scholarly journals A STUDY OF NUCLEOLAR VACUOLES IN CULTURED TOBACCO CELLS USING RADIOAUTOGRAPHY, ACTINOMYCIN D, AND ELECTRON MICROSCOPY

1969 ◽  
Vol 43 (2) ◽  
pp. 197-206 ◽  
Author(s):  
J. Morris Johnson
Keyword(s):  
Electron Microscopy ◽  
Normal Level ◽  
Outer Layer ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Control Level ◽  
Outer Edge ◽  
Central Portion ◽  
Tobacco Callus ◽  
Dense Layer

Previously it has been found that in tobacco callus cells nucleolar vacuoles repeatedly form and contract. In this study, nucleolar vacuoles were investigated by using radioautography, actinomycin D, and electron microscopy. It was found, from grain counts of nucleoli labeled with uridine-3H, that nucleoli containing vacuoles had more than three times as many grains/µ2 of nucleolar substance as did nucleolei without vacuoles. Treatment of tobacco callus cells with various concentrations of actinomycin D caused the percentage of cells containing nucleolar vacuoles to decrease; with the highest concentration the percentage of these cells dropped from the normal level of about 70% to less than 10%. However, after removal of actinomycin D the cells regained nucleolar vacuoles up to the control level. When radioautography was used with actinomycin D, it was found that the actinomycin D inhibited the uptake of uridine-3H, i.e. inhibited RNA synthesis, in those nucleoli which lost their nucleolar vacuoles. In addition, after removal of the cells from actinomycin D, it was found that as the cells regained nucleolar vacuoles the nucleoli also began to incorporate uridine-3H. Electron micrographs showed the nucleoli to be composed of a compact, finely fibrous central portion surrounded by a layer of dense particles 100–150 A in diameter. Nucleolar vacuoles occurred in the fibrous central portion. Dense particles similar to those in the outer layer of the nucleoli were found scattered throughout the vacuoles and in a dense layer at their outer edge. These data suggest that in cultured tobacco callus cells the formation and contraction of nucleolar vacuoles is closely related to RNA synthesis in the nucleolus.

scholarly journals REPOPULATION OF POSTMITOTIC NUCLEOLI BY PREFORMED RNA

1973 ◽  
Vol 58 (1) ◽  
pp. 54-63 ◽  
Author(s):  
David M. Phillips ◽  
Stephanie Gordon Phillips
Keyword(s):  
Electron Microscopy ◽  
Cell Cycle ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Normal Morphology ◽  
L929 Cells ◽  
Full Cell ◽  
Nucleolar Rna ◽  
Mitotic Cells

The reconstruction of the nucleolus after mitosis was analyzed by electron microscopy in cultured mammalian (L929) cells in which nucleolar RNA synthesis was inhibited for a 3 h period either after or before mitosis. When synchronized mitotic cells were plated into a concentration of actinomycin D sufficient to block nucleolar RNA synthesis preferentially, nucleoli were formed at telophase as usual. 3 h after mitosis, these nucleoli had fibrillar and particulate components and possessed the segregated appearance characteristic of nucleoli of actinomycin D-treated cells. Cells in which actinomycin D was present for the last 3 h preceding mitosis did not form nucleoli by 3 h after mitosis though small fibrillar prenucleolar bodies were detectable at this time. These bodies subsequently grew in size and eventually acquired a particulate component. It took about a full cell cycle before nucleoli of these cells were completely normal in appearance. Thus, nucleolar RNA synthesis after mitosis is not necessary for organization of nucleoli after mitosis. However, inhibition of nucleolar RNA synthesis before mitosis renders the cell incapable of forming nucleoli immediately after mitosis. If cells are permitted to resume RNA synthesis after mitosis, they eventually regain nucleoli of normal morphology.

