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.

The development of the tectum in Xenopus laevis: an autoradiographic study

Development ◽  
1972 ◽  
Vol 28 (1) ◽  
pp. 87-115
Author(s):  
K. Straznicky ◽  
R. M. Gaze
Keyword(s):  
Xenopus Laevis ◽  
Dna Synthesis ◽  
Optic Tectum ◽  
Tritiated Thymidine ◽  
Dorsal Surface ◽  
Autoradiographic Study ◽  
Cell Type ◽  
Pole Cells ◽  
Serial Addition ◽  
The Times

The development of the optic tectum in Xenopus laevis has been studied by the use of autoradiography with tritiated thymidine. The first part of the adult tectum to form is the rostroventral pole; cells in this position undergo their final DNA synthesis between stages 35 and 45 or shortly thereafter. Next, the cells comprising the ventrolateral border of the tectum form. These cells undergo their final DNA synthesis at or shortly after stage 45. Finally the cells comprising the dorsal surface of the adult tectum form, mainly between stages 50–55. This part of the tectum originates from the serial addition of strips of cells medially, which displace the pre-existing tissue laterally and rostrally. The formation of the tectum is virtually complete by stage 58. The tectum in Xenopus thus forms in topographical order from rostroventral to caudo-medial. The distribution of labelled cells, several stages after the time of injection of isotope, indicates that, at any one time, a segment of tectum is forming which runs normal to the tectal surface and includes all layers from the ventricular layer out to the surface. In Xenopus, therefore, the times of origin of tectal cells appear to be related not to cell type or tectal layer but to the topographical position of the cells across the surface of the tectum.

scholarly journals Congenital Dyserythropoietic Anemia Type II: Ultrastructural and Radioautographic Studies of Blood and Bone Marrow

Blood ◽  
1972 ◽  
Vol 39 (1) ◽  
pp. 23-30 ◽  
Author(s):  
Kwan Yuen Wong ◽  
George Hug ◽  
Beatrice C. Lampkin
Keyword(s):  
Bone Marrow ◽  
Dna Synthesis ◽  
Marked Decrease ◽  
Tritiated Thymidine ◽  
Red Cells ◽  
Type Ii ◽  
White Girl ◽  
Congenital Dyserythropoietic Anemia ◽  
Cytoplasmic Membranes

Abstract A 12-yr-old white girl with congenital dyserythropoietic anemia (CDA) type II was studied. Excessive cytoplasmic membranes (appearing like "double membranes") were found in the majority of the normoblasts. There was marked decrease in the uptake of tritiated thymidine in the binucleated normoblasts as demonstrated by radioautography. The results suggest that the cells with more severe structural cytoplasmic abnormalities and/or decreased DNA synthesis are destroyed within the bone marrow, and the circulating red cells are derived from a less abnormal population of precursors.

scholarly journals OBSERVATIONS ON THE UPTAKE OF TRITIATED THYMIDINE IN THE PRONUCLEI OF FERTILIZED SAND DOLLAR EMBRYOS

1961 ◽  
Vol 10 (1) ◽  
pp. 59-65 ◽  
Author(s):  
Eva B. Simmel ◽  
David A. Karnofsky
Keyword(s):  
Dna Synthesis ◽  
Nuclear Localization ◽  
Tritiated Thymidine ◽  
Nuclear Material ◽  
Thymidine Uptake ◽  
Sand Dollar ◽  
Male And Female ◽  
Daughter Cells ◽  
Minute Period ◽  
Independent Replication

Following fertilization of the egg of the sand dollar Echinarachnius parma, tritiated thymidine (H3TDR) was taken up independently by the male and female pronuclei beginning within about 15 to 20 minutes, and the labeled pronuclei fused at about 30 to 40 minutes. At cleavage 90 minutes later the labeled nuclear material was distributed to both daughter cells. Unfertilized eggs and sperm exposed to H3TDR did not show nuclear localization of thymidine. DNA replication, thus, is initiated in the haploid pronuclei shortly after fertilization and prior to fusion. The major portion of DNA synthesis, as evidenced by thymidine uptake, appears to be during a 20 to 30 minute period after fertilization. Fertilization is associated with the activation of a mechanism which initiates early and independent replication of DNA in both the male and female pronuclei.

scholarly journals MACROMOLECULAR SYNTHESES RELATED TO THE REPRODUCTIVE CYST OF TETRAHYMENA PATULA

1973 ◽  
Vol 59 (3) ◽  
pp. 615-623 ◽  
Author(s):  
P. R. Gabe ◽  
L. E. de Bault
Keyword(s):  
Protein Synthesis ◽  
Dna Synthesis ◽  
Generation Time ◽  
Rna Synthesis ◽  
Culture Medium ◽  
Tritiated Thymidine ◽  
Growth Cycle ◽  
The Third ◽  
Rna And Protein Synthesis ◽  
The Mean

Macromolecular syntheses in encysted Tetrahymena patula were studied using Feulgen fluorescence cytophotometry, autoradiography, and inhibitors of RNA and protein synthesis. Cycloheximide significantly depressed protein synthesis and D-actinomycin effectively blocked RNA synthesis. Under these conditions, the cells within the cyst were unable to divide. Both cytophotometric measurements and autoradiographic data with tritiated thymidine show that DNA synthesis does not occur during the encystment divisions. Excysted cells placed in nutrient broth medium showed a prolonged generation time after the first cell growth cycle, and by the third generation the mean DNA content per cell was almost triple that of starved excysted cells. These findings indicate that (a) the encystment divisions require RNA and protein synthesis, which are apparently effected through turnover, (b) the encystment division cycles occur in the absence of DNA synthesis, and (c) excysted cells placed in culture medium may go through more than one DNA replication per cell cycle.

