nucleolar extrusions are shown to contain RNA. In addition to the nucleolar extrusions, vesicle-like structures originate in the nuclear membrane and from there pass into the cytoplasm where they become indistinguishable from elements of the endoplasmic reticulum. When the nucleolar extrusions reach the cytoplasm they increase in size, come into close association with a few mitochondria, and migrate towards the periphery of the cell. It is concluded that the emission of material from the nucleolus and the passage of vesicles from the nuclear membrane to the cytoplasm are necessary prerequisites for the process of vitellogenesis.

Mechanism regulating nuclear calcium signalingThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.

2006 ◽  
Vol 84 (3-4) ◽  
pp. 403-422 ◽  
Author(s):  
Anant N. Malviya ◽  
Christian Klein
Keyword(s):  
Endoplasmic Reticulum ◽  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Calcium Signaling ◽  
Nuclear Membrane ◽  
Protein Fragment ◽  
Outer Nuclear Membrane ◽  
Nuclear Calcium ◽  
A Cell ◽  
Epidermal Growth

Although the outer nuclear membrane is continuous with the endoplasmic reticulum, it is possible to isolate nuclei both intact and free from endoplasmic reticulum contaminants. The outer and the inner nuclear membranes can be purified free from cross-contamination. Evidence in support of autonomous regulation of nuclear calcium signaling relies upon the investigations with isolated nuclei. Mechanisms for generating calcium signaling in the nucleus have been identified. Two calcium transporting systems, an ATP-dependant nuclear Ca2+-ATPase and an IP4-mediated inositol 1,3,4,5-tetrakisphosphate receptor, are located on the outer nuclear membrane. Thus, ATP and IP4, depending on external free calcium concentrations, are responsible for filling the nuclear envelope calcium pool. The inositol 1,4,5-trisphosphate receptor is located on the inner nuclear membrane with its ligand binding domain facing toward the nucleoplasm. Likewise, the ryanodine receptor is located on the inner nuclear membrane and its ligand cADP-ribose is generated within the nucleus. A 120 kDa protein fragment of nuclear PLC-γ1 is stimulated in vivo by epidermal growth factor nuclear signaling coincident with the time course of nuclear membrane epidermal growth factor receptor activation. Stimulated 120 kDa protein fragment interacts with PIKE, a nuclear GTPase, and together they form a complex with PI[3]kinase serving as a module for nuclear PI[3]K stimulation. Thus, the nucleus has its own IP3 generating system.

Structure of Cultured Fibroblasts from Dermatosparaxic Calves

1975 ◽  
Vol 12 (1) ◽  
pp. 16-31 ◽  
Author(s):  
J. A. Yaeger ◽  
R. L. Church ◽  
M. L. Tanzer
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Endoplasmic Reticulum ◽  
Cell Lines ◽  
Lipid Droplets ◽  
Cultured Fibroblasts ◽  
Fibrillar Material ◽  
Golgi Region ◽  
Clonal Cell Lines ◽  
Extracellular Fibrils

Clonal cell lines from the dermis of a dermatosparaxic calf were grown in tissue culture. After fixation in a mixture of glutaraldehyde and osmium, they were prepared for electron microscopy. Most cells contained a well-developed Golgi region, lysosomes, mitochondria, and dilated cisternae of rough endoplasmic reticulum. They also contained numerous, large bundles of intracellular filaments, many lipid droplets and extensive arrays of vesicles. Cultures accumulated substantial amounts of extracellular fibrillar material. The fibrils were loosely packed and indistinctly cross-banded. Bundles of intracellular filaments were commonly parallel in adjacent cells and also parallel to extracellular fibrils. These cytoplasmic features may result from the inability of the secreted collagen to form normal fibrils.

