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M A EPSTEIN

Publications and source records attributed to M A EPSTEIN.

At least 37 records · Page 2Linked to original sources
PubMed1963

THE LOCALIZATION BY ELECTRON MICROSCOPY OF HELA CELL SURFACE ENZYMES SPLITTING ADENOSINE TRIPHOSPHATE.

Cultures of normally proliferating Hela cells have been examined in thin sections by electron microscopy following glutaraldehyde fixation, staining in Wachstein and Meisel's adenosine triphosphate containing medium, postosmication, and embedding in an epoxy resin. The cells were stained in suspension in order to ensure uniform accessibility to reagents. Discrete localization of the enzyme reaction product (lead phosphate) was found at the plasma membranes of about half the cells, but nowhere else. It appeared in the form of an intensely electron-opaque deposit lying close against the outer surface of the cells and varying in amount from a chain of small particles to a dense band about 30 mmicro in width. This opaque reaction product was present over microvilli when absent elsewhere on a cell, was heaviest where microvilli and processes were profuse, and was minimal or lacking where cell surfaces were smooth. These observations are discussed in relation to both the idea that surface enzyme activity varies with the physiological phase of individual cells in a population, and the problem of how such enzyme activity becomes manifest at a given site on a morphologically changing membrane system.

Adenosine Triphosphatases↗
PubMed1963

ELECTRON MICROSCOPE OBSERVATIONS ON THE SURFACE ADENOSINE TRIPHOSPHATASE-LIKE ENZYMES OF HELA CELLS INFECTED WITH HERPES VIRUS.

HeLa cells infected with herpes simplex virus have been examined in thin sections by electron microscopy after cytochemical staining for the presence of surface enzymes splitting adenosine triphosphate. As with uninfected HeLa cultures (18), the opaque enzyme reaction product was localized at the plasma membranes of about half the cells, tending to be present where there were microvilli and absent on smooth surfaces. Where mature extracellular herpes particles were found in association with cell membranes showing the enzyme activity, they were invariably likewise stained, and conversely, those mature particles which lay close against cells without reaction product at the surface were themselves free of it. Particles found budding into cytoplasmic vacuoles were also always without opaque deposit since this was never seen at vacuolar membranes, even in cells having the activity at the surface. The enzyme reaction product thus provided a marker indicating the manner in which the particles escape from cells and mature by budding out through cellular membranes, carrying, in the process, a portion of the latter on to themselves to form the outer viral limiting membrane. In some instances, virus particles were observed with more opaque material covering them than was present at the cell membrane with which they were associated. This finding has been taken as evidence for a physiological waxing and waning of surface enzyme activity of adenosine triphosphatase type. The fine structure of the mature extracellular virus as prepared here, using glutaraldehyde fixation, is also recorded. The observations and interpretations are discussed in full.

Adenosine Triphosphatases↗
PubMed1962

Observations on the fine structure of mature herpes simplex virus and on the composition of its nucleoid.

The fine structure and composition of mature herpes virus have been investigated in thin sections by electron microscopy. The virus was grown in cultured HeLa cells and was collected with them. Tests for the biological activity of the infected cultures were included in the first half of the work. Preparations were fixed with both osmium and permanganate, and were embedded either in methacrylate or in aquon, a water-miscible fraction of a commercially available epoxy resin. In further experiments material fixed with permanganate was subjected to the action of specific nucleases or control medium before embedding. All the preparations showed numerous uniform particles around and between the cells and this was paralleled by considerable biological activity where tests were made on samples of the culture fluids. Mature herpes virus has been found to be round and to measure, when dehydration shrinkage is avoided, about 165 mmicro in diameter. The particle contained an eccentric round-ended rod-shaped, electron-opaque nucleoid lying in an inner zone of low density. A dense outer zone or viroplasm surrounded this, no membrane being present between the two zones. After permanganate fixation the particle was found to have an outer limiting membrane showing a triple-layered structure morphologically indistinguishable from that of the plasma membrane of the HeLa cells. The results of the digestion experiments show that herpes virus contains nucleic acid of deoxyribose type and that this is localized in the dense nucleoid. Both the findings and the methods used are discussed.

Cell Line↗
PubMed1962

Observations on the mode of release of herpes virus from infected HeLa cells.

The development of a well adapted strain of herpes virus has been studied in HeLa cells using thin sectioning techniques for electron microscopy. Particular attention was directed to events in the cytoplasm and certain new features were observed. Profuse immature particles with a nucleoid and single limiting membrane were present in the nuclei of infected cells, often in crystalline array; morphologically indistinguishable immature particles were also found very frequently in the cytoplasm. Cells with such particles were intact and well preserved, and contained smooth vacuoles apparently derived from the Golgi component of the endoplasmic reticulum. The cytoplasmic particles escaped from the cells by bulging out as buds through the cell membrane or through that of the cytoplasmic vacuoles until they were attached only by a pedicle and then became free. During this process the particles were gradually enclosed by the membrane through which they passed and carried a coat of it with them as they matured. After permanganate fixation the triple-layered structure of the cell membrane and vacuolar membranes was evident and was identical with that of the outer coat of the mature virus. These findings are discussed both in relation to different types of virus structure and to function in the endoplasmic reticulum and cell membrane.

Animals↗
PubMed1961

Some unusual features of fine structure observed in HeLa cells.

HeLa cells from conventional culture media have been studied in thin sections with the electron microscope; in many cases cells were examined in sets of sections cut in series. The fine structure of the cells is described including three unusual features not hitherto reported. It has been found that numerous cells contained rows of parallel smooth surfaced cisternae spaced about 150 mmicro apart and communicating with rough surfaced elements of the endoplasmic reticulum. These cisternae resembled "annulate lamellae" but did not contain regular arrays of pores. In many cells an area of juxtanuclear cytoplasm was occupied by a membranous structure composed of closely applied pairs of narrow cisternae either arranged in concentric rings or else extending in several directions in a haphazard manner. Sparse particles were present on the outer membranes of each pair of cisternae. Communications between the double cisternae and other membrane-bounded structures were not observed. A small number of cells contained areas of cytoplasm devoid of organelles and filled with amorphous fuzzy material. The observations recorded are discussed.

Animals↗