Inhibition of Mammalian RNA Synthesis by the Cytoplasmic Ca2+Buffer BAPTA. Analyses of [3H]Uridine Incorporation and Stress-Dependent Transcription†

Biochemistry ◽  
2004 ◽  
Vol 43 (29) ◽  
pp. 9576-9582 ◽  
Author(s):  
Jie Shang ◽  
Mark A. Lehrman
Keyword(s):  
Rna Synthesis ◽  
Uridine Incorporation ◽  
Stress Dependent

Cellular heterogeneity of uridine incorporation in collecting tubules: effect of DOCA

1985 ◽  
Vol 248 (4) ◽  
pp. F552-F564
Author(s):  
A. Vandewalle ◽  
F. Cluzeaud ◽  
M. Chavance ◽  
J. P. Bonvalet
Keyword(s):  
Rna Synthesis ◽  
Cellular Heterogeneity ◽  
Nuclear Surface ◽  
Intercalated Cells ◽  
Uridine Incorporation ◽  
Collecting Tubules ◽  
Silver Grains ◽  
Stimulation Of

In previous studies we showed that in vitro uridine incorporation along the renal tubule is heterogeneous and that DOCA induces a stimulation of RNA synthesis in distal cortical and medullary structures. The present work examines by autoradiography of isolated tubules and renal tissue sections the cellular heterogeneity of the connecting (CNT) and cortical collecting (CCT) tubules after in vivo injection of [3H]uridine in normal and DOCA-treated rabbits. Data confirmed the profile of uridine incorporation along the tubule, which was found in in vitro experiments, and the DOCA-induced stimulation of RNA synthesis. In microdissected CNT and CCT of control kidneys, statistical analysis of the distribution of labeling revealed the presence of two distinct cell populations: one with low labeling (2-3 silver grains per nucleus) and one with high labeling (10-13), which represent 64 and 36%, respectively (CNT), and 74 and 26%, respectively (CCT), of the whole population. Histological data showed that the respective proportions of intercalated cells (29% in CNT; 21% in CCT) and connecting tubule cells (65%) or principal cells (79%) are close to those of the populations with high or low labeling. In addition, autoradiographs on renal sections directly demonstrated that the labeling of intercalated cells (19.3 silver grains/100 micron2 nuclear surface in CNT; 14.7 in CCT) was three times higher than that of connecting (6.6) or principal (5.8) cells. In isolated CNT and CCT, DOCA induced similar absolute increases in the labeling of the two populations. However, the relative increase was more than two times higher in the population with low labeling (+131% in CNT, +210% in CCT) than in the one with high labeling (+71% and +98%). We conclude that cell population of the collecting cortical tubule (CNT and CCT) is heterogeneous with regard to uridine incorporation, reflecting RNA synthesis.

RNA SYNTHESIS IN THE MOUSE OVIDUCT AND UTERUS: VARIATIONS BEFORE, DURING AND AFTER ZYGOTE TRANSPORT

1972 ◽  
Vol 54 (1) ◽  
pp. 15-24 ◽  
Author(s):  
F. H. BRONSON ◽  
T. H. HAMILTON
Keyword(s):  
Reproductive Cycle ◽  
Early Pregnancy ◽  
Rna Synthesis ◽  
Transcriptional Activity ◽  
General Pattern ◽  
Radioactive Precursor ◽  
Uridine Incorporation ◽  
Specific Activities ◽  
Egg Transport

SUMMARY Two experiments were performed to examine variations in whole-organ concentrations of RNA, DNA and protein, and specific activities of RNA in vivo in the mouse oviduct and uterus during the oestrous cycle and early pregnancy. Concentrations of DNA did not vary in the oviduct at any stage of the reproductive cycle in either study. In the first experiment with Yale Swiss mice, the concentration of RNA in the oviduct was highest at pro-oestrus, somewhat lower at mid-day of oestrus, and lowest on days 2, 4 and 12 after insemination. Concentrations of RNA were essentially identical in the oviducts of inseminated and non-inseminated females at mid-day of the day following ovulation. The second experiment, using CF-1 mice, confirmed these results, and, in addition, examined oviducal and uterine uptake and incorporation of [3H]uridine. Incorporation of the radioactive precursor into RNA was highest in the oviduct at mid-day of pro-oestrus, was markedly decreased before ovulation, and remained relatively low during the period of egg transport in both inseminated and non-inseminated mice. Most of the RNA necessary for appropriate oviducal function during zygote transport, therefore, is synthesized well before ovulation, and relatively little is formed during the 3 days of tubal residence. The general pattern of incorporation of [3H]uridine into RNA in the uterus appeared basically similar to that shown by the oviduct, except at mid-pregnancy when transcriptional activity of the oviduct genome was relatively low. The two organs also differed markedly in the magnitude of the pro-oestrous surge in RNA synthesis and accumulation, with the uterus showing a greater increase.

scholarly journals The metabolism of aged seeds. The formation of polyribosomes in germinating field bean ( Vicia faba sp. minor) seeds of different ages