Particles Seen in Fefeze-Fracture Preparattoks

1969 ◽  
Vol 27 ◽  
pp. 206-207
Author(s):  
Stanley Bullivant
Keyword(s):  
Endoplasmic Reticulum ◽  
Cardiac Muscle ◽  
Nuclear Membrane ◽  
Actin Filaments ◽  
Carbon Coating ◽  
Freeze Fracture ◽  
Animal Tissues ◽  
Outer Nuclear Membrane ◽  
First Category ◽  
Electron Microscopical

A variety of animal tissues were examined by the simple freeze-fracture technique. An apparatus was used in which the Pt shadowing and carbon coating was carried out through aligned tunnels or apertures. Tn this way the previously fractured cold specimen is protected from contamination.Some particles seen in freeze-fracture replicas can be correlated with structures seen by other electron microscopical techniques. Into this first category fall the broken-off ends of myosin and actin filaments, (Figure 1), the broken-off ends of nucleohistone fibrils the particles seen between membranes in the close junction in cardiac muscle (Figure 2) and quantasomes in chloroplasts. The second group of particles are those found on membranes. An example of these is provided by the membrane faces seen in a fracture through the endoplasmic reticulum (Figure 3). The topology of the situation can be worked out using the outer nuclear membrane as a reference and counting off alternate cytoplasmic and cisternal spaces.

scholarly journals FINE STRUCTURE OF NERVE FIBERS AND GROWTH CONES OF ISOLATED SYMPATHETIC NEURONS IN CULTURE

1973 ◽  
Vol 56 (3) ◽  
pp. 713-735 ◽  
Author(s):  
Mary Bartlett Bunge
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Endoplasmic Reticulum ◽  
Fine Structure ◽  
Sensory Neuron ◽  
Sympathetic Neurons ◽  
Growth Cones ◽  
Nerve Fibers ◽  
Cervical Ganglia ◽  
Neuron Growth

The leading tips of elongating nerve fibers are enlarged into "growth cones" which are seen in tissue culture to continually undergo changes in conformation and to foster numerous transitory slender extensions (filopodia) and/or a veillike ruffling sheet. After explantation of 1-day-old rat superior cervical ganglia (as pieces or as individual neurons), nerve fibers and tips were photographed during growth and through the initial stages of aldehyde fixation and then relocated after embedding in plastic. Electron microscopy of serially sectioned tips revealed the following. The moving parts of the cone, the peripheral flange and filopodia, contained a distinctive apparently filamentous feltwork from which all organelles except membranous structures were excluded; microtubules were notably absent from these areas. The cone interior contained varied forms of agranular endoplasmic reticulum, vacuoles, vesicles, coated vesicles, mitochondria, microtubules, and occasional neurofilaments and polysomes. Dense-cored vesicles and lysosomal structures were also present and appeared to be formed locally, at least in part from reticulum. The possible roles of the various forms of agranular membranous components are discussed and it is suggested that structures involved in both the assembly and degradation of membrane are present in the cone. The content of these growing tips resembles that in sensory neuron growth cones studied by others.

scholarly journals The eta isoform of protein kinase C is localized on rough endoplasmic reticulum.

1994 ◽  
Vol 14 (6) ◽  
pp. 3782-3790 ◽  
Author(s):  
K Chida ◽  
H Sagara ◽  
Y Suzuki ◽  
A Murakami ◽  
S Osada ◽  
...  
Keyword(s):  
Endoplasmic Reticulum ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Rough Endoplasmic Reticulum ◽  
Nuclear Membrane ◽  
Mouse Skin ◽  
Endoplasmic Reticulum Membrane ◽  
Present Observation ◽  
Outer Nuclear Membrane ◽  
Kinase C

The eta isoform of protein kinase C, isolated from a cDNA library of mouse skin, has unique tissue and cellular distributions. It is predominantly expressed in epithelia of the skin, digestive tract, and respiratory tract in close association with epithelial differentiation. We report here that this isoform is localized on the rough endoplasmic reticulum in transiently expressing COS1 cells and constitutively expressing keratinocytes. By the use of polyclonal antibodies raised against peptides of the diverse D1 and D2/D3 regions, we found that immunofluorescent signals were strongest in the cytoplasm around the nucleus and became weaker toward the peripheral cytoplasm. Under immunoelectron microscopic examination, electron-dense signals were located on the rough endoplasmic reticulum and on the outer nuclear membrane which is continuous with the endoplasmic reticulum membrane. However, no signals were detected in the nucleus, inner nuclear membrane, smooth endoplasmic reticulum, Golgi apparatus, mitochondria, or plasma membrane. Treatment of the cells in situ with detergents suggested association of the isoform of protein kinase C with intracellular structures. By immunoblotting, a distinct single band with an M(r) of 80,000 was detected in whole-cell lysate and in rough microsomal and crude nuclear fractions, all of which contain outer nuclear membrane and/or rough endoplasmic reticulum. We further demonstrated the absence of a nuclear localization signal in the pseudosubstrate sequence. The present observation is not consistent with the report of Greif et al. (H. Greif, J. Ben-Chaim, T. Shimon, E. Bechor, H. Eldar, and E. Livneh, Mol. Cell. Biol. 12:1304-1311, 1992).

