scholarly journals Studies of membrane fusion. IV. Fusion of HeLa cells with Sendai virus

1979 ◽  
Vol 36 (1) ◽  
pp. 73-84
Author(s):  
S. Knutton
Keyword(s):  
Plasma Membrane ◽  
Membrane Fusion ◽  
Cell Fusion ◽  
Hela Cells ◽  
Sendai Virus ◽  
Freeze Fracture ◽  
Viral Envelope ◽  
Cell Plasma Membrane ◽  
Virus Particles ◽  
Cell Plasma

The Sendai virus-induced fusion of HeLa cells has been studied by freeze-fracture electron microscopy. Freeze-fracture observations confirm previous scanning electron-microscope studies (1977) and show that at 4 degrees C virus particles bind to the cell surface and that cell agglutination results from the crosslinking by virus particles of microvilli on adjacent cells. Incubation at 37 degrees C initiates a change in viral envelope structure and fusion of ‘altered’ virus particles with the cell plasma membrane. Fusion of a virus particle with two crosslinked cells is probably the membrane fusion event which initiates cell-cell fusion; fusion is completed as a result of virally induced cell swelling. Lateral diffusion of viral envelope components following virus-cell fusion and, in some instances, an aggregation of plasma membrane intramembrane particles occurs in swollen cells. These observations show that the mechanisms of viral envelope-cell and probably cell-cell fusion are the same as have been reported for erythrocytes. Although endocytosis of intact virus particles does occur, the specialized cell-mediated mechanism for fusion of the viral envelope with the cell plasma membrane suggests that this, and not viropexis, is the mechanism of Sendai virus infection.

Studies of membrane fusion. VI. Mechanism of the membrane fusion and cell swelling stages of Sendai virus-mediated cell fusion

1980 ◽  
Vol 43 (1) ◽  
pp. 103-118
Author(s):  
S. Knutton
Keyword(s):  
Membrane Fusion ◽  
Cell Fusion ◽  
Sendai Virus ◽  
Freeze Fracture ◽  
Viral Envelope ◽  
Cell Swelling ◽  
Fusion Event ◽  
Virus Envelope ◽  
Membrane Rupture ◽  
Membrane Tubules

The membrane fusion and cell swelling stages of Sendai virus-mediated cell-cell fusion have been studied by thin-section and freeze-fracture electron microscopy. Sites of membrane fusion have been detected in human erythrocytes arrested at the membrane fusion stage of cell fusion and in virtually all cases a fused viral envelope or envelope components has been identified thus providing further direct evidence that cell-viral envelope-cell bridge formation is the membrane fusion event in Sendai virus-induced cell fusion. Radial expansion of a single virus bridge connecting 2 cells is sufficient to produce a fused cell. Membrane redistribution which occurs during this cell swelling stage of the fusion process is often accompanied by the formation of a system of membrane tubules in the plane of expansion of the virus bridge. The tubules originate from points of fusion between the bridging virus envelope and the erythrocyte membrane and also expand radially as cells swell. Ultimately membrane rupture occurs and the tubules appear to break down as small vesicles. When previously observed in cross-sectioned cells these membrane tubules were interpreted as sites of direct membrane fusion. The present study indicates that this interpretation is incorrect and shows that the tubules are generated subsequent to membrane fusion when 2 cells connected by a virus bridge are induced to swell. A mechanism to explain the formation of this system of membrane tubules is proposed.

Studies of membrane fusion. V. Fusion of erythrocytes with non-haemolytic Sendai virus

1979 ◽  
Vol 36 (1) ◽  
pp. 85-96
Author(s):  
S. Knutton
Keyword(s):  
Membrane Fusion ◽  
Cell Fusion ◽  
Erythrocyte Membrane ◽  
Sendai Virus ◽  
Lateral Diffusion ◽  
Viral Envelope ◽  
Cell Swelling ◽  
Viral Membrane ◽  
Cytoplasmic Bridges ◽  
Viral Envelopes

The fusion of human erythrocytes with non-haemolytic ‘1-day’ Sendai virus has been studied by electron microscopy. The mechanism of viral envelope-cell fusion is the same as that described previously for haemolytic ‘3-day’ Sendai virus except that fusion is frequently arrested at an initial stage when 2 segments of smooth linear viral membrane fuse and become incorporated into the erythrocyte membrane. After longer periods of incubation at 37 degrees C, in addition to many partly fused virus particles, long (up to 4 micrometer) lengths of smooth linear viral membrane are seen within the erythrocyte membrane which arise by linear aggregation of shorter (approximately 0.25 micrometer long) segments of smooth linear membrane derived from individual fused viral envelopes. Cell-Cell fusion, as a result of the fusion of a viral envelope with 2 adjacent erythrocytes also occurs but, in the absence of cell swelling, fusion is arrested at this stage with cells joined by one (or more) small cytoplasmic bridges. Typical fused cells are produced if such cells are swollen with hypotonic buffer. These observations provide further evidence that membrane fusion and cell swelling are distinct events in cell fusion and that cell swelling is the driving force both for completing the incorporation of the viral envelope into the cell membrane and for expanding cells connected by small cytoplasmic bridges to form spherical fused cells. Little lateral diffusion of viral envelope components occurs in the absence of cell swelling; in fact, some aggregation of components occurs. Comparison with previous studies using haemolytic ‘3-day’ Sendai virus suggests that virally induced cell swelling perturbs membrane structure so as to allow the rapid lateral diffusion of integrated viral envelope components.

