scholarly journals THE FINE STRUCTURE OF THE DNP COMPONENT OF THE NUCLEUS

1963 ◽  
Vol 16 (1) ◽  
pp. 29-51 ◽  
Author(s):  
Elizabeth D. Hay ◽  
J. P. Revel
Keyword(s):  
Fine Structure ◽  
Dna Synthesis ◽  
Interphase Nucleus ◽  
Deoxyribonucleic Acid ◽  
Tritiated Thymidine ◽  
Proliferating Cells ◽  
Interphase Nuclei ◽  
Ultrathin Sections ◽  
Silver Grains ◽  
Dense Chromatin

In the present investigation, the sites of deoxyribonucleic acid (DNA) synthesis and the fate of labeled deoxyribonucleoprotein (DNP) were studied in autoradiographs of ultrathin sections viewed with the electron microscope. Tritiated thymidine was employed as a label for DNA in the nuclei of proliferating cells of regenerating salamander limbs. In the autoradiographic method reported here, dilute NaOH was used to remove the gelatin of the emulsion after exposure and development. The exposed silver grains are not displaced by this treatment and the resolution of fine structure in the underlying section is greatly improved. Our observations suggest that the DNP component is a meshwork of interconnected filaments 50 to 75 A in diameter, which may be cross-linked to form what Frey-Wyssling would term a "reticular gel." The filamentous DNP meshwork is dispersed throughout the interphase nucleus during DNA synthesis, whereas in chromosomes, which are relatively inert metabolically, the meshwork is denser and is aggregated into compact masses. Dense chromatin centers in interphase nuclei are similar in fine structure to chromosomes and are also inert with respect to DNA synthesis. In the Discussion, the structure of the filamentous meshwork in chromatin is compared with that in chromosomes, and speculations are made as to the functional significance of the variations in DNP fine structure observed.

scholarly journals HETEROCHROMATIN IN HUMAN MALE LEUKOCYTES

1963 ◽  
Vol 16 (2) ◽  
pp. 315-322 ◽  
Author(s):  
A. Lima-de-Faria ◽  
J. Reitalu
Keyword(s):  
Dna Synthesis ◽  
Blood Cells ◽  
Tritiated Thymidine ◽  
White Blood Cells ◽  
Central Position ◽  
Interphase Nuclei ◽  
Human Male ◽  
Chromosome Segments ◽  
Silver Grains ◽  
The Eu

Tritiated thymidine was added to peripheral blood cultures containing phytohemagglutinin so that DNA synthesis in interphase nuclei of white blood cells in the human male could be studied. After 57 hours in culture, a large heterochromatic body with a central position is seen in unlabeled Feulgen-stained nuclei. In labeled nuclei in which DNA synthesis was taking place in both the eu- and heterochromatin at the time the thymidine became available, the heterochromatin shows a higher number of silver grains per unit area, accompanied by a stronger Feulgen reaction, an indication of its higher DNA content. The time of DNA synthesis in the heterochromatin blocks is different from that in the surrounding euchromatin. The large heterochromatic block is composed of chromosome segments gathered together around the nucleolus but it is not part of this organelle. In preparations stained with azure A and acid fuchsin for demonstrating both the nucleolus and the chromosomes, six distinctly heteropyenotic chromosome segments can be seen associated with the nucleolus. Cells of all size categories were found to incorporate tritiated thymidine. The distinct appearance of autosomal heterochromatin in white blood cells may be the result of the new physiological conditions to which the cells are subjected in the medium containing phytohemagglutinin.

scholarly journals DURATION OF PREMEIOTIC DEOXYRIBONUCLEIC ACID SYNTHESIS AND THE STAGES OF PROPHASE I IN RABBIT OOCYTES

1970 ◽  
Vol 47 (3) ◽  
pp. 577-584 ◽  
Author(s):  
J. J. Kennelly ◽  
R. H. Foote ◽  
R. C. Jones
Keyword(s):  
Dna Synthesis ◽  
Meiotic Prophase ◽  
Deoxyribonucleic Acid ◽  
Tritiated Thymidine ◽  
Acid Synthesis ◽  
Prophase I ◽  
Meiotic Prophase I ◽  
Maximum Duration ◽  
Deoxyribonucleic Acid Synthesis ◽  
Silver Grains

To estimate the duration of oocyte DNA synthesis 36, 3-day-old female rabbits received 3, 6, 9, 12, 15, or 18 injections of tritiated thymidine (thy-3H) at hourly intervals. The ovaries, removed at 1, 10, or 20 days after the first injection, were radioautographed. Counts made of the number of silver grains associated with oocyte nuclei in meiotic Prophase I indicate that the duration of DNA synthesis is between 9 and 12 hr. To determine the length of the stages of meiotic Prophase I, a group of 2-3-day-old rabbits was given a single sub-cutaneous injection of thy-3H, and the ovaries were removed at hourly and/or daily intervals after treatment. The minimum duration of leptotene was 3 hr and the maximum duration probably was less than 8 hr. The maximum durations of zygotene, pachytene, and diplotene were estimated to be 44, 216, and 96 hr, respectively. The interval from the end of oogonial DNA synthesis to the beginning ofpremeiotic DNA synthesis (G2 + Mitosis + G1) appeared to be less than 6 hr.