scholarly journals TIMING OF DNA SYNTHESIS IN THE MITOTIC CYCLE IN VITRO

1961 ◽  
Vol 9 (3) ◽  
pp. 509-518 ◽  
Author(s):  
Jesse E. Sisken ◽  
Riojun Kinosita
Keyword(s):  
Dna Synthesis ◽  
Generation Time ◽  
Mitotic Cycle ◽  
Tritiated Thymidine ◽  
Lung Cells ◽  
Human Amnion ◽  
Cells In Culture ◽  
Single Colony ◽  
Distribution Of Values

A study was made of the timing of DNA synthesis in the mitotic cycle under conditions where the average mitotic cycle of populations of human amnion and kitten lung cells in culture was variable. Three types of experiments were performed: (a) Autoradiographs were made of incorporated tritiated thymidine in cells whose mitotic histories were recorded microcinematographically allowing the measurement of telophase + G1 along with the total length of the mitotic cycle. (b) Measurement of the G2 + prophase part of the mitotic cycle was performed under various conditions by exposing cells to tritiated thymidine and observing the increase in labeled metaphases plus anaphases as a function of time. (c) The effect of a change in pH on parts of the mitotic cycle was tested by continuously photographing a single colony of cells first at pH 7.8 and then at pH 7.1. All of our data point to the same conclusion; namely, that within a population of cells with a given generation time, the length of each of the measurable parts of the mitotic cycle has a particular distribution of values and that, when there is a change in the generation time, under our conditions only the T + G1 distribution changes.

scholarly journals The generation time, lag time, and minimum temperature of growth of coliform organisms on meat, and the implications for codes of practice in abattoirs

1985 ◽  
Vol 94 (3) ◽  
pp. 289-300 ◽  
Author(s):  
M. G. Smith
Keyword(s):  
Minimum Temperature ◽  
Generation Time ◽  
Time Lag ◽  
Safety Margin ◽  
Lag Time ◽  
Time Limits ◽  
Codes Of Practice ◽  
Generation Times ◽  
Lag Times ◽  
The Times

SUMMARYThe growth of coliform organisms on meat tissue from sheep carcasses processed in a commercial abattoir was investigated. The results indicated that for practical purposes the minimum temperature of growth of these organisms on meat may be taken as 8 °C. Equations were derived relating the generation time and the lag time of coliform organisms in raw blended mutton to the temperature at which the meat is held. Estimates of growth obtained with these equations were found to agree closely with the experimental results, especially at temperatures above 10 °C, and allowed the generation times and the lag times for all temperatures up to 40 °C to be calculated. These times were also found to agree closely with the times determined using a strain ofEscherichia coliinoculated into blended mutton tissue. A strain ofSalmonella typhimuriuminoculated in the same way into blended mutton tissue gave longer generation and lag times at temperatures below 15 °C. Therefore, it is believed that the calculated tables of lag and generation times included in this paper can be used to determine the length of time raw chilled meat may be held afterwards at temperatures above the minimum temperature of growth without an increase in the number of any salmonella organisms present, and these times include a safety margin at each temperature.The study indicates that the mandatory codes of practice presently applied in commercial abattoirs are too stringent. Maintaining the temperature of boning rooms at 10 °C or less does not appear to be necessary providing the meat is processed within the calculated time limits. A relaxation of the restrictions on boning room temperatures would decrease costs, increase worker comfort and safety and would not compromise the bacteriological safety of the meat produced.

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 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.

scholarly journals INTRAPOPULATION KINETICS OF THE MITOTIC CYCLE

1965 ◽  
Vol 25 (2) ◽  
pp. 179-189 ◽  
Author(s):  
Jesse E. Sisken ◽  
Luciano Morasca
Keyword(s):  
Generation Time ◽  
Mitotic Cycle ◽  
Tritiated Thymidine ◽  
Age Distribution ◽  
Time Lapse ◽  
Human Amnion ◽  
Generation Times ◽  
The Right ◽  
Kinetics Of ◽  
Autoradiographic Detection

Data obtained with time lapse cinemicrographic techniques showed that the distribution of generation times for exponentially proliferating human amnion cells in culture is skewed to the right and that reciprocals of generation times appear normally distributed. As shown for bacteria, the true age distribution is much broader than theoretical distributions which fail to take into account the dispersion of generation times. By means of the technique utilizing autoradiographic detection of tritiated thymidine in cells whose mitotic histories were recorded by time lapse cinemicrography, it was shown that the G1 distribution is similar to the generation time distribution but is more variable. In our experiments, the G2 + prophase distribution resembled the generation time and G1 distributions. The data suggested two possibilities for S: either it is relatively constant, or it is inversely related to the lengths of G1 and G2 + prophase. Since G1 is more variable than the total cycle, and G2 + prophase more variable than the computed sum of S + G2 + prophase + metaphase, it was concluded that the relationships between parts of the cycle are non-random and that compensating mechanisms apparently help regulate the lengths of successive parts of the mitotic cycle in individual cells.

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