scholarly journals DEOXYRIBONUCLEIC ACID CYTOPHOTOMETRY OF STAINED HUMAN LEUKOCYTES I. DIFFERENCES AMONG CELL TYPES

1969 ◽  
Vol 17 (4) ◽  
pp. 249-257 ◽  
Author(s):  
BRIAN H. MAYALL
Keyword(s):  
Visible Light ◽  
Dna Content ◽  
Close Agreement ◽  
Deoxyribonucleic Acid ◽  
Cell Types ◽  
Neutrophilic Granulocytes ◽  
Feulgen Reaction ◽  
Human Leukocytes ◽  
Intensity Measurements ◽  
Individual Human

The deoxyribonucleic acid (DNA) content of individual human leukocytes was estimated cytophotometrically using visible light and spreads stained either with gallocyanin-chrome alum following ribonuclease digestion or with the Feulgen reaction. When the cells were measured on a scanning cytophotometer, significant differences in stain intensity were found among slides. Significant differences also were found among the leukocyte types. In gallocyanin-chrome alum preparations, monocytes measured 16% higher than small lymphocytes and 13% higher than neutrophilic granulocytes. In Feulgen preparations, monocytes measured 4% higher than small lymphocytes and 6% higher than neutrophils. These differences among cell types were independent of donor and stain intensity. Measurements of cells within types and within slides frequently showed close agreement, but it is only in this very limited context that the data are consistent with the hypothesis of DNA constancy. Measurements made on a two-wavelength cytophotometer showed a divergence of only 2.1% relative to similar measurements made on the scanning cytophotometer, which suggests that the differences observed among cells and types are unlikely to be artifacts of the instruments. Over-all, the data indicate either that there is variability in DNA content or that DNA is not being expressed correctly by the measured stain content.

scholarly journals DISCREPANCIES BETWEEN CYTOPHOTOMETRIC FEULGEN VALUES AND DEOXYRIBONUCLEIC ACID CONTENT

1971 ◽  
Vol 19 (3) ◽  
pp. 169-174 ◽  
Author(s):  
K. NOESKE
Keyword(s):  
Functional State ◽  
Acid Content ◽  
Deoxyribonucleic Acid ◽  
Cell Types ◽  
Nuclear Proteins ◽  
Feulgen Reaction ◽  
Template Activity ◽  
Colored Product ◽  
Different Cell Types ◽  
Low Template

Diploid nuclei of different cell types of the granulocytopoietic and erythropoietic series showed different Feulgen values. The highest values were found in the most immature nuclei, and the lowest values in the most mature cells. Extraction of acid-soluble nuclear proteins brought the different values to the same level. Examples of similar findings in other cell types, as described in the literature, are discussed. The Feulgen reaction and its colored product depend on the functional state of the chromatin in such a way that nuclei with high template activity of deoxyribonucleic acid show high Feulgen values whereas mature, dense nuclei with low template activity have lower Feulgen values, although both kinds of nuclei contain identical amounts of deoxyribonucleic acid.

scholarly journals Deoxyribonucleic acid-dependent ribonucleic acid polymerases from normal and polyoma-transformed BHK-21/C13 cells

1975 ◽  
Vol 145 (3) ◽  
pp. 509-516 ◽  
Author(s):  
R J Cooper ◽  
H M Keir
Keyword(s):  
Ionic Strength ◽  
Rna Polymerase ◽  
Dna Content ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Actinomycin D ◽  
Activity Ratio ◽  
Cell Types ◽  
Dna Templates ◽  
Alpha Amanitin

