Giant unilamellar vesicle formation technique allows study of membrane-bound biological systems

Viruses are highly dependent on the host they infect. Their dependence triggers processes of virus–host co-adaptation, enabling viruses to explore host resources whilst escaping immunity. Scientists have tackled viral–host interplay at differing levels of complexity—in individual hosts, organs, tissues and cells—and seminal studies advanced our understanding about viral lifecycles, intra- or inter-species transmission, and means to control infections. Recently, it emerged as important to address the physical properties of the materials in biological systems; membrane-bound organelles are only one of many ways to separate molecules from the cellular milieu. By achieving a type of compartmentalization lacking membranes known as biomolecular condensates, biological systems developed alternative mechanisms of controlling reactions. The identification that many biological condensates display liquid properties led to the proposal that liquid–liquid phase separation (LLPS) drives their formation. The concept of LLPS is a paradigm shift in cellular structure and organization. There is an unprecedented momentum to revisit long-standing questions in virology and to explore novel antiviral strategies. In the first part of this review, we focus on the state-of-the-art about biomolecular condensates. In the second part, we capture what is known about RNA virus-phase biology and discuss future perspectives of this emerging field in virology.

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Two Distinct Types of Microfilaments in the Cytoplasm of Human Adenohypophysial Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100073787 ◽

1973 ◽

Vol 31 ◽

pp. 668-669

Author(s):

E. Horvath ◽

K. Kovacs ◽

I. E. Stratmann ◽

C. Ezrin

Keyword(s):

Pituitary Tumors ◽

Average Diameter ◽

Anterior Lobe ◽

Uranyl Acetate ◽

Type I ◽

Breast Cancer Patients ◽

Human Pituitary ◽

Severe Diabetes ◽

Pituitary Glands ◽

Membrane Bound

Surgically removed human pituitary glands as well as pituitary tumors fixed in glutaraldehyde, postfixed in osmium tetroxide, embedded in epon resin, stained with uranyl acetate and lead citrate have been investigated by electron microscopy in order to correlate ultrastructure with functional activity. In the course of this study two distinct types of microfilaments have been identified in the cytoplasm of adenohypophysiocytes.Type I microfilaments (Fig. 1) were found in the cytoplasm of anterior lobe cells of five female subjects with disseminated mammary cancer and two patients with severe diabetes mellitus. The breast cancer patients were treated pre-operatively for various periods of time with different doses of oxysteroids. The microfilaments had an average diameter of JO A, formed parallel bundles, were scattered irregularly in the cytoplasm and were frequently located in the perikaryon. They were not membrane-bound and failed to show any periodicity.

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Transmission Electron Microscopy of Protein Macromolecules

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066176 ◽

1971 ◽

Vol 29 ◽

pp. 424-425

Author(s):

Henry S. Slayter

Keyword(s):

Biological Systems ◽

Metal Vapor ◽

Resolving Power ◽

Electron Microscopic ◽

Chemical Methods ◽

Molecular Weights ◽

The Past ◽

Physical Chemical ◽

Transmission Electron

Electron microscopic methods have been applied increasingly during the past fifteen years, to problems in structural molecular biology. Used in conjunction with physical chemical methods and/or Fourier methods of analysis, they constitute powerful tools for determining sizes, shapes and modes of aggregation of biopolymers with molecular weights greater than 50, 000. However, the application of the e.m. to the determination of very fine structure approaching the limit of instrumental resolving power in biological systems has not been productive, due to various difficulties such as the destructive effects of dehydration, damage to the specimen by the electron beam, and lack of adequate and specific contrast. One of the most satisfactory methods for contrasting individual macromolecules involves the deposition of heavy metal vapor upon the specimen. We have investigated this process, and present here what we believe to be the more important considerations for optimizing it. Results of the application of these methods to several biological systems including muscle proteins, fibrinogen, ribosomes and chromatin will be discussed.

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Intranuclear Crystals in Regenerating Canine Kidneys

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100072502 ◽

1973 ◽

Vol 31 ◽

pp. 412-413

Author(s):

D.G. Osborne ◽

L.J. McCormack ◽

M.O. Magnusson ◽

W.S. Kiser

Keyword(s):

Epithelial Cell ◽

Epithelial Cells ◽

Tubular Epithelial Cell ◽

Tubular Epithelial Cells ◽

Cell Nuclei ◽

Crystalline Structures ◽

Small Crystals ◽

Membrane Bound

During a project in which regenerative changes were studied in autotransplanted canine kidneys, intranuclear crystals were seen in a small number of tubular epithelial cells. These crystalline structures were seen in the control specimens and also in regenerating specimens; the main differences being in size and number of them. The control specimens showed a few tubular epithelial cell nuclei almost completely occupied by large crystals that were not membrane bound. Subsequent follow-up biopsies of the same kidneys contained similar intranuclear crystals but of a much smaller size. Some of these nuclei contained several small crystals. The small crystals occurred at one week following transplantation and were seen even four weeks following transplantation. As time passed, the small crystals appeared to fuse to form larger crystals.