2014 ◽  
Vol 61 (2) ◽  
pp. 203-210
Author(s):  
Kazimierz Zalewski
Keyword(s):  
Dehydrogenase Activity ◽  
Vicia Faba ◽  
Rna Synthesis ◽  
Field Bean ◽  
Embryonic Axes ◽  
Uridine Incorporation ◽  
Different Ages ◽  
Seed Ageing ◽  
Aged Seeds

Total dehydrogenase activity and formation of polyribosomes in embryonic axes and cotyledons of field bean seeds from different harvest years were studied. <sup>3</sup>H-uridine incorporation experiments showed that seed ageing was accompanied by decreased capability for RNA synthesis and polyribosome formation. The embryonic axes of seeds with reduced vigor contained lower levels of polyribosomes.

Some evidence for replication-transcription coupling in Physarum polycephalum

1975 ◽  
Vol 18 (1) ◽  
pp. 27-39
Author(s):  
H. Fouquet ◽  
R. Bohme ◽  
R. Wick ◽  
H.W. Sauer ◽  
K. Scheller
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Dna Sequences ◽  
Physarum Polycephalum ◽  
Rna Synthesis ◽  
Selective Inhibition ◽  
S Phase ◽  
Dna Molecules ◽  
Uridine Incorporation ◽  
G2 Phase

Hydroxyurea, at concentrations of 40–60 mM, selectively and effectively blocked incorporation of thymidine into DNA. Inhibition occurred within 5–10 min of application of the agent when DNA synthesis was in progress, while the onset of replication at the beginning of S-phase and DNA synthesis in G2 phase were not affected. Uridine incorporation into TCA-precipitable material, in the presence of hydroxyurea, was significantly (up to 70%) inhibited in early S-phase of the cell cycle. Selective inhibition of RNA synthesis was confirmed for RNA separated into rRNA-rich and poly(A)-rich RNA fractions and analysed by the 2 kinds of DNA-RNA hybridization reactions. Uridine incorporation into poly (A) RNA was also inhibited under conditions where cycloheximide prevented maturation of nascent DNA molecules in early S-phase. We assume that chromatin which is replicating early DNA sequences may be a more competent template for transcription.

scholarly journals Ribonucleic acids in the embryos of germinating wheat grains of different ripeness

2014 ◽  
Vol 49 (4) ◽  
pp. 423-433 ◽  
Author(s):  
Stanisław Weidner ◽  
Krzysztof Kulka
Keyword(s):  
Rna Synthesis ◽  
Wheat Grain ◽  
Ribonucleic Acids ◽  
Wheat Grains ◽  
Uridine Incorporation ◽  
Rna Content

During 48 h germination of wheat grains of different ripeness the amount of RNA in the germinating embryos doubles, while the rate of synthesis is to a large extent correlated with the level of ribonucleic acids accumulated during the development and ripening of the grain. The highest RNA content was noted in the germs developing from grain harvested at the stage of full ripeness, the lowest - in germs from grain at milk ripeness. Intensity of <sup>3</sup>H-uridine incorporation into the RNA fraction of the germs during 24-h germination also depends on ripeness. Significantly lower RNA synthesis was noted in germinating embryos from wheat grain harvested during milk ripeness as compared to wax or full ripeness.

RNA SYNTHESIS IN THE RAT ANTERIOR HYPOTHALAMUS AND PITUITARY: RELATION TO NEONATAL ANDROGEN AND THE OESTROUS CYCLE

1970 ◽  
Vol 48 (1) ◽  
pp. 125-137 ◽  
Author(s):  
D. F. SALAMAN
Keyword(s):  
Ribosomal Rna ◽  
Anterior Pituitary ◽  
Rna Synthesis ◽  
Specific Activity ◽  
Gradient Centrifugation ◽  
Anterior Hypothalamus ◽  
Oestrous Cycle ◽  
Uridine Incorporation ◽  
Cyclic Changes

SUMMARY RNA from the anterior hypothalamus and anterior pituitary of rats has been labelled by incubation in vitro with [3H]uridine and characterized by density gradient centrifugation. A study of normal females during the oestrous cycle showed cyclic changes in [3H]uridine incorporation into rapidly labelled RNA (rl-RNA) both in the anterior pituitary and hypothalamus. In both tissues the specific activity of RNA was low at dioestrus and high at oestrus and metoestrus. In androgenized females, incorporation into hypothalamic rl-RNA was less than the oestrus—metoestrus level and similar to that at dioestrus, while incorporation into anterior pituitary rl-RNA was similar to the oestrus—metoestrus level and greater than at dioestrus. [3H]Uridine incorporation into ribosomal RNA (r-RNA) of anterior hypothalamus and pituitary was also demonstrated by incubation for 4 h. Under these conditions there was no effect of androgenization on hypothalamic r-RNA, but the specific activity of pituitary r-RNA was greater than normal.