An Ultrastructural Study of Interspecific Cell Fusion Induced by Inactivated Sendai Virus

1966 ◽  
Vol 1 (4) ◽  
pp. 401-406
Author(s):  
E. E. SCHNEEBERGER ◽  
H. HARRIS
Keyword(s):  
Cell Fusion ◽  
Hela Cells ◽  
Sendai Virus ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Virus Particles ◽  
Large Numbers ◽  
Ehrlich Ascites ◽  
Ehrlich Ascites Cells ◽  
Cytoplasmic Bridges

An electron-microscopic examination was made of the process of cell fusion induced by Sendai virus inactivated by ultraviolet light. Ehrlich ascites cells, HeLa cells, rabbit macrophages, rat lymphocytes and nucleated hen erythrocytes were chosen for study because it had previously been shown that these cells could be fused together, with varying degrees of facility, to form artificial heterokaryons. Cells which had large numbers of microvilli on their surfaces fused together more readily than those which had not, but the presence of microvilli was not essential for fusion to occur. Fusion appeared in all cases to be initiated by the formation of small cytoplasmic bridges between the cells; but virus particles, although present elsewhere on the surface of the cells, were not detected at or near the cytoplasmic bridges. HeLa-hen erythrocyte heterokaryons were formed by the fusion of HeLa cells with red cell ghosts.

scholarly journals Morphological changes of the adipose cell plasma membrane during lipolysis.

1977 ◽  
Vol 72 (1) ◽  
pp. 104-117 ◽  
Author(s):  
J Carpentier ◽  
A Perrelet ◽  
L Orci
Keyword(s):  
Plasma Membrane ◽  
Morphological Changes ◽  
Freeze Fracture ◽  
Total Content ◽  
Structural Features ◽  
Epididymal Adipose Tissue ◽  
Local Concentration ◽  
Cell Plasma Membrane ◽  
Adipose Cell ◽  
Cell Plasma

Morphological changes of the plasma membrane in the white adipose cell associated with lipid mobilization were assessed qualitatively and quantitatively on freeze-fracture replicas of epididymal adipose tissue from fasted and from streptozotocin-diabetic rats. The number of plasma membrane invaginations and intramembranous particles were evaluated per square micrometer of membrane and per entire adipocyte. These two determinations show that the number per square micrometer (local concentration) of both structural features progressively increases with the duration of diabetes and fasting, while that at the same time their number per entire cell (total content) remains unchanged. These data thus show: (a) a reorganization of the adipose cell plasma membrane during lipolysis; and (b) that this reorganization can be detected only by determining the concentration and the total content of the structural features of the membrane involved.

scholarly journals Monitoring of the Zeta Potential of Human Cells upon Reduction in Their Viability and Interaction with Polymers

Acta Naturae ◽  
2012 ◽  
Vol 4 (1) ◽  
pp. 78-81 ◽  
Author(s):  
O. V. Bondar ◽  
D. V. Saifullina ◽  
I. I. Shakhmaeva ◽  
I. I. Mavlyutova ◽  
T. I. Abdullin
Keyword(s):  
Plasma Membrane ◽  
Light Scattering ◽  
Zeta Potential ◽  
Hela Cells ◽  
Human Cells ◽  
Cell Plasma Membrane ◽  
Cell Plasma ◽  
Wide Range ◽  
Different Types ◽  
The Difference

The dynamic light scattering (DLS) technique was applied in order to assess the zeta potential of the plasma membrane of human cells. At pH 7.4, the cell zeta potential for different types of cells showed variations over a wide range and was equal to -19.4 0.8 mV for HeLa cells and -31.8 1.1 mV for erythrocytes. The difference could presumably be attributed to the differences in the biochemical composition of the cell plasma membrane. As a result of the heating of HeLa cells, the zeta potential shifted towards more negative voltages by 4.2 mV. An increase in the zeta potential correlated with an increase in the content of phosphatidylserine on the cell surface, which is considered to be an early marker of apoptosis. The DLS technique was also used to study the interactions between the cells and membranotropic polymers, such as polycations and nonionogenic Pluronic L121.

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