The effect of double-strength standard saline citrate on silver staining. I. Nucleoli and micronucleoli in the somatic and germ line of the grasshopper Arcyptera fusca (Orthoptera)

1986 ◽  
Vol 28 (2) ◽  
pp. 219-226 ◽  
Author(s):  
J. Gosàlvez ◽  
J. de la Torre ◽  
C. Garcia de la Vega ◽  
C. López-Fernández
Keyword(s):  
Silver Nitrate ◽  
Silver Staining ◽  
Germ Line ◽  
Standard Technique ◽  
Previous Treatment ◽  
Nucleolar Activity ◽  
Standard Saline Citrate ◽  
Nucleolar Proteins ◽  
In Cells

The nucleolar activity in cells from both somatic and germ line of the grasshopper Arcyptera fusca has been analyzed by means of silver staining using a standard technique with and without a previous treatment of the slides with double-strength standard saline citrate (2 × SSC). Results show that the treatment with 2 × SSC improves the silver staining of the main nucleoli and in some tissues reliably uncovers other sites along the chromosomes with attached silver precipitates (micronucleoli). The parallel origin of the main nucleoli and the micronucleoli, the possible enlargement of rDNA activity in some tissues, and the ability of the silver nitrate to stain certain nucleolar proteins are discussed.Key words: Arcyptera, Orthoptera, nucleolus, micronucleus, silver staining.

Cytogenetic study of silver-staining NOR in 8-cell-stage mouse blastomeres fused to 1-cell-stage embryos

Development ◽  
1988 ◽  
Vol 104 (3) ◽  
pp. 453-463
Author(s):  
A.P. Dyban ◽  
K. Lee ◽  
G.T. O'Neill ◽  
S. Speirs ◽  
M.H. Kaufman
Keyword(s):  
Silver Nitrate ◽  
Silver Staining ◽  
Cytogenetic Study ◽  
Metaphase Plate ◽  
Rrna Synthesis ◽  
Cell Nuclei ◽  
Interphase Nuclei ◽  
Cell Stage ◽  
Nucleolar Activity ◽  
Silver Nitrate Staining

Isolated blastomeres from 8- to 16-cell-stage embryos were fused by standard micromanipulatory means with either unfertilized eggs or fertilized or haploid parthenogenetically activated pronuclear-stage embryos. The hybrid eggs/embryos were incubated overnight in the presence of Colcemid until they had entered the first cleavage division. Air-dried chromosome preparations were then stained with silver nitrate in order to detect active nucleolar organizing regions (NOR). While control unfertilized eggs and 1-cell-stage fertilized and parthenogenetically activated embryos showed no evidence of silver-staining NOR-positive regions, the metaphase plates from 8- to 16-cell embryos showed characteristic NOR-positive regions, while their interphase nuclei also showed a characteristic reticular staining appearance. When hybrids between blastomere nuclei and unfertilized eggs were examined, none of the blastomere nuclei entered mitosis. However, when hybrids between blastomere nuclei and fertilized embryos were examined, in two thirds of the embryos, a single blastomere-derived diploid metaphase plate was present in association with two pronuclear-derived haploid metaphase plates. In most instances, the blastomere-derived chromosomes did not display silver-nitrate-staining NOR. Similar findings were observed when the blastomere-derived chromosomes in hybrids between blastomere nuclei and haploid parthenogenetic embryos were analysed. In the majority of cases, when blastomere nuclei remained in interphase, the characteristic silver-nitrate-staining fine reticular material either was not seen, or the nuclear contents were dispersed into clumps of chromatin-like material. Occasionally, the diploid chromosomes in the hybrids displayed morphological abnormalities. Our findings suggest that the cytoplasm of activated (but not nonactivated) 1-cell embryos is capable of influencing the nucleolar activity of the introduced 8- to 16-cell nuclei, effectively erasing from their chromosomes the memory of at least three previous rounds of rRNA synthesis.

scholarly journals Quantification of Ag-NOR proteins using Ag-NOR staining on western blots.

1994 ◽  
Vol 42 (11) ◽  
pp. 1513-1517 ◽  
Author(s):  
P Roussel ◽  
V Sirri ◽  
D Hernandez-Verdun
Keyword(s):  
Mammalian Cells ◽  
Protein A ◽  
Ribosomal Genes ◽  
Western Blots ◽  
Nucleolar Activity ◽  
Cell Extracts ◽  
Nucleolar Proteins ◽  
Pathological Conditions ◽  
Nuclear Extracts ◽  
Protein B23