scholarly journals Biogenesis of the crystalloid endoplasmic reticulum in UT-1 cells: evidence that newly formed endoplasmic reticulum emerges from the nuclear envelope.

1986 ◽  
Vol 102 (6) ◽  
pp. 2158-2168 ◽  
Author(s):  
R K Pathak ◽  
K L Luskey ◽  
R G Anderson
Keyword(s):  
Endoplasmic Reticulum ◽  
Nuclear Envelope ◽  
Nuclear Membrane ◽  
Cholesterol Biosynthesis ◽  
Integral Membrane Protein ◽  
Enzyme Synthesis ◽  
Hmg Coa Reductase ◽  
Outer Nuclear Membrane ◽  
Sequence Of Events ◽  
Coa Reductase

The crystalloid endoplasmic reticulum (ER), a specialized smooth ER of the compactin-resistant UT-1 cell, is composed of multiple membrane tubules packed together in a hexagonal pattern. This membrane contains large amounts of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, an integral membrane protein that enzymatically regulates endogenous cholesterol biosynthesis. Using morphological and immunocytochemical techniques, we have traced the sequence of events in the biogenesis of this ER when compactin-withdrawn UT-1 cells, which do not have a crystalloid ER, are incubated in the presence of compactin. After 15 h of incubation in the presence of compactin, many cells had profiles of ER cisternae that were juxtaposed to the nuclear envelope and studded with ribosomes on their outer membrane. Both the outer nuclear membrane and the ER membrane contained HMG CoA reductase; however, there was little or no detectable enzyme in rough ER that was free in the cytoplasm. With longer times of incubation in the presence of compactin, these cells had lamellar stacks of smooth ER next to the nuclear envelope that contained HMG CoA reductase. Coordinate with the appearance of the smooth ER, crystalloid ER appeared in the same cell. Often regions of continuity were found between the membrane of the smooth ER and the membrane of the crystalloid ER tubules. These studies suggest that HMG CoA reductase is synthesized along the outer nuclear membrane and in response to increased enzyme synthesis, a membrane emerges from the outer nuclear membrane as smooth ER cisternae, which then transforms into crystalloid ER tubules.

scholarly journals ELECTRON MICROSCOPE STUDY OF MAMMALIAN CARDIAC MUSCLE CELLS

1957 ◽  
Vol 3 (2) ◽  
pp. 261-268 ◽  
Author(s):  
Dan H. Moore ◽  
Helmut Ruska
Keyword(s):  
Endoplasmic Reticulum ◽  
Nuclear Membrane ◽  
Heart Muscle ◽  
Plasma Membranes ◽  
Cardiac Muscle Cells ◽  
Outer Nuclear Membrane ◽  
Intercalated Discs ◽  
Cell Plasma ◽  
Entire Cell ◽  
Helicoidal Structure

The cellular theory of heart muscle is supported by a detailed description of the intercalary discs. The discs are adjacent plasma membranes separated by an interspace while the sarcolemma appears as plasma membrane, interspace plus basement membrane of the interstitium. The nucleus of the cell is closely associated with the entire cell by way of the endoplasmic reticulum. Transversely it connects the outer nuclear membrane at the level of the Z and M bands with the contractile material and the sarcolemma. Longitudinally it connects the outer nuclear membrane with the plasmalemma at the intercalated discs. The description of the spiral attachment of the endoplasmic reticulum on the outer nuclear membrane supplements earlier observations on the helicoidal structure of the heart muscle cell. Plasma membranes and endoplasmic reticulum are considered to be carriers of membrane potentials and to conduct excitation.

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