THE EFFECTS OF LH-RH ON THE CONCENTRATION OF 3′,5′-CYCLIC AMP AND RNA SYNTHESIS IN RAT ANTERIOR PITUITARY

1975 ◽  
Vol 79 (4) ◽  
pp. 658-662 ◽  
Author(s):  
Takeshi Yoshida ◽  
Yutaka Hattori ◽  
Hiroshi Hoshiai ◽  
Mutsuo Hirano ◽  
Katsuyuki Takahashi ◽  
...  
Keyword(s):  
Cyclic Amp ◽  
Normal Level ◽  
Anterior Pituitary ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Maximum Level ◽  
Tissue Level ◽  
Considerable Extent ◽  
Uridine Uptake ◽  
Lh Rh

ABSTRACT In an experiment performed to investigate the effect of LH-RH on the anterior pituitary of rats, we studied the changes in the concentration of 3′,5′-cyclic AMP in the anterior pituitary and the uridine uptake by the anterior pituitary following the intravenous injection of 100 ng LH-RH. The results obtained are summarized as follows: 1) The tissue level of 3′,5′-cyclic AMP in the anterior pituitary reached the highest peak 15 min after the injection of LH-RH. 2) The uridine uptake of the anterior pituitary began to increase 15 min after the LH-RH injection and attained a maximum level (5 to 6 times the normal level) in 30 min. 3) The fact that the uridine uptake was inhibited to a considerable extent by the addition of actinomycin D to the incubation medium suggested that uridine was incorporated into the RNA fraction.

ULTRASTRUCTURAL EVIDENCE FOR THE STIMULATION OF NUCLEAR RIBONUCLEIC ACID SYNTHESIS BY OESTRADIOL-17β IN THE VAGINAL EPITHELIUM OF THE OVARIECTOMIZED MOUSE

1970 ◽  
Vol 47 (2) ◽  
pp. 143-NP ◽  
Author(s):  
IRINA POLLARD
Keyword(s):  
Ribonucleic Acid ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Control Level ◽  
Staining Technique ◽  
Acid Synthesis ◽  
Vaginal Epithelium ◽  
Ovariectomized Mouse ◽  
Ultrastructural Evidence ◽  
Stimulation Of

SUMMARY The cytochemical nature of the ultrastructural nucleolar transformation previously observed in the vaginal epithelium of the ovariectomized mouse after local application of oestradiol-17β was investigated using a recently developed ultrastructural staining technique. Oestradiol treatment induced ribonucleic acid (RNA) synthesis, especially in the nucleolus but also in the nuclear chromatic region and ribosomes. Actinomycin D administered simultaneously with oestradiol reduced the oestrogen-induced nucleolar response to the control level. These findings are discussed with special reference to the mode of action of oestrogens.

Effect of Some Metabolic Inhibitors on Vinblastine-Induced Ribosomal-Granular Material Complexes

1971 ◽  
Vol 29 ◽  
pp. 548-549
Author(s):  
Awtar Krishan ◽  
Dora Hsu
Keyword(s):  
Granular Material ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Actinomycin D ◽  
Metabolic Inhibitors ◽  
Protein And Rna Synthesis ◽  
Apparent Reduction ◽  
Plant Alkaloids ◽  
In Cells

Cells exposed to antitumor plant alkaloids, vinblastine and vincristine sulfate have large proteinacious crystals and complexes of ribosomes, helical polyribosomes and electron-dense granular material (ribosomal complexes) in their cytoplasm, Binding of H3-colchicine by the in vivo crystals shows that they contain microtubular proteins. Association of ribosomal complexes with the crystals suggests that these structures may be interrelated.In the present study cultured human leukemic lymphoblasts (CCRF-CEM), were incubated with protein and RNA-synthesis inhibitors, p. fluorophenylalanine, puromycin, cycloheximide or actinomycin-D before the addition of crystal-inducing doses of vinblastine to the culture medium. None of these compounds could completely prevent the formation of the ribosomal complexes or the crystals. However, in cells pre-incubated with puromycin, cycloheximide, or actinomycin-D, a reduction in the number and size of the ribosomal complexes was seen. Large helical polyribosomes were absent in the ribosomal complexes of cells treated with puromycin, while in cells exposed to cycloheximide, there was an apparent reduction in the number of ribosomes associated with the ribosomal complexes (Fig. 2).