DNA-dependent RNA polymerase (EC 2.7.7.6) ACTIVITIES FROM NORMAL BHK-21/C13 cells and from BHK-21/C13 cells transformed by polyoma virus (PYY cells) were solubilized and fractionated on columns of DEAE-Sephadex. Various properties of the A and B enzymes from the two types of cell were compared. 1. The yields of polymerase relative to the DNA content of the nuclear preparations are similar for both cell types. 2. The ionic-strength optima of polymerases A and B are 12.5 mM and 100mM with respect to (NH4)2SO4 for both cell types. 3. The Mn2+/Mg2+ activity ratio (measured at the respective optimum for each cation) for polymerase A from BHK-21/C13 cells was 1.48 and for the polymerase A from PYY cells was 0.55. The corresponding ratios for polymerase B were 10.11 for BHK-21/C13 cells and 22.75 for PYY cells. 4. Minor differences in the ability of the A polymerases to transcribe native and denatured DNA templates were observed; such differences were not apparent when the B polymerases were compared. 5. All the polymerases were inhibited completely by actinomycin D and by rifampicin AF/013, but not markedly so by rifampicin. Alpha-amanitin inhibited polymerase B but not polymerase A.

Méthode d'étude des quantités d'ADN métaphasique après écrasement de méristèmes radiculaires

Genome ◽  
1992 ◽  
Vol 35 (4) ◽  
pp. 706-708
Author(s):  
C. Le Coq ◽  
C. Guervin ◽  
M. Esclapez ◽  
J. Moret
Keyword(s):  
Dna Content ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Nuclear Dna Content ◽  
Feulgen Reaction ◽  
Root Cells ◽  
Meristematic Root

A method is described for standardized preparation of meristematic root cells treated with colchicine for cytophotométrie analysis of metaphase DNA. Deoxyribonucleic acid has been stained by the Feulgen reaction prior to crushing of the cells.Key words: cytophotometry, Ornithogalum, nuclear DNA content.

scholarly journals Genome size of the European domestic goose (Anser anser domesticus)

2009 ◽  
Vol 89 (4) ◽  
pp. 449-455 ◽  
Author(s):  
K Andraszek ◽  
E Wójcik ◽  
A Grużewska ◽  
E Smalec
Keyword(s):  
Genome Size ◽  
Dna Content ◽  
Cell Types ◽  
Heart Cells ◽  
Feulgen Reaction ◽  
Cell Nuclei ◽  
Anser Anser ◽  
Ovary Cells ◽  
Nucleus Surface ◽  
Chicken Erythrocytes

This work is aimed at determining the C-DNA contained within the nuclei of different types of cells in the domestic goose Anser anser. Cells from the lungs, skin, pancreas, kidney, spleen, liver, heart, brain, blood, ovary and testicle were analysed. Cells from the blood, ovary and testicle were smeared onto microscopic glasses, whereas slides from the other organs and tissues were prepared using the paraffin technique. DNA content, as visualized by the Feulgen reaction using computerized image analysis, was examined in 200 nuclei of every type of cell. Chicken erythrocytes were used as reference material. Different concentrations of chromatin within cell nuclei were observed, from small, dispersed clods to an entirely filled nucleus surface. It was stated that the average C-DNA content in the domestic European goose amounts to 1.306 ± 0.327 pg, which gives goose DNA a length in base pair of 1.277 × 109 ± 0.320 × 109 bp after adjustment. The correlation between nucleus size and the C-DNA content was positive and high. In all cell types it exceeded 0.6. The highest was observed in lung and ovary cells, the lowest in skin and the pancreas. The majority of all cells (57.34%) contain DNA at the range between 1.0 to 1.5 pg, especially those from erythrocytes and the pancreas (82 and 76% respectively). Liver cells demonstrate a tendency toward an amount that is higher than 1.5% of the DNA (78.61% cells). Heart cells reveal a tendency downward (98.99% below 1.5 pg). Less than 1.0 pg of DNA was observed in 17.13% of all examined cells. Key words: Domestic goose, cell, cell nuclei, Feulgen reaction, genome size, DNA mass

Nuclear Morphology and the Ultra-Structural Localization of Deoxyribonucleic Acid Synthesis during Interphase

