Regularities of in situ nick translation labeling pattern of cereal metaphase chromosomes

Genome ◽  
1991 ◽  
Vol 34 (1) ◽  
pp. 131-138 ◽  
Author(s):  
M. V. Kost ◽  
N. L. Bolsheva ◽  
N. S. Badaev ◽  
A. V. Zelenin
Keyword(s):  
Active Regions ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Nick Translation ◽  
Active Chromatin ◽  
Metaphase Chromosomes ◽  
B Genome ◽  
Structural Conformation ◽  
Labeling Pattern

A method of nick translation in situ that makes it possible to detect regions hypersensitive to DNase I in metaphase chromosomes has been modified for plants. Such regions have a peculiar structural conformation typical of transcriptionally active chromatin. The distribution of transcriptionally active regions labeled with [3H]TTP along the length of individual chromosomes of rye and B genome of wheat was studied. The following regularities of this distribution were found: a weak labeling of heterochromatic regions; an increase in the intensity of labeling in regions located at the border of euchromatin and heterochromatin and in the nucleolar organizer regions. Each chromosome of the B and R genomes has its own labeling pattern, but some similarities in the labeling pattern in all chromosomes have been revealed. Such similarities are characteristic of homoeologous chromosomes.Key words: wheat, modified nick translation, rye.

DNA sequence mapping in interphase and metaphase chromosomes by fluorescence in situ hybridization

1992 ◽  
Vol 50 (1) ◽  
pp. 496-497
Author(s):  
Barbara Trask ◽  
Susan Allen ◽  
Anne Bergmann ◽  
Mari Christensen ◽  
Anne Fertitta ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequence ◽  
Dna Sequences ◽  
Dual Band ◽  
Nick Translation ◽  
Metaphase Chromosomes ◽  
Band Pass ◽  
Texas Red ◽  
Fluorescent Spot

Using fluorescence in situ hybridization (FISH), the positions of DNA sequences can be discretely marked with a fluorescent spot. The efficiency of marking DNA sequences of the size cloned in cosmids is 90-95%, and the fluorescent spots produced after FISH are ≈0.3 μm in diameter. Sites of two sequences can be distinguished using two-color FISH. Different reporter molecules, such as biotin or digoxigenin, are incorporated into DNA sequence probes by nick translation. These reporter molecules are labeled after hybridization with different fluorochromes, e.g., FITC and Texas Red. The development of dual band pass filters (Chromatechnology) allows these fluorochromes to be photographed simultaneously without registration shift.

Sequential silver staining and in situ hybridization reveal a direct association between rDNA levels and the expression of homologous nucleolar organizing regions: a hypothesis for NOR structure and function

1998 ◽  
Vol 111 (10) ◽  
pp. 1433-1439
Author(s):  
F. Zurita ◽  
R. Jimenez ◽  
M. Burgos ◽  
R.D. de la Guardia
Keyword(s):  
Transcription Factors ◽  
In Situ Hybridization ◽  
Silver Staining ◽  
Organizer Region ◽  
Nucleolar Organizer ◽  
Interindividual Variation ◽  
Nucleolar Organizer Regions ◽  
Single Pair ◽  
Ribosomal Cistrons

We have developed a procedure for sequential silver staining and in situ hybridization to analyze the relationship between the amount of rDNA present in nucleolar organizer regions, as estimated by in situ hybridization, and their level of expression, as estimated by the silver signal. For simplicity we used cells from the insectivorous mole Talpa occidentalis, which have a single pair of nucleolar organizer regions in chromosome pair 3. The relative content of ribosomal cistrons was also related to the hierarchy of activation of the nucleolar organizer regions present in this chromosomal pair. Statistical analyses demonstrated that both the relative level of expression and the activation hierarchy depended mainly on the number of ribosomal cistrons in nucleolar organizer regions. We propose a functional two-step hypothesis, which is consistent with most known data concerning interchromosomal, intercellular and interindividual variation in a number of plant and animal species, including Talpa occidentalis. In step one, the first available transcription factors bind randomly to the ribosomal promoters, such that larger nucleolar organizer regions are more likely to recruit them. In the second step the remaining transcription factors are recruited in a cooperative way, thus completing activation of one nucleolar organizer region, before the next one becomes active.

scholarly journals Different Proliferation Patterns in Breast Cancer: AgNOR Measurements in ER-Negative and ER-Positive Tumor Cells

2000 ◽  
Vol 20 (4) ◽  
pp. 155-162 ◽  
Author(s):  
Lukas Günther ◽  
Peter Hufnagl ◽  
Klaus‐Jürgen Winzer ◽  
Hans Guski
Keyword(s):  
Breast Cancer ◽  
Tumor Cells ◽  
Nucleolar Organizer ◽  
Growth Fraction ◽  
Nucleolar Organizer Regions ◽  
Ki 67 ◽  
Er Positive ◽  
Negative Tumor ◽  
Invasive Breast Carcinomas