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Membrane-bound vesicles associated with the microvilli in the midgut of the freshwater microcrustacean, Daphnia magna

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100076135 ◽

1983 ◽

Vol 41 ◽

pp. 480-481

Author(s):

Patricia L. Jansma

Keyword(s):

Daphnia Magna ◽

Intestinal Epithelial Cells ◽

Uranyl Acetate ◽

Intestinal Epithelial ◽

Chlamydomonas Reinhardii ◽

Thin Sections ◽

Saturated Water ◽

Cacodylate Buffer ◽

Membrane Bound ◽

Ethanol Series

The presence of the membrane bound vesicles or blebs on the intestinal epithelial cells has been demonstrated in a variety of vertebrates such as chicks, piglets, hamsters, and humans. The only invertebrates shown to have these microvillar blebs are two species of f1ies. While investigating the digestive processes of the freshwater microcrustacean, Daphnia magna, the presence of these microvillar blebs was noticed.Daphnia magna fed in a suspension of axenically grown green alga, Chlamydomonas reinhardii for one hour were narcotized with CO2 saturated water. The intestinal tracts were excised in 2% glutaraldehyde in 0.2 M cacodyl ate buffer and then placed in fresh 2% glutaraldehyde for one hour. After rinsing in 0.1 M cacodylate buffer, the sample was postfixed in 2% OsO4, dehydrated with a graded ethanol series, infiltrated and embedded with Epon-Araldite. Thin sections were stained with uranyl acetate and Reynolds lead citrate before viewing with the Philips EM 200.

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Analysis of the Anterior Secretory Cells of Onchidohs Muricata (Nudibranchia)

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100099490 ◽

1981 ◽

Vol 39 ◽

pp. 614-615

Author(s):

Roy Skidmore

Keyword(s):

Endoplasmic Reticulum ◽

Secretory Granules ◽

Golgi Body ◽

The Body ◽

Secretory Cells ◽

Secretory Vesicles ◽

High Magnification ◽

Large Numbers ◽

Membrane Bound ◽

Sole Of The Foot

The long-necked secretory cells in Onchidoris muricata are distributed in the anterior sole of the foot. These cells are interspersed among ciliated columnar and conical cells as well as short-necked secretory gland cells. The long-necked cells contribute a significant amount of mucoid materials to the slime on which the nudibranch travels. The body of these cells is found in the subepidermal tissues. A long process extends across the basal lamina and in between cells of the epidermis to the surface of the foot. The secretory granules travel along the process and their contents are expelled by exocytosis at the foot surface.The contents of the cell body include the nucleus, some endoplasmic reticulum, and an extensive Golgi body with large numbers of secretory vesicles (Fig. 1). The secretory vesicles are membrane bound and contain a fibrillar matrix. At high magnification the similarity of the contents in the Golgi saccules and the secretory vesicles becomes apparent (Fig. 2).

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An Electron Microscopic Study of the Acute Effects of Hypothalamic Releasing Factors on the Anterior Pituitary Gland

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100100901 ◽

1968 ◽

Vol 26 ◽

pp. 232-233

Author(s):

P.W. Coates ◽

E.A. Ashby ◽

L. Krulich ◽

A. Dhariwal ◽

S. McCann

Keyword(s):

Anterior Pituitary ◽

Microscopic Investigation ◽

Uranyl Acetate ◽

Electron Microscopic Investigation ◽

Electron Microscopic ◽

Tissue Samples ◽

Thin Sections ◽

Dense Membrane ◽

Membrane Bound ◽

Releasing Factors

The morphologic effects on somatotrophs of crude sheep hypothalamic extract prepared from stalk-median eminence were studied by electron microscopy in conjunction with concurrently run bioassays performed on the same tissue samples taken from young adult male Sherman rats.Groups were divided into uninjected controls and injected experimentals sacrificed at 5', 15', and 30' after injection. Half of each anterior pituitary was prepared for electron microscopic investigation, the other half for bioassay. Fixation using collidine buffered osmium tetroxide was followed by dehydration and embedment in Maraglas. Uranyl acetate and lead citrate were used as stains. Thin sections were examined in a Philips EM 200.Somatotrophs from uninjected controls appeared as described in the literature (Fig. 1). In addition to other components, these cells contained moderate numbers of spherical, electron-dense, membrane-bound granules approximately 350 millicrons in diameter.

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A High Resolution Cytochemical Study of Nuclear ATPase in Human Spermatozoa

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100116590 ◽

1975 ◽

Vol 33 ◽

pp. 594-595

Author(s):

A. Sosa ◽

L. Calzada

Keyword(s):

High Resolution ◽

Atpase Activity ◽

Nuclear Membrane ◽

Human Sperm ◽

Sperm Nucleus ◽

Human Spermatozoa ◽

Cytochemical Study ◽

Membrane Bound ◽

Sperm Nuclei ◽

Interchromatin Space

The dependence of nuclear metabolism on the function of the nuclear membrane is not well understood. Whether or not the function of the nuclear membrane is partial or totally responsible of the repressed template activity of human sperm nucleus has not at present been elucidated. One of the membrane-bound enzymatic activities which is concerned with the mechanisms whereby substances are thought to cross cell membranes is adenosintriphosphatase (ATPase). This prompted its characterization and distribution by high resolution photogrammetry on isolated human sperm nuclei. Isolated human spermatozoa nuclei were obtained as previously described. ATPase activity was demonstrated by the method of Wachstein and Meisel modified by Marchesi and Palade. ATPase activity was identified as dense and irregularly distributed granules confined to the internal leaflet of the nuclear membrane. Within the nucleus the appearance of the reaction product occurs as homogenous and dense precipitates in the interchromatin space.

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