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.

scholarly journals INHIBITING EFFECT OF THE NEW CYTOTOXIC ANTIBIOTIC DAUNOMYCIN ON NUCLEIC ACIDS AND MITOTIC ACTIVITY OF HELA CELLS

1965 ◽  
Vol 27 (3) ◽  
pp. 545-550 ◽  
Author(s):  
A. Di Marco ◽  
R. Silvestrini ◽  
S. Di Marco ◽  
T. Dasdia
Keyword(s):  
Mitotic Activity ◽  
Rna Synthesis ◽  
Thymidine Incorporation ◽  
Acid Synthesis ◽  
Acetyl Derivative ◽  
Total Inhibition ◽  
Nuclear Rna ◽  
Nucleolar Rna ◽  
Higher Doses

The effect has been studied of Actinomycin D, Daunomycin (Da.), and Da. N acetyl derivative on mitotic activity and on the nucleic acid synthesis of in vitro HeLa cell cultures. The experiments were carried out by means of the radioautographic technique using stripping films. The relative uptake of thymidine-H3 and uridine-H3 was determined by means of the reduced silver grain count present in the nuclei of controls and treated cells. The mitotic activity and thymidine incorporation were noticeably reduced by Daunomycin and Actinomycin, whereas both processes appeared less affected by Da. N acetyl derivative. As regards nuclear RNA synthesis, all three antibiotics at low doses chiefly inhibit nucleolar RNA synthesis. On the other hand, whilst Actinomycin at higher doses causes an almost total inhibition of the synthesis of the whole nuclear RNA, in Daunomycin- and Da. N acetyl derivative-treated cells extranucleolar RNA synthesis is less susceptible to inhibition.

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.

Uptake of [3H]Uridine into Precursor Pools and RNA in Osteogenic Cells

1967 ◽  
Vol 2 (1) ◽  
pp. 39-56
Author(s):  
MAUREEN OWEN
Keyword(s):  
Protein Synthesis ◽  
Mammalian Cells ◽  
Rna Synthesis ◽  
Degradation Products ◽  
Cell Types ◽  
Water Soluble ◽  
Radioactive Label ◽  
Cytoplasmic Rna ◽  
Nuclear Rna

Young rabbits were given a single intraperitoneal injection of [3H]uridine. Using the technique of water-soluble autoradiography a study was made of the uptake of the radioactive label into soluble precursors and RNA in cells on an actively growing bone surface. Labelling of the soluble intracellular pools was immediate, but incorporation of label from these pools into RNA was not completed until 24 h after injection. At this time all the label in the sections was in RNA but this represented only 30% of the total label initially in the soluble pools. This means that 70% of the label is lost from the cell in the first 24 h either as degradation products of RNA synthesis or by other as yet unknown mechanisms. The pattern of labelling of the RNA was similar to that previously found for other mammalian cells in vivo or in vitro. There was a rapid uptake of label into nuclear RNA which reached a maximum by 2 h after injection and a slower uptake into cytoplasmic RNA which reached a maximum by 24 h after injection. There was a slow loss of label from the cells after 24 h indicating a half-life of about 8 days for this relatively stable RNA. A comparison was made of RNA synthesis in the proliferating preosteoblasts and the highly differentiated non-dividing osteoblasts. Labelling of the nuclear RNA for the two cell types was identical. The rate of labelling of the cytoplasmic RNA was similar for the two cell types but the maximum level of labelling in the cytoplasm of the osteoblasts was 2 to 3 times that in the preosteoblasts. This could be correlated with the more active protein synthesis by the osteoblasts. There was a slow loss of labelled RNA by the osteoblasts and preosteoblasts and a rapid loss by the osteocytes after the cells had been incorporated within the bone. It was suggested that this loss paralleled the decline in the rate of protein synthesis by the cells as their environment changed.

scholarly journals The diglyceride kinase of rat cerebral cortex

1971 ◽  
Vol 122 (2) ◽  
pp. 171-179 ◽  
Author(s):  
E. G. Lapetina ◽  
J. N. Hawthorne
Keyword(s):  
Cerebral Cortex ◽  
Phosphatidic Acid ◽  
Nerve Ending ◽  
Osmotic Shock ◽  
Sodium Deoxycholate ◽  
Acid Synthesis ◽  
Optimum Concentration ◽  
Subcellular Fractions ◽  
Rat Cerebral Cortex ◽  
Triton X

1. Formation of phosphatidic acid by diglyceride kinase (EC 2.7.1.-) in the presence of ATP and Mg2+ was shown in a homogenate and subcellular fractions of rat cerebral cortex. 2. The kinase was activated by Mg2+. Ca2+ activated to a smaller extent but was inhibitory in the presence of optimum concentration of Mg2+. Activity was greatly increased in the presence of added 1,2-diglyceride. 3. Sodium deoxycholate markedly stimulated the reaction, but other detergents (Cutscum and Triton X-100) did not. 4. Diglyceride kinase was concentrated in the supernatant and microsomal fractions from rat cerebral cortex. The distribution of the kinase in the particulate fractions resembled that of acetylcholinesterase and 5′-nucleotidase. 5. The rate of phosphatidic acid synthesis by the diglyceride kinase route was much greater than reported rates for acylation of 3-glycerophosphate and was also very rapid in comparison with the rates of other steps in the synthesis of phosphoinositides. 6. Acetylcholine had no stimulatory effect on diglyceride kinase of isolated intact nerve-ending particles or of nerve-ending membranes obtained after osmotic shock.