1969 ◽  
Vol 4 (3) ◽  
pp. 569-582
Author(s):  
GILLIAN R. MILNER
Keyword(s):  
Epithelial Cells ◽  
Dna Synthesis ◽  
Deoxyribonucleic Acid ◽  
Ultrastructural Localization ◽  
Cell Types ◽  
Acid Synthesis ◽  
Lung Fibroblasts ◽  
Nuclear Morphology ◽  
Structural Localization ◽  
Electron Microscope Autoradiography

The ultrastructural localization of deoxyribonucleic acid (DNA) synthesis was studied by electron-microscope autoradiography in human transforming lymphocytes, embryonic lung fibroblasts, epithelial cells and normoblasts. Euchromatin was found to be active in DNA synthesis in all cell types studied, whereas heterochromatin was inactive. However, DNA synthesis was also prominent in the regions where heterochromatin was thought to be decondensing to form euchromatin. Analysis of sequential changes in nuclear morphology of the transforming lymphocyte suggested that there is decondensation of heterochromatin during the S-phase until none is left. In nuclei with no heterochromatin a prominent localization of DNA synthesis was at the nuclear membrane. This sequence of complete decondensation of heterochromatin also seemed likely for fibroblasts and epithelial cells. Normoblasts however showed no stage where the nucleus was wholly euchromatic and it is suggested that in this cell decondensation of heterochromatin for replication is localized and transient.

scholarly journals INTRAMITOCHONDRIAL FIBERS WITH DNA CHARACTERISTICS

1963 ◽  
Vol 19 (3) ◽  
pp. 613-629 ◽  
Author(s):  
Sylvan Nass ◽  
Margit M. K. Nass
Keyword(s):  
Osmium Tetroxide ◽  
Deoxyribonucleic Acid ◽  
Proteolytic Enzymes ◽  
Blue Staining ◽  
Zinc Ions ◽  
Feulgen Reaction ◽  
Toluidine Blue Staining ◽  
Complete Digestion ◽  
Buffer Control ◽  
Dna Characteristics

The effects of proteolytic enzymes, ribonuclease, and deoxyribonuclease upon a fibrous component of chick embryo mitochondria, which was previously shown to have many fixation and staining properties characteristic of the bacterial nucleoplasm, are reported. Pepsin digestion of formaldehyde-fixed tissues removed the membranes and matrices of mitochondria, but a pepsin-resistant fibrous material remained which was heavily stained by uranyl and lead ions. Experiments on a DNA "model system" showed that DNA treated with osmium tetroxide can be depolymerized by deoxyribonuclease. Zinc ions strongly inhibited the depolymerization of DNA. Digestion of osmium tetroxide-fixed tissues (fixed only briefly) with deoxyribonuclease for 1 hour greatly reduced the Feulgen staining of the nuclei, and after 4 hours the Feulgen reaction was completely abolished. The reduction and the disappearance of the Feulgen reaction in nuclei was paralleled by partial to complete digestion of the mitochondrial fibers in the regions studied (after 1 and 4 hours, respectively), without any other obvious changes in cellular structures. When deoxyribonuclease was inhibited by the addition of zinc ions, the nuclear Feulgen reaction was not diminished, nor were the mitochondrial fibers removed. Buffer control incubations for deoxyribonuclease and ribonuclease did not alter the structure or staining properties of the mitochondrial fibers, nor did incubation with ribonuclease. The latter reaction digested the cytoplasmic and nucleolar ribosomes after a 4-hour incubation period, in parallel with the abolishment of toluidine blue staining. The results contribute further evidence that these mitochondria contain deoxyribonucleic acid.

