Silver staining of nucleolar organizer region associated proteins using polyethylene glycol as the protective colloidal developer

1990 ◽  
Vol 22 (10) ◽  
pp. 555-559 ◽  
Author(s):  
D. C. Rowlands ◽  
J. Crocker ◽  
J. G. Ayres
2012 ◽  
Vol 10 (1) ◽  
pp. 34-39 ◽  
Author(s):  
S Karki ◽  
A Jha ◽  
G Sayami

Background Serous effusion smears reported as “suspicious for malignancy” pose problems in clinical management. Silver staining for argyrophilic nucleolar organizer regions (AgNOR) has proved useful in making a cytopathologic differential diagnosis between benign and malignant cells. Nucleolar organizer regions(NORs) are loops of DNA located in acrocentric chromosomes. These NORs are visualized by silver staining technique that recognizes these argyrophilia associated proteins which are increased in malignancy. Objective This study aimed to distinguish reactive mesothelial cells from malignant cells in serous effusions using these NORs. Methods A total of 174 serous effusions received at the Department of Pathology, TUTH, during a period of one year were included in the study. Smears were studied by conventional Papanicolaou and Giemsa stains. AgNOR counts, variation in size and dispersion of AgNOR dots in smears were graded and compared in malignant and non-malignant effusions. Results Mean AgNOR counts of 10.43±0.73 and 10.21±0.51 in malignant peritoneal and pleural effusions, respectively, were significantly (p<0.0001) greater as compared with counts of 2.12±0.54 and 2.11±0.54 in non-malignant effusions. The AgNORs were irregular in shape in malignant effusions whereas they were comparatively larger, single dots in benign effusions. AgNOR size and dispersion were of higher grade in significantly greater proportion of malignant as compared with non malignant effusions (p<0.0001). Of the cytologically suspicious samples, nine were in the malignant range and one was in the benign range. Conclusion AgNOR study appears to be clinically useful as an additional diagnostic tool for use in serous effusion when the cytologic diagnosis is difficult. KATHMANDU UNIVERSITY MEDICAL JOURNAL  VOL.10 | NO. 1 | ISSUE 37 | JAN - MAR 2012 | 44-47 DOI: http://dx.doi.org/10.3126/kumj.v10i1.6913


1993 ◽  
Vol 41 (9) ◽  
pp. 1413-1417 ◽  
Author(s):  
G Méhes ◽  
E Kálmán ◽  
L Pajor

Nucleolar organizer regions (NORs) are nucleolus-forming rDNA loops associated with argyrophil proteins, the amount of which varies according to the proliferative state of the cell. It has been presumed that the nucleolar protein-related thiol groups may have a role in selective silver staining. We investigated the nuclear thiol distribution with a fluorescent thiol reagent, coumarinyl-phenyl-maleimide (CPM) in human K-562 myeloblast cultures and found that SH group-related fluorescence was brightest in the area of nucleoli, which became highly selective after RNAse digestion. A remarkable co-localization of AgNOR silver reaction and CPM fluorescence was observed, although occupation of the SH groups by CPM did not prevent the silver staining. We applied the stain to dual-parameter flow cytometry in combination with DNA content measurements, which provide further information on nucleolar function and changes in experimental and pathological specimens.


2002 ◽  
Vol 115 (14) ◽  
pp. 2839-2846
Author(s):  
Ana D. Caperta ◽  
Nuno Neves ◽  
Leonor Morais-Cecílio ◽  
Rui Malhó ◽  
Wanda Viegas

The standard rye cultivar `Imperial' and a structural variant carrying an intact 1R chromosome and two telocentric 1R chromosomes (short and long arms)were used to investigate expression patterns of homologous rDNA loci, and the influence of chromosome structural change on their interphase organisation and relative disposition. Sequential silver staining and in situ hybridization with the rDNA probe pTa71, established a correspondence between the expression and organization patterns of rDNA domains in metaphase and interphase cells. In most cells of the cultivar Imperial, nucleolar organizer region (NOR)silver staining on metaphase chromosomes with equivalent numbers of rDNA genes revealed a size heteromorphism between homologous rDNA loci, resulting from their differential expression. NOR heteromorphism in the structural variant line was significantly reduced. The preferential activity of one NOR over its homologue was found to be random within cells and independent of parental origin. Nucleotypic modifications mediated by changes in the 1R chromosome structure include increased proximity between homologous rDNA loci in interphase, and an increase in the frequency of cells with intra-nucleolar ribosomal condensed chromatin. These results seem to indicate a `sequence recognition' process for the regulation of homologous loci.


1985 ◽  
Vol 33 (5) ◽  
pp. 389-399 ◽  
Author(s):  
F J Moreno ◽  
D Hernandez-Verdun ◽  
C Masson ◽  
M Bouteille

Nucleolar organizer region (NOR) silver staining was applied to sections of fixed material. A positive reaction on cryo-ultrathin sections was found as well as on semithin and ultrathin Lowicryl sections. Repeatable staining that was easy to control was obtained by a one-step procedure after aldehyde-Carnoy fixation. Fixation of the material by formaldehyde and glutaraldehyde alone in cacodylate buffer also maintained reaction selectivity when ammonium chloride was used after fixation. Enzymatic digestion by pronase, RNase A, DNase I, or micrococcal nuclease was applied to ultrathin Lowicryl sections. Pronase digestion removed the silver-stained proteins, whereas digestion by the nucleases did not. A routine procedure is proposed for easy NOR silver staining of sections that preserves a good tissue ultrastructure and is also compatible with cytochemical and immunological investigations.


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