An Ultrastructural and Radio-Autographic Study of the Evolution of the Interphase Nucleus in Plant Meristematic Cells (Allium Porrum)

1974 ◽  
Vol 14 (2) ◽  
pp. 263-287
Author(s):  
J. G. LAFONTAINE ◽  
A. LORD
Keyword(s):  
Interphase Nucleus ◽  
Structural Changes ◽  
Tritiated Thymidine ◽  
Light And Electron Microscopy ◽  
Allium Porrum ◽  
Meristematic Cells ◽  
Fibrillar Material ◽  
Nuclear Structures ◽  
S Period ◽  
Dense Chromatin

Radioautography under both light and electron microscopy was exploited to investigate the structural changes of the chromatin reticulum which characterizes the interphase nucleus of a number of plants. Allium porrum meristematic plant cells were used for this purpose. In this species, the telophase chromosomes uncoil into dense strands which, during the G1 period, gradually give rise to a coarse reticulum. There then follows an extensive unravelling of portions of these strands, and high-resolution radioautography reveals that labelling with tritiated thymidine predominantly occurs over zones of the nucleus consisting of diffuse fine fibrillar material. As the S-period progresses, a chromatin reticulum reappears throughout the nuclear cavity, the tortuous strands being approximately 0.25 µm in diameter. Most of the radioautographic grains still remain over the light nucleoplasmic areas but a number of these are now located on the outermost portion of the dense chromatin profiles. By the end of the S-period, the chromatin strands are slightly thicker (ca. 0.3 µm) and form a looser reticulum. Labelling has decreased noticeably in nuclei of that period, the radioautographic grains being grouped into clusters resting over more or less spherical regions of the chromatin reticulum. Judging from their localization at the surface of the nucleolus or close to the nuclear envelope, these structures correspond to chromocentres. The additional interesting finding that such nuclear structures appear much less compactly organized strongly suggests that chromocentres undergo important conformational modifications during duplication of their DNA.

scholarly journals THE REPLICATION TIME AND PATTERN OF THE LIVER CELL IN THE GROWING RAT

1963 ◽  
Vol 18 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Joseph Post ◽  
Cheng-Ya Huang ◽  
Joseph Hoffman
Keyword(s):  
Dna Synthesis ◽  
Liver Cell ◽  
Tritiated Thymidine ◽  
Liver Cells ◽  
Male Rats ◽  
Interphase Nuclei ◽  
Replication Time ◽  
Wistar Strain ◽  
Growing Rat ◽  
Liver Cell Population

Three-week-old male rats of the Wistar strain were given tritiated thymidine, 1 µc/gm body weight, intraperitoneally and were killed at intervals from 0.25 to 72 hours later. Autoradiographs were made from 5 µ sections, stained by the Feulgen method. The replication time and its component intervals were determined from the scoring of the labeling of interphase nuclei as well as of prophase, metaphase, anaphase, and telophase nuclei. Absorption of the intraperitoneally injected label is rapid and is attended by "flash" labeling during interphase. The results show that at any one time about 4 per cent of the liver cells are synthesizing DNA preliminary to cell division. These cells alternate with waves of other cells and it is estimated that about 10 per cent of the liver cell population is engaged in cell duplication. The replication time is about 21.5 hours, and its component intervals occupy the following times: DNA synthesis, 9 hours; post-DNA synthesis gap, 0.50 hour; prophase, 1.3 hours; metaphase, 1.0 hour; anaphase, 0.4 hour; telophase, 0.3 hour; postmitosis gap, 9.0 hours. A group of liver cells has been recorded in at least 3 successive replication cycles.

scholarly journals SYNTHESIS OF DEOXYRIBONUCLEIC ACID BY MICRO- AND MACRONUCLEI OF TETRAHYMENA PYRIFORMIS

1962 ◽  
Vol 13 (2) ◽  
pp. 193-203 ◽  
Author(s):  
Barbara B. McDonald
Keyword(s):  
Dna Synthesis ◽  
Generation Time ◽  
Deoxyribonucleic Acid ◽  
Tritiated Thymidine ◽  
Tetrahymena Pyriformis ◽  
Type Ii ◽  
Total Period ◽  
Daughter Cells ◽  
Generation Times ◽  
The Times

Evidence as to the times of DNA synthesis in micronucleate Tetrahymena pyriformis (mating type II, variety 1) has been obtained by briefly exposing individuals of different ages to tritiated thymidine, returning them to non-radioactive medium, fixing at division, and preparing autoradiographs. A variable length of interphase, ranging from a few minutes to about 2 hours, has been found to precede the initiation of macronuclear DNA synthesis. Once begun, however, the period of synthesis appears to be similar in all cells, regardless of generation time, and has been estimated at 1 to 1½ hours. Under the conditions of these experiments, the time elapsing between the end of synthesis and subsequent division into daughter cells ranges from approximately 1½ to 2½ hours in generation times long enough to allow such variability. Division of the micronucleus occurs shortly before the cell begins to divide; its DNA synthesis starts immediately and continues after cell division for a total period estimated at about an hour.

scholarly journals Involvement of deoxyribonucleic acid polymerase β in nuclear deoxyribonucleic acid synthesis

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Dna Polymerase Beta ◽  
Cell Nuclei ◽  
High Molecular Weight Dna ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.

