NUCLEIC ACID SYNTHESIS AND TRANSFER IN NORMAL, SECOND GENERATION CHICK EMBRYO FIBROBLASTS

1961 ◽  
Vol 39 (10) ◽  
pp. 1625-1633 ◽  
Author(s):  
R. Bather ◽  
Elizabeth Purdie-Pepper
Keyword(s):  
Chick Embryo ◽  
Rna Synthesis ◽  
Second Generation ◽  
Acid Synthesis ◽  
Quantitative Estimates ◽  
Cytoplasmic Rna ◽  
Embryo Fibroblasts ◽  
Slight Rise ◽  
Zero Time ◽  
Rna And Dna

The stripping film technique of autoradiography has been used to study some aspects of RNA and DNA metabolism in chick embryo fibroblasts in second generation tissue culture.Approximately one third of the cells incorporated thymidine-H3into DNA in a 20-minute uptake period. The duration of DNA synthesis, the generation time (time elapsing between two successive cell divisions), and the duration of mitosis have been calculated and are very similar to the values obtained for pure strains of hamster cells maintained in culture for several months by another author.RNA synthesis, as measured by uridine-H3incorporation, occurred only in the nucleus and nucleolus. Both sites synthesized RNA simultaneously beginning at zero time. The ratio of the number of grains over the nucleolus to that over the whole nucleus remained constant throughout the uptake of uridine-H3and its transfer to the cytoplasm.A small amount of labelled soluble RNA precursors remain in the nucleus after removal of uridine-H3from the medium. This results in a slight rise in radioactivity of the nucleus after uridine-H3removal. RNA then leaves the nucleus rapidly and appears in the cytoplasm. The half life of RNA in the nucleus is about 4 hours. Some turnover of cytoplasmic RNA seems to occur after 8 hours but quantitative estimates of its rate cannot be made due to changing geometry of the cells as the RNA migrates from the nuclear to the peripheral parts of the cell.Finally, little or no RNA synthesis occurs for a period of about 30 minutes during contraction of the chromosomes in mitosis.

The appearance and quantitation of cytoplasmic ribonucleic acid in the early chick embryo

Development ◽  
1972 ◽  
Vol 28 (2) ◽  
pp. 367-384
Author(s):  
C. C. Wylie
Keyword(s):  
Chick Embryo ◽  
Ribosomal Rna ◽  
Rna Synthesis ◽  
Specific Activity ◽  
Cell Number ◽  
Present Knowledge ◽  
Precursor Pool ◽  
Cytoplasmic Rna ◽  
Labelling Experiment ◽  
Neurula Stage

This paper seeks to extend our knowledge about RNA synthesis in early embryogenesis to the domestic fowl, Gallus domesticus. Using this species for research, apart from increasing our knowledge of higher vertebrate embryology, has certain advantages such as rapid uptake of isotopic precursors and ease of microdissection in culture. The following results are presented: (1) The cell number in the whole chick embryos is shown to be increasing logarithmically between the time of laying and the early neurula stage; with a doubling time of 7·4 h. (2) The onset of ribosomal RNA synthesis has been shown to be during mid-cleavage of the chick embryo, while development is taking place in the oviduct and uterus of the mother. (3) In a cumulative labelling experiment, embryos were labelled at the unincubated-egg stage, allowed to develop to various morphological stages up to neurulation, and their cytoplasmic RNA prepared and analysed by gel electrophoresis. (4) The specific activity of the precursor pool for RNA synthesis was measured at several stages, using the same labelling conditions, and the results were used to quantitate the RNA synthesis from the incorporated radioactivity. (5) Using these techniques, it was found that newly synthesized cytoplasmic RNA accumulates steadily in the whole chick embryo, reaching a level of 104 μg by the early neurula stage. On a per cell basis, however, the amount of newly synthesized cytoplasmic RNA seems to decrease slightly. These findings are discussed in the light of present knowledge about embryos of other vertebrates and certain invertebrates.

Effect of Daunorubicin and Adriamycin on Nucleic ACID Synthesis of Serum Stimulated Mouse Embryo Fibroblasts

1977 ◽  
Vol 63 (1) ◽  
pp. 31-41 ◽  
Author(s):  
Rosanna Supino ◽  
Anna M. Casazza ◽  
Aurelio Di Marco
Keyword(s):  
Dna Synthesis ◽  
Mouse Embryo ◽  
Rna Synthesis ◽  
Calf Serum ◽  
Acid Synthesis ◽  
Anthracycline Antibiotics ◽  
Simple Method ◽  
Mouse Embryo Fibroblasts ◽  
Embryo Fibroblasts

This paper reports the effects of daunorubicin and adriamycin on DNA and RNA synthesis of in vitro cultured mouse embryo fibroblasts (MEF) stimulated by fetal calf serum (FCS). The addition of FCS to quiescent MEF cultures brings about a wave of RNA synthesis, followed by DNA synthesis which starts between 8 and 12 h after change of medium and proceed for up to 24 h. These cells are therefore partially synchronized. The level of DNA synthesis depends on the amount of FCS added. Daunorubicin and adriamycin are almost equally effective in inhibiting DNA synthesis, as well as cell proliferation, which takes place later. Adriamycin is more active than daunorubicin on RNA synthesis. In cultures treated for an 8 h period starting at different times after FCS addition, the highest DNA synthesis inhibition is achieved by treatment during the first 8 h, when DNA synthesis has not yet started. The cellular uptake of daunorubicin is constantly higher than that of adriamycin, in any experimental condition tested. The results show that FCS-stimulated MEF can provide a simple method for studying the effects of anthracycline antibiotics on partially synchronized cells.