scholarly journals Incorporation of [3H] uridine into the ribonucleic acid of rat uterus during pseudopregnancy and in the presence of I.C.I. 46474 [trans-1-(p-β-dimethylaminoethoxyphenyl)-1,2-diphenylbut-1 -ene]

1970 ◽  
Vol 119 (4) ◽  
pp. 609-613 ◽  
Author(s):  
J. E. O'Grady ◽  
P. J. Heald ◽  
Anne O'Hare
Keyword(s):  
Ribonucleic Acid ◽  
Untreated Control ◽  
Rna Synthesis ◽  
Uterine Tissue ◽  
Rat Uterus ◽  
Pregnant Rat ◽  
Uridine Incorporation ◽  
Important Requirement ◽  
Dose Concentration ◽  
Different Doses

1. The incorporation of [3H]uridine into RNA of rat uterine tissue has been measured during pseudopregnancy and in rats receiving different doses of an anti-implantation compound [trans-1-(p-β-dimethylaminoethoxyphenyl)-1,2-diphenylbut- 1-ene, I.C.I. 46474]. 2. In the uterus of the pseudopregnant rat uridine incorporation into RNA increased markedly on day 3 of pseudopregnancy, remained high until day 5 and decreased sharply by day 6, rising again by day 9. 3. This pattern of change was similar to, but not identical with, that previously found in the pregnant rat. 4. Rats receiving I.C.I. 46474 at a dose concentration below that preventing implantation showed a pattern of RNA synthesis similar to that found in untreated control rats. 5. Rats treated with doses of I.C.I. 46474 sufficient to inhibit implantation revealed a totally different pattern of incorporation of [3H]uridine into uterine RNA. 6. The results are discussed in terms of previous findings with the normally pregnant rat. It is concluded that the increasing uterine RNA synthesis found on day 3 in the pregnant rat is an important requirement for the occurrence of the subsequent implantation.

Vermehrte RNS-Synthese in Makrophagen während der Phagocytose antigenen Materials / Increased RNA Synthesis in Macrophages during Phagocytosis of Antigenic Materials

1969 ◽  
Vol 24 (10) ◽  
pp. 1317-1323 ◽  
Author(s):  
Klaus-Ulrich Hartmann
Keyword(s):  
Nucleic Acids ◽  
Rna Synthesis ◽  
Sucrose Gradient ◽  
Gradient Analysis ◽  
Culture Period ◽  
Cell Populations ◽  
Sheep Erythrocytes ◽  
Uridine Incorporation ◽  
Species Specific

Macrophages have been cultured as uniform cell populations. These cells incorporate uridine but not thymidine into their nucleic acids during the culture period. Sheep erythrocytes in the presence of macrophage cytophilic antibodies were agglutinated to rosettes and phagocytized. Cytophilic antibodies did not have to be species specific; phagocytosis was not complement-dependant. During phagocytosis, incorporation of H3-uridine into the RNA was increased. Sucrose gradient analysis showed that the increase in uridine incorporation was mainly due to increased m-RNA synthesis.

Development of the secondary phloem of the primary root of Pisum

1969 ◽  
Vol 17 (2) ◽  
pp. 199 ◽  
Author(s):  
S Zee ◽  
TC Chambers
Keyword(s):  
Fine Structure ◽  
Rna Synthesis ◽  
Early Stage ◽  
Primary Root ◽  
Sieve Element ◽  
Secondary Phloem ◽  
Uridine Incorporation ◽  
Phloem Parenchyma ◽  
Stage Of Development ◽  
Chromatin Material

The divisional pattern of the cambium in giving rise to the secondary phloem and the associated changes in the fine structure of the cellular components from the initial to the morphologically mature sieve element have been described. The fine structure of the sieve element plastid is of particular interest in that it lacks a well-developed internal membrane system but contains two characteristic inclusion bodies, starch granules (which often break up into smaller units) and protein crystalloids (which invariably show subunits with either one- or two-directional periodicity). Electron microscope autoradiographs were made of cambial cells and their phloem derivatives in tissues exposed either to [3H]thymidine for 24 hr (followed by growth in the absence of tracer for 4 days) or to [3H]uridine for 2-12 hr without a further growth period. The [3H]thymidine labels were specifically lodged in the chromatin material of the cambial initials, the companion cells, the phloem parenchyma cells, and the young sieve elements. During sieve element maturation the [3H]thymidine label became less specifically associated with the chromatin material. The pattern of labelling of the [3H]uridine in addition to being associated with the nucleolus was also associated with the chromatin, the electron-lucent areas of the nucleus, and the cytoplasm of the young sieve element, the companion cell and the phloem parenchyma indicating that these cells were all active in RNA synthesis. In the sieve element the nucleolus disappeared at a very early stage of development. This was associated with a decrease in [3H]uridine incorporation into the nucleus. As the tonoplast of the sieve element disintegrated, [3H]uridine incorporation into both the nucleus and the cytoplasm stopped, which is interpreted as a cessation in RNA synthesis. At no stage in the development of the sieve element was [3H]uridine incorporated into the "slime" material.

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