A STUDY OF BLASTOCYSTS DURING DELAY AND SUBSEQUENT IMPLANTATION IN LACTATING MICE

1968 ◽  
Vol 42 (3) ◽  
pp. 453-463 ◽  
Author(s):  
ANNE McLAREN
Keyword(s):  
Dna Synthesis ◽  
Thymidine Incorporation ◽  
Tritiated Thymidine ◽  
Uterine Horn ◽  
Uterine Epithelium ◽  
Serial Sections ◽  
Uterine Lumen ◽  
Fresh State

SUMMARY Blastocysts were studied on the 5th and 8th day of pregnancy in lactating mice, in the fresh state, flushed from the uterus, in squash preparations and in serial sections. At the earlier period some mitosis was observed. Tritiated thymidine incorporation studies gave some evidence of DNA synthesis on the 5th and 6th days of pregnancy. By the 8th day the blastocysts were longer, contained more cells, and mitosis had ceased. They were located at the anti-mesometrial end of the uterine lumen, closely apposed to the uterine epithelium, and with their long axes parallel to the long axis of the uterine horn. Implantation could be induced, either by the removal of the litter, or by the injection of an appropriate dose of oestrogen on the 5th or 7th (but not the 4th) day of pregnancy. Both treatments were followed by the appearance of W-bodies in the neighbourhood of the blastocysts, the disappearance of the shed zonae, and the appearance of Pontamine Blue reactivity, oedema of the uterine stroma and formation of the primary decidual zone, in that order.

DEOXYCYTIDYLATE DEAMINASE LEVELS IN REGENERATING RAT LIVER

1961 ◽  
Vol 39 (6) ◽  
pp. 1043-1054 ◽  
Author(s):  
D. K. Myers ◽  
C. Anne Hemphill ◽  
Constance M. Townsend
Keyword(s):  
Dna Synthesis ◽  
Liver Regeneration ◽  
Partial Hepatectomy ◽  
Rat Liver ◽  
Deoxyribonucleic Acid ◽  
Regenerating Liver ◽  
Regenerating Rat Liver ◽  
High Level ◽  
Early Regeneration

Deoxycytidylate deaminase activity and net synthesis of deoxyribonucleic acid (DNA) in vivo were found to increase at approximately the same time during the early stages of liver regeneration. However, deaminase activity in the regenerating liver remained at a high level for 1 day after DNA synthesis had slowed down again during the later stages of regeneration. The increase in deaminase activity was restricted as a result of exposure to 600 r X radiation during early regeneration, but this effect only became evident 11–16 hours after the irradiation. Irradiation on the second day after partial hepatectomy, when deaminase levels in control regenerating livers were relatively constant, failed to affect the deaminase activity immediately but did produce a 40–50% decrease in activity 11–16 hours later. Other antimitotic agents, e.g., colchicine, had little effect on deaminase activity.

scholarly journals Primase p49 mRNA expression is serum stimulated but does not vary with the cell cycle.

1989 ◽  
Vol 9 (5) ◽  
pp. 1940-1945 ◽  
Author(s):  
B Y Tseng ◽  
C E Prussak ◽  
M T Almazan
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Mammalian Cells ◽  
Cell Cycle Progression ◽  
Small Subunit ◽  
Mrna Levels ◽  
Proliferating Cells ◽  
Restriction Point ◽  
Serum Stimulation ◽  
G1 Cells

Expression of the small-subunit p49 mRNA of primase, the enzyme that synthesizes oligoribonucleotides for initiation of DNA replication, was examined in mouse cells stimulated to proliferate by serum and in growing cells. The level of p49 mRNA increased approximately 10-fold after serum stimulation and preceded synthesis of DNA and histone H3 mRNA by several hours. Expression of p49 mRNA was not sensitive to inhibition by low concentrations of cycloheximide, which suggested that the increase in mRNA occurred before the restriction point control for cell cycle progression described for mammalian cells and was not under its control. p49 mRNA levels were not coupled to DNA synthesis, as observed for the replication-dependent histone genes, since hydroxyurea or aphidicolin had no effect on p49 mRNA levels when added before or during S phase. These inhibitors did have an effect, however, on the stability of p49 mRNA and increased the half-life from 3.5 h to about 20 h, which suggested an interdependence of p49 mRNA degradation and DNA synthesis. When growing cells were examined after separation by centrifugal elutriation, little difference was detected for p49 mRNA levels in different phases of the cell cycle. This was also observed when elutriated G1 cells were allowed to continue growth and then were blocked in M phase with colcemid. Only a small decrease in p49 mRNA occurred, whereas H3 mRNA rapidly decreased, when cells entered G2/M. These results indicate that the level of primase p49 mRNA is not cell cycle regulated but is present constitutively in proliferating cells.

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