Side effects ofRicinus lectin (RCA 120) on nucleic acid synthesis in chick embryo fibroblasts

1983 ◽  
Vol 39 (2) ◽  
pp. 184-185 ◽  
Author(s):  
B. Bernard ◽  
C. Berjonneau ◽  
P. Codogno ◽  
J. Font ◽  
M. Aubery
Keyword(s):  
Nucleic Acid ◽  
Side Effects ◽  
Chick Embryo ◽  
Acid Synthesis ◽  
Nucleic Acid Synthesis ◽  
Embryo Fibroblasts

scholarly journals Lomofungin as an inhibitor of nucleic acid synthesis in Saccharomyces cerevisiae

1974 ◽  
Vol 142 (3) ◽  
pp. 457-463 ◽  
Author(s):  
Michael Cannon ◽  
Antonio Jimenez
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Synthesis ◽  
Rna Synthesis ◽  
Acid Synthesis ◽  
Growth Conditions ◽  
Dna And Rna ◽  
Dna And Rna Synthesis ◽  
Inhibition Of Dna Synthesis ◽  
Rna And Dna Synthesis ◽  
Rna And Dna

1. The antibiotic lomofungin was found to be a potent inhibitor of both DNA and RNA synthesis in Saccharomyces cerevisiae. Under selected growth conditions inhibition of DNA synthesis by the drug preceded inhibition of RNA synthesis. 2. Although in general lomofungin inhibited synthesis of ribosomal RNA and polydisperse RNA more effectively than that of low-molecular-weight RNA, under certain conditions the drug inhibited almost completely synthesis of both 4S and 5S RNA. 3. Inhibition of both RNA and DNA synthesis may be explained if RNA synthesis is required for DNA synthesis in yeast. Alternatively, lomofungin, in addition to interacting with DNA-dependent RNA polymerase, might interfere with a component(s) of the DNA-synthetic apparatus. The drug may thus prove to be of considerable value in studies of DNA synthesis in eukaryotes.

scholarly journals The effect of convulsions induced by flurothyl on ribonucleic acid synthesis in rat cerebral cortex during the recovery phase

1975 ◽  
Vol 152 (3) ◽  
pp. 449-467 ◽  
Author(s):  
C V Wynter ◽  
P Ioannou ◽  
A P Mathias
Keyword(s):  
Cerebral Cortex ◽  
Rna Synthesis ◽  
Sodium Dodecyl ◽  
Acid Synthesis ◽  
Recovery Phase ◽  
Major Effect ◽  
Rat Cerebral Cortex ◽  
Cytoplasmic Rna ◽  
Nuclear Rna ◽  
Double Label

The effect of convulsions, induced by flurothyl, on RNA synthesis in purified unfractionated nuclei and the cytoplasm of rat cerebral cortex was studied by using a double-label technique involving injection of [3H]- and [14C]-orotate intracisternally. 2. Intact RNA was extracted in 80% yield by an enzymic method by using a proteinase in the presence of sodium dodecyl sulphate followed by deoxyribonuclease. Electrophoresis on 1.5% polyacrylamide-0.5% agarose gels revealed the presence of giant nuclear RNA of size up to approx. 300 × 10(6) daltons and mRNA of maximal mol.wt. 9 × 10(6)-16 × 10(6). 3. Nuclear RNA synthesis was decreased to 27% in the first 15 min after convulsions but rapidly increased, so that at 1 1/2 h it was 124% of the control, and at 6 h 147%. 4. Labelling of cytoplasmic RNA was decreased to 15% at 15 min after convulsions but had not recovered to control values by 6 h. 5. Analysis of radioactive gel patterns and the 3H/14C ratio at six time-points (15 min-6h) showed that the major effect was inhibition of the processing of heterogeneous nuclear RNA resulting in a sharp decline in the export of newly synthesized RNA from the nucleus. 6. Cytoplasmic RNA patterns indicated that specific messengers were synthesized at different times during the recovery of the cell after convulsions.

NUCLEIC ACID SYNTHESIS IN X-IRRADIATED THYMOCYTES

1966 ◽  
Vol 44 (6) ◽  
pp. 839-852 ◽  
Author(s):  
D. K. Myers ◽  
Kirsten Skov
Keyword(s):  
Dna Synthesis ◽  
Rna Synthesis ◽  
Acid Synthesis ◽  
X Irradiation ◽  
High Concentrations ◽  
Effects Of Radiation ◽  
Inhibition Of Dna Synthesis ◽  
Thymus Cells ◽  
Rna And Dna

The rate of incorporation of thymidine into the deoxyribonucleic acid (DNA) of rat thymocytes in vitro was not immediately affected by low doses of X-radiation, but became progressively more inhibited as the irradiated cells were incubated at 37 °C for periods of up to 6 hours. Ribonucleic acid (RNA) synthesis deteriorated almost at the same rate as DNA synthesis after X-irradiation in vitro, but protein synthesis was slightly more resistant. Addition of 10–50 mM nicotinamide to the irradiated cell suspensions tended to retard the development of this inhibition, particularly at low temperatures, but high concentrations of nicotinamide were also toxic to these cells. The inhibition of DNA synthesis by X-irradiation appeared to follow a qualitatively similar pattern in thymus, spleen, and regenerating liver in vivo.In addition to the inhibition resulting from degenerative processes in the irradiated cells, DNA synthesis in vitro was directly inhibited by 20–30 kr X-radiation. The effects of radiation on DNA synthesis paralleled to some extent its effects on the gel-forming capacity of the deoxyribonucleoprotein from the thymus cells. It is suggested that the normal synthesis of both RNA and DNA in thymocytes depends on the integrity of the deoxyribonucleoprotein.

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