Ribosomal genes are associated with a set of silver-stained nucleolar proteins, the Ag-NOR proteins, whose amount is directly related to the duration of the cell cycle. Quantification of Ag-NOR proteins by image analysis is presently used to evaluate the rate of proliferation of cancer cells and nucleolar activity. Our objective was to establish a procedure to quantify independently each major Ag-NOR protein in cell extracts. Computerized densitometry established that the specific silver staining of Ag-NOR proteins (Ag-NOR staining) performed on Western blots makes it possible to quantify Ag-NOR proteins. Using purified Ag-NOR proteins, nucleolin, and protein B23, we observed that the intensity of Ag-NOR staining is proportional to the amount of protein. A linear relationship exists between the intensity of Ag-NOR staining and the amount of nucleolin, in the range of 0.2-1.6 micrograms. Using total nuclear extracts prepared from mammalian cells, the proportionality was maintained for total Ag-NOR-stained proteins or for a particular protein. We also determined the levels of nuclear proteins suitable for quantitative analysis. Individual Ag-NOR proteins can be quantified by computerized densitometry in nuclear extracts after Ag-NOR staining on Western blots. This procedure can be applied to establish the contribution of each Ag-NOR protein in general staining, estimate the variability of each Ag-NOR protein in normal and pathological conditions, and quantify each Ag-NOR protein contained per cell.

New approaches to the role of sulfhydryl groups in silver stainability of protein in grasshopper chromosomes

Genome ◽  
1988 ◽  
Vol 30 (2) ◽  
pp. 133-137 ◽  
Author(s):  
J. de la Torre ◽  
J. L. Bella ◽  
C. López-Fernandez ◽  
J. Gosálvez
Keyword(s):  
Silver Staining ◽  
Chemical Nature ◽  
Germ Line ◽  
Staining Technique ◽  
Sulfhydryl Groups ◽  
Sh Groups ◽  
New Approaches ◽  
Silver Staining Technique ◽  
Reduced State

Silver staining in somatic and germ line cells of the grasshopper Arcyptera fusca has been analyzed after a standard silver staining technique was used with pretreatments that included dithiothreitol and β-mercaptoethanol as reagents to maintain —SH groups in the reduced state. The results show that in some tissues, these pretreatments improve not only the silver staining of nucleolar masses but also the recognition of other silver spots associated with the chromatin. These observations are similar to those previously described for the effect of double-strength standard saline citrate on silver staining. The possible chemical nature of the protein groups responsible for the differential silver stainability and the role of saline citrate in the modification of argyrophylic proteins suggested previously are briefly discussed.Key words: Orthoptera, silver staining, sulfhydryl groups.

Silver Staining of Ribosomal Proteins

1971 ◽  
Vol 9 (1) ◽  
pp. 253-269
Author(s):  
J. W. SMITH ◽  
R. J. STUART
Keyword(s):  
Silver Nitrate ◽  
Perchloric Acid ◽  
Silver Staining ◽  
Ribosomal Proteins ◽  
Nuclear Staining ◽  
Rna Molecules ◽  
Mitochondrial Ribosomes ◽  
Associated Proteins ◽  
Relationship Of ◽  
Formalin Fixed

The effects of staining several tissues with silver nitrate were studied in the electron microscope. Tissues were fixed in 2% glutaraldehyde buffered to pH 7.3 and containing 0.22 M sucrose, some being post-osmicated. Staining was effected by immersion of Araldite sections in unbuffered 5% silver nitrate for 30 min. Increase in electron density was restricted to all nucleic acid-containing structures except mitochondrial DNA which is probably not associated with histones. Treatment of fixed tissues with cold 10% perchloric acid for 24 h, which extracts 85% of the total cell RNA, abolished silver staining within the nucleolus but did not affect that of cytoplasmic or mitochondrial ribosomes. Incubation of isolated, formalin-fixed liver cells with DNase I did not abolish nuclear staining. This evidence suggests that silver nitrate stains selectively the proteins associated with nucleic acids: the abolition of nucleoler staining by perchloric acid is not at present understood but may be due to some difference in relationship of protein to RNA in this, compared with other situations. Silver staining indicates that the loci occupied by RNA-associated proteins differ in size and number in different type of ribosomes. Cytoplasmic ribosomes probably contain 5 loci of 4-6 nm diameter, 2 being situated in the 60-S subunit and 3 in the 40-S subunit. Mitochondrial ribosomes appear to contain 2 smaller loci. In the nucleolus the majority of the ribosomes constituting the granulosa contain 2 unequal loci of about 4 and 6 nm respectively, while the appearance of the pars fibrosa is compatible with the view that it consists of a network of long, extended RNA molecules with which small 2.5-nm protein loci are associated.