The measurement of the production of during erythroid differentiation of the Friend erythroleukemia cell

1987 ◽  
Vol 65 (3) ◽  
pp. 188-194 ◽  
Author(s):  
E. Schmedt ◽  
L. Kleiman
Keyword(s):  
Rna Synthesis ◽  
Rna Polymerase Iii ◽  
Erythroid Differentiation ◽  
Gene Probe ◽  
5S Rna ◽  
Cytoplasmic Rna ◽  
Nuclear Rna ◽  
Nuclear Synthesis

The production of [Formula: see text] during Friend cell erythroid differentiation has been studied. In vitro measurements of total nuclear RNA synthesis in nuclei isolated from Friend cells at different stages of differentiation show the total RNA synthesis increases 1.5-fold at day 1 of induction and then decreases through days 2 and 3 to approximately 75% of its rate of synthesis in the nuclei of uninduced cells. The synthesis of RNA polymerase III transcripts undergoes a similar fluctuation through day 2 of induction, but increases again at day 3. The specific synthesis of [Formula: see text] was measured by hybridization of labelled nuclear RNA to a [Formula: see text] gene probe. During erythroid differentiation the percentage of nuclear RNA represented by [Formula: see text] remains constant (0.065%), so that the absolute synthesis of [Formula: see text] fluctuates during differentiation, in parallel with the fluctuations in the synthesis of total nuclear RNA. The relative synthesis of [Formula: see text]in vivo was studied by labelling cells with 35Pi, isolating the resulting radioactive tRNA – 5S RNA population, and hybridizing this population to a [Formula: see text] gene probe. The ratio of [Formula: see text] in newly synthesized cytoplasmic RNA remains similar throughout differentiation (averaging 0.0171), implying that the fluctuations observed in the nuclear synthesis of [Formula: see text] during differentiation probably also occur for the nuclear synthesis of most tRNA and 5S RNA species. Attempts were made to measure the relative steady-state concentration of [Formula: see text] using both aminoacylation and in vitro end labelling of tRNA followed by hybridization to a [Formula: see text] gene probe. These two methods gave different results and we discuss the possible pitfalls of using enzymatic methods for quantitating tRNA concentrations in the cell.

Translational Control of Protein Synthesis in Eat Anterior Pituitary by N-2′-O-dibutyryl adenosine 3′,5′-monophosphate

1973 ◽  
Vol 51 (6) ◽  
pp. 913-919 ◽  
Author(s):  
Roger Boucher ◽  
Marie Gauthier ◽  
Paul Jolicoeur ◽  
Fernand Labrie
Keyword(s):  
Protein Synthesis ◽  
Translational Control ◽  
Rna Synthesis ◽  
Actinomycin D ◽  
Short Term ◽  
Nucleotide Pools ◽  
Cytoplasmic Rna ◽  
Nuclear Rna ◽  
Preferential Incorporation ◽  
Stimulation Of

Actinomycin D, at doses (25 and 50 μg/ml) that block RNA synthesis to less than 3% of the control rate, inhibits the incorporation of [3H]leucine into adenohypophyseal proteins and the release of newly synthesized proteins by 50 and 60%, respectively, of the control rates. Despite this lowering of basal levels of total protein synthesis and release in presence of the antibiotic, the percentage of stimulation of both protein synthesis and release by 5 mM N6-2′-O-dibutyryl adenosine 3′5′-monophosphate (dbcAMP) is not depressed by actinomycin D. When rat hemipituitaries are incubated with [3H]uridine, dbcAMP does not stimulate the labeling of total cytoplasmic RNA or the preferential labeling of any cytoplasmic RNA species resolved on sucrose gradient. There is no stimulatory effect of dbcAMP on total labeling or preferential incorporation into nuclear RNA species extracted at 24 °C or at 65 °C. Labeling of the nucleotide pools was unchanged up to 1 h of incubation but was increased (40–70%) during the last [Formula: see text] of incubation. These data suggest that the short-term stimulatory effects of dbcAMP on total adenohypophyseal protein synthesis and release are exerted at the transiational level.

Über die Beziehungen der RNS- und Proteinsynthese zum Zellkern in Siebzellen von Vicia faba

1964 ◽  
Vol 19 (11) ◽  
pp. 1066-1071 ◽  
Author(s):  
Stefanie Neumann ◽  
Reinhold Wollgiehn
Keyword(s):  
Vicia Faba ◽  
Rna Synthesis ◽  
The Other ◽  
Other Hand ◽  
Cytoplasmic Rna ◽  
Nuclear Rna ◽  
Sieve Tubes

Autoradiographic investigations on the dependency of the RNA and protein syntheses on the nucleus have been carried out in sieve tubes of Vicia faba.In young nucleated sieve tubes 3H-uridine is incorporated primarily into the nuclear RNA and later into the cytoplasmic RNA. In sieve tubes with degenerated nuclei no incorporation of uridine in RNA takes place. On the other hand, 3H-phenylalanine is incorporated also into the proteins of old sieve tubes without RNA synthesis.

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