The relation between estrogen receptors (ER) and argyrophilic nucleolar organizer regions (AgNORs)in situwithin human breast cancer cells was analyzed. For AgNOR measurements in 49 invasive breast carcinomas, a new reproducible staining method for dual demonstration of ER and AgNORs was applied. Quantitative AgNOR variables were determined in ER‐positive and ER‐negative tumor cells by digital image analysis. The relationships between AgNOR parameters of ER‐positive and ER‐negative cells and other prognostic factors of breast cancer [Bloom–Richardson‐Grading and growth fraction (Ki‐67 index)] were investigated. A higher AgNOR content in ER‐negative cells and a special clustering phenomenon in ER‐positive tumor cells were found. Correlation with other criteria of malignant potential could be exclusively demonstrated for ER‐negative cells. ER‐negative cells of breast cancer can be characterized as the more malignant and possibly prognosis‐dictating cell fraction. Thus, ER‐negative cells probably contribute more to the progression of the tumor disease and furthermore to the prognosis than ER‐positive cells. We recommend measurement AgNORs exclusively in ER‐negative cells of breast cancer.

Comparative cytogenetic analysis of Avena macrostachya and diploid C-genome Avena species

Genome ◽  
2010 ◽  
Vol 53 (2) ◽  
pp. 125-137 ◽  
Author(s):  
Ekaterina D. Badaeva ◽  
Olga Yu. Shelukhina ◽  
Axel Diederichsen ◽  
Igor G. Loskutov ◽  
Vitaly A. Pukhalskiy
Keyword(s):  
5S Rdna ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Rrna Gene ◽  
Avena Species ◽  
Rdna Sites ◽  
C Genome ◽  
Genome Group ◽  
26S Rrna

The chromosome set of Avena macrostachya Balansa ex Coss. et Durieu was analyzed using C-banding and fluorescence in situ hybridization with 5S and 18S-5.8S-26S rRNA gene probes, and the results were compared with the C-genome diploid Avena L. species. The location of major nucleolar organizer regions and 5S rDNA sites on different chromosomes confirmed the affiliation of A. macrostachya with the C-genome group. However, the symmetric karyotype, the absence of “diffuse heterochromatin”, and the location of large C-band complexes in proximal chromosome regions pointed to an isolated position of A. macrostachya from other Avena species. Based on the distribution of rDNA loci on the C-genome chromosomes of diploid and polyploid Avena species, we propose a model of the chromosome alterations that occurred during the evolution of oat species.

Dynamic Changes in the Distribution of a Satellite Homologous to Intergenic 26-18S rDNA Spacer in the Evolution of Nicotiana

Genetics ◽  
2004 ◽  
Vol 166 (4) ◽  
pp. 1935-1946
Author(s):  
K Y Lim ◽  
K Skalicka ◽  
B Koukalova ◽  
R A Volkov ◽  
R Matyasek ◽  
...  
Keyword(s):  
18S Rdna ◽  
Intergenic Spacer ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Dynamic Nature ◽  
Dynamic Changes ◽  
Nicotiana Tomentosiformis ◽  
Copy Numbers ◽  
Location Patterns

Abstract An ∼135-bp sequence called the A1/A2 repeat was isolated from the transcribed region of the 26-18S rDNA intergenic spacer (IGS) of Nicotiana tomentosiformis. Fluorescence in situ hybridization (FISH) and Southern blot analysis revealed its occurrence as an independent satellite (termed an A1/A2 satellite) outside of rDNA loci in species of Nicotiana section Tomentosae. The chromosomal location, patterns of genomic dispersion, and copy numbers of its tandemly arranged units varied between the species. In more distantly related Nicotiana species the A1/A2 repeats were found only at the nucleolar organizer regions (NOR). There was a trend toward the elimination of the A1/A2 satellite in N. tabacum (tobacco), an allotetraploid with parents closely related to the diploids N. sylvestris and N. tomentosiformis. This process may have already commenced in an S3 generation of synthetic tobacco. Cytosine residues in the IGS were significantly hypomethylated compared with the A1/A2 satellite. There was no clear separation between the IGS and satellite fractions in sequence analysis of individual clones and we found no evidence for CG suppression. Taken together the data indicate a dynamic nature of the A1/A2 repeats in Nicotiana genomes, with evidence for recurrent integration, copy number expansions, and contractions.

NUCLEOLAR ORGANIZER REGIONS IN THE RABBIT (ORYCTOLAGUS CUNICULUS) AS SHOWN BY SILVER STAINING

1978 ◽  
Vol 20 (3) ◽  
pp. 377-382 ◽  
Author(s):  
Patricia A. Martin-Deleon ◽  
Dorene L. Petrosky ◽  
M. Eileen Fleming
Keyword(s):  
New Zealand ◽  
Silver Staining ◽  
Oryctolagus Cuniculus ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Telomeric Region ◽  
Metaphase Chromosomes ◽  
Domestic Rabbit ◽  
Silver Precipitation ◽  
Secondary Constrictions