Localization of ribosomal RNA (rRNA), rRNA genes and silver staining nucleolar proteins in in vitro produced bovine embryos

1998 ◽  
Vol 49 (1) ◽  
pp. 190
Author(s):  
D. Viuff ◽  
B. Avery ◽  
G. Vajta ◽  
T. Greve ◽  
H. Callesen ◽  
...  
Keyword(s):  
Ribosomal Rna ◽  
Silver Staining ◽  
Rrna Genes ◽  
Bovine Embryos ◽  
Nucleolar Proteins

Specific Aspartic Acid-Rich Sequences Are Responsible for Silver Staining of Nucleolar Proteins

1995 ◽  
Vol 207 (2) ◽  
pp. 485-491 ◽  
Author(s):  
B.C. Valdez ◽  
D. Henning ◽  
T.V. Le ◽  
H. Busch
Keyword(s):  
Aspartic Acid ◽  
Silver Staining ◽  
Nucleolar Proteins

Anti-Tumour Effect of Zoledronate in Cells from B Chronic Lymphoytic Leukemia and Low-Grade Lymphoma Patients.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4830-4830
Author(s):  
Deborah Vidal Vasconcellos ◽  
Karina Lani Silva ◽  
Eric Delfraro de Paula Castro ◽  
Arthur Coelho Moellman ◽  
Maria do Socorro Pombo-de-Oliveira ◽  
...  
Keyword(s):  
Annexin V ◽  
Previous Treatment ◽  
Lymphocytic Leukemia ◽  
Low Grade ◽  
Complementary Role ◽  
Apoptotic Effect ◽  
Previously Treated ◽  
Annexin V Assay ◽  
Anti Tumour Effect ◽  
In Cells

Abstract Recent studies demonstrate that bisphosphonates - anti-resorptive drugs - have direct anti-tumour effect in many tumour cell lines, including hematopoietic ones. The aim of this study was to evaluate the apoptotic effect of Zoledronate in cells from B chronic lymphocytic leukemia (B-CLL) and low-grade lymphoma (LGL) patients. Samples of 19 B-CLL or LGL patients were incubated with Zoledronate 100 μM in RPMI medium supplemented with fetal bovine serum 10% at 370C for 12h. Apoptosis using Annexin V assay, by flow cytometry (FC), was observed in 8 out of 19 (42,1%) patients despite previous treatment. Multidrug resistance (MDR) phenotype was performed using rhodamine-123 efflux assay by FC. Our results demonstrate that Zoledronate 100 m M can induce apoptosis in B malignant lymphocytes despite previous treatment and MDR phenotype. So, patients previously treated with distinct therapeutic agents or not can potentially benefit from the anti-tumour effect of Zoledronate. This is the first work demonstrating the anti-tumour effect of Zoledronate in newly obtained cells from patients with B-CLL and LGL. These results in association with evidences from recent studies suggest that Zoledronate may have an important and complementary role in hematological malignant diseases.

scholarly journals Contrasting nucleolar activity in callus of beet and garlic as visualised by a new silver staining technique.

CYTOLOGIA ◽  
1989 ◽  
Vol 54 (3) ◽  
pp. 553-558
Author(s):  
S. A. Armstrong ◽  
B. V. Ford-Lloyd
Keyword(s):  
Silver Staining ◽  
Staining Technique ◽  
Nucleolar Activity ◽  
Silver Staining Technique

Location of focal silver staining at endothelial gaps in inflamed venules examined by scanning electron microscopy

1995 ◽  
Vol 269 (3) ◽  
pp. L403-L418 ◽  
Author(s):  
A. Hirata ◽  
P. Baluk ◽  
T. Fujiwara ◽  
D. M. McDonald
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Endothelial Cell ◽  
Silver Nitrate ◽  
Silver Staining ◽  
Cell Junctions ◽  
Luminal Surface ◽  
Silver Deposits ◽  
Endothelial Gaps ◽  
Scanning Electron

The century-old histological technique of silver nitrate staining has proven to be extremely useful for visualizing endothelial cell borders and localizing endothelial gaps, but the significance of the staining is still not fully understood. To gain some insight into what silver nitrate stains, we developed a method that enabled us to use scanning electron microscopy with backscattered and secondary electron imaging to examine silver staining at endothelial cell borders of venules of the rat tracheal mucosa. We found that in normal venules, silver lines followed the smooth contour of cell borders. However, 1 min after endothelial permeability was increased by substance P, cell borders were irregular and displaced from the silver lines by as much as 4.3 microns, and the lines were accompanied by three types of silver deposits. Most common (46% of total) were annulus-shaped silver deposits that surrounded endothelial gaps. These deposits averaged 1.5 microns in width, were positioned symmetrically across cell borders, and were located at a depth of 0.3 micron beneath the luminal surface. Many endothelial gaps were partitioned into multiple pores (mean, 2.4) by fingerlike processes of endothelial cells. Surprisingly, the gaps occupied only 5.4% of the total area of the silver deposits and constituted 0.15% of the luminal surface of the leaky postcapillary venules. A second type of silver deposit (19% of total) was positioned asymmetrically with respect to the cell border and marked sites where endothelial cell margins still overlapped but appeared to be vertically separated by obliquely oriented gaps. A third type marked gaps at three-cell junctions; these were no more abundant than deposits elsewhere around the cell perimeter, suggesting that three-cell junctions were not unusually leaky sites. We conclude that silver nitrate marks endothelial cell borders and outlines endothelial cell gaps by staining an element of intercellular junctions. The annular shape of many silver deposits around gaps suggests that junctional elements in the apposing cells are separated during gap formation but are still present at the gap perimeter.

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