Growth hormone and mammary gland lobule-alveolar development

1961 ◽  
Vol 201 (2) ◽  
pp. 259-263 ◽  
Author(s):  
Richard C. Moon
Keyword(s):  
Growth Hormone ◽  
Mammary Gland ◽  
Dna Content ◽  
Deoxyribonucleic Acid ◽  
Cellular Proliferation ◽  
Estradiol Benzoate ◽  
The Mean ◽  
Alveolar Formation ◽  
Inguinal Glands ◽  
Range Of Values

The effect of growth hormone on mammary gland lobule-alveolar growth in the ovariectomized rat was studied using deoxyribonucleic acid (DNA) of the abdominal-inguinal glands as an index of the degree of cellular proliferation. The administration of 1 mg growth hormone in combination with 2 µg estradiol benzoate for 19 days resulted in alveolar formation and an increase in mammary DNA content above that resulting from injections of either hormone alone. The mean DNA concentration of glands of rats treated with 2 µg estradiol, 6 mg progesterone, 3 µg/100 g l-thyroxine, and 0.5, 1.0, 1.5, and 2.0 mg growth hormone was significantly greater than that of animals receiving only the estradiol, progesterone, and thyroxine. The increase in the mean DNA content was due to a shift in the range of values to a higher plane and did not result from an elevated DNA in only a few animals. It is suggested that the administration of growth hormone during the growth phase of the mammary gland may have a beneficial effect on the subsequent lactation.

Deposition of Lignin in Differentiating Xylem Cell Walls of Normal and Compression Wood of Buxus microphylla var. insularis Nakai

Holzforschung ◽  
1999 ◽  
Vol 53 (2) ◽  
pp. 156-160 ◽  
Author(s):  
Nobuo Yoshizawa ◽  
Hiromi Ohba ◽  
Junko Uchiyama ◽  
Shinso Yokota
Keyword(s):  
Visible Light ◽  
Cell Walls ◽  
Compression Wood ◽  
Outer Region ◽  
Cell Types ◽  
Deposition Process ◽  
The Other ◽  
Normal Wood ◽  
Long Period ◽  
Buxus Microphylla

Summary The deposition process of lignins within differentiating xylem walls of normal and compression wood of Buxus microphylla var. insularis Nakai was examined by visible-light microspectrophotometry coupled with the Wiesner and Mäule reactions. Buxus formed compression wood on the underside of the leaning stems. The secondary walls of the vessels and fibre tracheids in compression wood showed an intense lignification in the outer region of S2 layer. The spectra of tissues after Mäule and Wiesner reactions showed absorption maxima of around 515 nm and 570 nm, respectively. In differentiating xylem cells of normal wood, lignin composed of both guaiacyl and syringyl units was deposited mainly during the S2 thickening and after formation of the S3 layer in fibre tracheids, whereas in vessels it was actively deposited mainly during the S2 thickening. In compression wood, the deposition of the lignin composed of guaiacyl units was observed for a long period from the early stages of the S2 thickening. Lignification was becoming particularly active at the outer portion of S2 layer after completion of the S2 thickening in both vessels and fibre tracheids. On the other hand, the syringyl units were deposited mainly during the S2 thickening in both cell types.

scholarly journals DNA analysis and sorting of viable mouse testis cells.

1981 ◽  
Vol 29 (6) ◽  
pp. 738-746 ◽  
Author(s):  
W M Grogan ◽  
W F Farnham ◽  
J M Sabau
Keyword(s):  
Dna Content ◽  
Adult Mouse ◽  
Dna Analysis ◽  
Cell Types ◽  
Mouse Testis ◽  
Spermatogenic Cells ◽  
Hoechst 33342 ◽  
Cell Sorter ◽  
Pachytene Spermatocytes

The dye Hoechst 33342 and a 2-parameter cell sorter have been used to measure DNA content in viable testis cells and to sort pachytene spermatocytes and round spermatids from adult mouse testis to virtually 100% homogeneity. Early diploid spermatogenic cells were enriched to 90%, a 10-fold purification. The capability for viable sorting of most testis cell types to homogeneity in numbers suitable for many biochemical applications is demonstrated.

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