scholarly journals Mechanism of SARS-CoV-2 polymerase stalling by remdesivir

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Goran Kokic ◽  
Hauke S. Hillen ◽  
Dimitry Tegunov ◽  
Christian Dienemann ◽  
Florian Seitz ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Rna Polymerase ◽  
Binding Site ◽  
Molecular Mechanism ◽  
Active Center ◽  
Rna Synthesis ◽  
Active Form ◽  
Nucleoside Triphosphate ◽  
Rna Dependent Rna Polymerase ◽  
Cryo Electron Microscopy

AbstractRemdesivir is the only FDA-approved drug for the treatment of COVID-19 patients. The active form of remdesivir acts as a nucleoside analog and inhibits the RNA-dependent RNA polymerase (RdRp) of coronaviruses including SARS-CoV-2. Remdesivir is incorporated by the RdRp into the growing RNA product and allows for addition of three more nucleotides before RNA synthesis stalls. Here we use synthetic RNA chemistry, biochemistry and cryo-electron microscopy to establish the molecular mechanism of remdesivir-induced RdRp stalling. We show that addition of the fourth nucleotide following remdesivir incorporation into the RNA product is impaired by a barrier to further RNA translocation. This translocation barrier causes retention of the RNA 3ʹ-nucleotide in the substrate-binding site of the RdRp and interferes with entry of the next nucleoside triphosphate, thereby stalling RdRp. In the structure of the remdesivir-stalled state, the 3ʹ-nucleotide of the RNA product is matched and located with the template base in the active center, and this may impair proofreading by the viral 3ʹ-exonuclease. These mechanistic insights should facilitate the quest for improved antivirals that target coronavirus replication.

scholarly journals Biosynthesis of growth hormone in the rat anterior pituitary gland. Stimulation of biosynthesis in vitro by insulin

1973 ◽  
Vol 134 (4) ◽  
pp. 1103-1113 ◽  
Author(s):  
A. Betteridge ◽  
M. Wallis
Keyword(s):  
Growth Hormone ◽  
Anterior Pituitary ◽  
Rna Synthesis ◽  
Glucose Utilization ◽  
Actinomycin D ◽  
Hormone Synthesis ◽  
Pituitary Glands ◽  
Hormone Biosynthesis ◽  
Stimulation Of

The effect of insulin on the incorporation of radioactive leucine into growth hormone was investigated by using rat anterior pituitary glands incubated in vitro. A 50% stimulation over control values was observed at insulin concentrations above 2μm (280munits/ml). The effect was specific for growth hormone biosynthesis, over the range 1–5μm-insulin (140–700munits/ml). Lower more physiological concentrations had no significant effect in this system. Above 10μm (1.4 units/ml) total protein synthesis was also increased. The stimulation of growth hormone synthesis could be partially blocked by the addition of actinomycin D, suggesting that RNA synthesis was involved. Insulin was found to stimulate the rate of glucose utilization in a similar way to growth hormone synthesis. 2-Deoxyglucose and phloridzin, which both prevented insulin from stimulating glucose utilization, also prevented the effect of insulin on growth hormone synthesis. If glucose was replaced by fructose in the medium, the effect of insulin on growth hormone synthesis was decreased. We conclude that the rate of utilization of glucose may be an important step in mediating the effect of insulin on growth hormone synthesis.

scholarly journals ELECTRON MICROSCOPY OF THE NUCLEAR MEMBRANE OF AMOEBA PROTEUS

1956 ◽  
Vol 2 (4) ◽  
pp. 445-448 ◽  
Author(s):  
Marie H. Greider ◽  
Wencel J. Kostir ◽  
Walter J. Frajola
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Nuclear Membrane ◽  
Outer Layer ◽  
Microscope Study ◽  
Electron Microscope Study ◽  
Cell Types ◽  
Amoeba Proteus ◽  
Nuclear Membranes ◽  
Thin Sectioning

An electron microscope study of the nuclear membrane of Amoeba proteus by thin sectioning techniques has revealed an ultrastructure in the outer layer of the membrane that is homologous to the pores and annuli observed in the nuclear membranes of many other cell types studied by these techniques. An inner honeycombed layer apparently unique to Amoeba proteus is also described.