Nucleolar organizer regions (NOR's) were demonstrated in metaphase chromosomes of the domestic rabbit, Oryctolagus cuniculus (L.) (New Zealand white strain) using silver staining. Sequential quinacrine banding and a modification of the Ag-AS silver precipitation technique with duplicate photography allowed identification of silver staining NOR's on the short arms of chromosomes 13, 16, and 20, as well as the telomeric region of the long arms of number 21 in some cells. Chromosomes 13, 16 and 20 all have subterminal to terminal centromeres, often showed satellites and secondary constrictions, and were sometimes involved in associations.

scholarly journals Cytogenetic analyses in Trinomys (Echimyidae, Rodentia), with description of new karyotypes

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5316 ◽  
Author(s):  
Naiara Pereira Araújo ◽  
Cayo Augusto Rocha Dias ◽  
Rodolfo Stumpp ◽  
Marta Svartman
Keyword(s):  
Chromosome Mapping ◽  
Nucleolar Organizer ◽  
Interspecific Variation ◽  
Nucleolar Organizer Regions ◽  
45S Rdna ◽  
Banding Patterns ◽  
Pericentric Inversions ◽  
Cytogenetic Analyses ◽  
First Time

Trinomys Thomas (1921) is a terrestrial genus of spiny rats endemic to the Brazilian areas of Atlantic Forest and the transitional areas of Cerrado and Caatinga. Although most species have been already karyotyped, the available cytogenetic information is mostly restricted to diploid and fundamental numbers. We analyzed the chromosomes of two Trinomys species: Trinomys moojeni (2n = 56, FN = 106) and Trinomys setosus setosus (2n = 56, FN = 106 and 2n = 56, FN = 108). Our analyses included GTG- and CBG-banding, silver-staining of the nucleolar organizer regions, and chromosome mapping of telomeres and 45S rDNA by fluorescent in situ hybridization (FISH). Comparative GTG- and CBG-banding suggested that the interspecific variation may be due to rearrangements such as pericentric inversions, centromere repositioning, and heterochromatin variation. We report two new karyotypes for T. s. setosus and describe for the first time the banding patterns of the two Trinomys species.

scholarly journals Cytogenetics of two sympatric Corydoras species (pisces, siluriformes, challichtyidae) of Southern Brazil

2006 ◽  
Vol 66 (1b) ◽  
pp. 191-198 ◽  
Author(s):  
R. F. Artoni ◽  
M. L. Terêncio ◽  
M. R. Vicari ◽  
M. C. A. Matiello ◽  
M. M. Cestari ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Diploid Number ◽  
Constitutive Heterochromatin ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Great Similarity ◽  
Southern Brazil ◽  
Rdna Probe ◽  
Karyotypic Formula

Karyotypic data are presented for two sympatric Corydoras species of the Lagoa Dourada, namely, C. ehrhadti and C. paleatus, which are found in the upper Tibagi river basin (Ponta Grossa, State of Paraná, Brazil). The same diploid number and karyotypic formula were observed in both species/populations. A great similarity in the constitutive heterochromatin distribution and in the activity of nucleolar organizer regions was also found. The use of in situ hybridization with a fluorescent 18S rDNA probe allowed for the identification of the species/populations through the location of ribosomal sites.

scholarly journals Cytogenetic analyses of two Curimatidae species (Pisces; Characiformes) from the Paranapanema and Tietê Rivers

2007 ◽  
Vol 67 (2) ◽  
pp. 333-338 ◽  
Author(s):  
LVS De Rosa ◽  
F. Foresti ◽  
C. Martins ◽  
C. Oliveira ◽  
PE. Sobrinho ◽  
...  
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Constitutive Heterochromatin ◽  
Nucleolar Organizer ◽  
Terminal Region ◽  
Nucleolar Organizer Regions ◽  
Micro Structure ◽  
Isolated Populations ◽  
Cytogenetic Characterization ◽  
Cytogenetic Analyses

Cytogenetic analyses were performed in two Curimatidae species (Steindachnerina insculpta and Cyphocharax modesta) from the Paranapanema and Tietê Rivers (São Paulo State, Brazil), showing a karyotype composed of 54 meta-submetacentric chromosomes in both species. Silver- and chromomycyn-staining and fluorescent in situ hybridization (FISH) using a 18S rDNA probe indicated that the nucleolar organizer regions (NORs) of both species are localized in the terminal region of the long arm of two metacentric chromosomes. Although a single NOR system was evidenced in both analyzed species, S. insculpta and C. modesta presented the nucleolar organizer regions in distinct chromosome pairs, indicating that these cistrons can be considered cytogenetic markers. Variation on the amount and distribution of the constitutive heterochromatin (C-bands) could also be detected between the two species - while S. insculpta presented few heterochromatic blocks, intensely stained C-bands were evidenced in C. modesta specially in the terminal region of the long arm of the NOR-bearing chromosomes. Although most Curimatidae species have been characterized by homogeneous karyotypes, isolated populations could be established under different environmental conditions leading to karyotype micro-structure variations specially related to the NORs localization and C-banding distribution. The obtained data were useful for the cytogenetic characterization and differentiation of S. insculpta and C. modesta and could be used in evolutionary inferences in the Curimatidae group.

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