Hormone-induced differentiation of antheridial branches in Achlya ambisexualis: dependence on ribonucleic acid synthesis

1974 ◽  
Vol 20 (7) ◽  
pp. 977-980 ◽  
Author(s):  
David K. Horowitz ◽  
Peter J. Russell
Keyword(s):  
Sexual Differentiation ◽  
Ribonucleic Acid ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Steroid Hormone ◽  
Acid Synthesis ◽  
Aquatic Fungus ◽  
Induced Differentiation ◽  
Sexual Steroid ◽  
Achlya Ambisexualis

Sexual differentiation in male strains of the aquatic fungus Achlya ambisexualis Raper is induced by antheridiol, a sexual steroid hormone secreted by female strains. Antheridiol-induced initiation of the morphologically distinct antheridial branches in male Achlya is completely prevented when DNA-dependent RNA synthesis is inhibited by actinomycin D. In addition antheridial branch elongation is inhibited to a degree proportional to the concentration of actinomycin D added. Thus, evidence indicates that RNA synthesis is required for antheridiol-induced initiation of antheridial branching and that continued RNA synthesis is required for elongation of antheridial branches.

Review — The Orientation of Cellulose Microfibrils in the cell walls of Tracheids in Conifers

IAWA Journal ◽  
2005 ◽  
Vol 26 (2) ◽  
pp. 161-174 ◽  
Author(s):  
Hisashi Abe ◽  
Ryo Funada
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Outer Layer ◽  
Cell Walls ◽  
Secondary Wall ◽  
Middle Layer ◽  
Cellulose Microfibrils ◽  
Conifer Species ◽  
Predominant Orientation ◽  
Scanning Electron

We examined the orientation of cellulose microfibrils (Mfs) in the cell walls of tracheids in some conifer species by field emission-scanning electron microscopy (FE-SEM) and developed a model on the basis of our observations. Mfs depositing on the primary walls in differentiating tracheids were not well-ordered. The predominant orientation of the Mfs changed from longitudinal to transverse, as the differentiation of tracheids proceeded. The first Mfs to be deposited in the outer layer of the secondary wall (S1 layer) were arranged as an S-helix. Then the orientation of Mfs changed gradually, with rotation in the clockwise direction as viewed from the lumen side of tracheids, from the outermost to the innermost S1 layer. Mfs in the middle layer of the secondary wall (S2 layer) were oriented in a steep Z-helix with a deviation of less than 15° within the layer. The orientation of Mfs in the inner layer of the secondary wall (S3 layer) changed, with rotation in a counterclockwise direction as viewed from the lumen side, from the outermost to the innermost S3 layer. The angle of orientation of Mfs that were deposited on the innermost S3 layer varied among tracheids from 40° in a Z-helix to 20° in an S-helix.

In vitro development of isolated ectoderm from axolotl gastrulae

Development ◽  
1984 ◽  
Vol 80 (1) ◽  
pp. 321-330
Author(s):  
Jonathan M. W. Slack
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Light Microscopy ◽  
Outer Layer ◽  
Microscope Examination ◽  
Animal Pole ◽  
In Vitro Development ◽  
Electron Microscope Examination ◽  
Vitro Development

The development of ectoderm isolated from the animal pole of axolotl gastrulae is monitored by light microscopy, electron microscopy and analysis of newly synthesized proteins, glycoproteins and glycolipids. When control embryos are undergoing neurulation it is shown that the explants autonomously begin to express epidermal markers and do not express mesodermal markers. However the results suggest that not all the cells become epidermal and electron microscope examination shows that only the outer layer does so, the inner cells remaining undifferentiated.

Sign in / Sign up

Export Citation Format

Share Document