scholarly journals Combined light and fluorescent microscopical imaging of nucleolar organizer regions and cellular rRNA as detected by fluorescent in situ hybridization

Cytometry ◽  
1995 ◽  
Vol 19 (2) ◽  
pp. 171-176 ◽  
Author(s):  
László Pajor ◽  
Thomas W. Honeyman
Keyword(s):  
In Situ Hybridization ◽  
Fluorescent In Situ Hybridization ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions

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.

Karyotype characterization of the lake sturgeon, Acipenser fulvescens (Rafinesque 1817) by chromosome banding and fluorescent in situ hybridization

Genome ◽  
2004 ◽  
Vol 47 (4) ◽  
pp. 742-746 ◽  
Author(s):  
Francesco Fontana ◽  
Ronald M Bruch ◽  
Fred P Binkowski ◽  
Massimo Lanfredi ◽  
Milvia Chicca ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Satellite Dna ◽  
Fluorescent In Situ Hybridization ◽  
Lake Sturgeon ◽  
Centromeric Heterochromatin ◽  
Telomeric Sequence ◽  
Acipenser Fulvescens ◽  
Nucleolar Organizer Regions ◽  
C Banding

A karyotype analysis using several staining techniques was carried out on the North American lake sturgeon, Acipenser fulvescens. The chromosome number was found to be 2n = 262 ± 6. A representative karyotype of 264 chromosomes was composed of 134 meta- and submetacentrics, 70 telo- and acrocentrics, and 60 microchromosomes. The constitutive heterochromatin, revealed by C banding, was localized in various positions on several chromosomes, including microchromosomes. The signals of fluorescent in situ hybridization (FISH) with a HindIII satellite DNA probe were visible as centromeric heterochromatin blocks on 48 chromosomes. The telomeric repeat (TTAGGG)n detected by FISH was localized at both ends of all chromosomes and two chromosomes were entirely marked. Fluorescent staining with GC-specific chromomycin A3 showed recognizable fluorescent regions, whereas a more uniform base composition was revealed by the AT-specific 4',6-diamidino-2-phenylindole (DAPI). After silver staining, the active nucleolar organizer regions (NORs) were detected on 12 chromosomes. FISH with the 5S probe showed four signals on four small chromosomes. Our data suggest that A. fulvescens is a tetraploid species.Key words: karyotype, C banding, telomeric sequence, fluorochrome staining, satellite DNA, 5S rDNA.

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.

Chromosomal and genomic organization of Ty1-copia-like retrotransposon sequences in the genus Avena

Genome ◽  
1996 ◽  
Vol 39 (2) ◽  
pp. 410-417 ◽  
Author(s):  
A. Katsiotis ◽  
T. Schmidt ◽  
J. S. Heslop-Harrison
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Sequence ◽  
Genomic Library ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
Reaction Products ◽  
D Genome ◽  
Avena Species ◽  
Copy Numbers

A cloned repetitive sequence, pAvKB30, obtained from an Avena vaviloviana (AB genome) genomic library, along with two polymerase chain reaction products derived from the conserved region of the reverse transcriptase (RT) gene of retrotransposons, were characterized molecularly and cytologically. The cloned DNA fragment was a dispersed repeat present in all Avena species used in this study (A. strigosa, A. clauda, A. vaviloviana, A. magna, and A. sativa). The fragment was sequenced (210 bp) and found to be 69.5% homologous to part of WIS-2-1A, and 60.5% homologous to the leader sequence of BARE-1; both of these elements have been characterized as Ty1-copia-like retrotransposons in wheat and barley, respectively. In situ hybridization of pAvKB30 to diploid, tetraploid, and hexaploid oat species revealed that the probe is present on both arms of all chromosomes (A, B, C, and D genomes) but is excluded from their centromeric and nucleolar organizer regions. By using double in situ hybridization in hexaploid A. sativa (ACD genome), pAvKB30 was found to be present in lower copy numbers in C-genome chromosomes compared with A- and D-genome chromosomes. Furthermore, under low stringency conditions, pAvKB30 hybridized on Southern blots containing barley, wheat, rye, and Arrhenatherum DNA. However, under high stringency conditions, it hybridized only on Arrhenatherum DNA, which is considered to be the genus most closely related to Avena. All Avena species included in this study yielded a PCR product when the primers from the RT domain of retrotransposons were used. Two products, rtA, obtained by using A. strigosa (As genome) as template, and rtC, obtained by using A. clauda (Cp genome) as template, gave Southern and in situ hybridization results similar to pAvKB30, but each was more abundant in its genome of origin. Key words : genomes, oats, in situ hybridization, translocations, repetitive sequence.

Cyto-evolution of Boronia genomes revealed by fluorescent in situ hybridization with rDNA probes

Genome ◽  
2003 ◽  
Vol 46 (3) ◽  
pp. 507-513 ◽  
Author(s):  
Fucheng Shan ◽  
Guijun Yan ◽  
Julie A Plummer
Keyword(s):  
Fluorescent In Situ Hybridization ◽  
Diploid Species ◽  
Nucleolar Organizer ◽  
Nucleolar Organizer Regions ◽  
26S Rdna ◽  
Rdna Sequences ◽  
Gene Copies ◽  
Rdna Sites ◽  
Rdna Copy

The physical location of the 25S–26S rDNA sequences was examined in 11 taxa of nine species of Boronia. In diploid species, two rDNA sites were detected in Boronia clavata (2n = 14), Boronia pinnata 'White' (2n = 22), and Boronia chartacea (2n = 32); four in Boronia megastigma (2n = 14) and Boronia denticulata (2n = 18); six in Boronia pinnata 'Pink' (2n = 22); and eight in Boronia molloyae (2n = 16). Eleven sites were found in Boronia heterophylla 'Red' and 'Near White' (2n = 15), but only two active nucleolar organizer regions (NORs) were observed. In polyploid species, Boronia pilosa (2n = 44) had four rDNA sites, while Boronia coerulescens (2n = 72) had six. Most of the rDNA sequences were terminal, but a few were interstitial. There were also differences in signal intensity indicating that the gene copies between and within rDNA sites might be different. The result suggests that considerable chromosome rearrangements have occurred during Boronia cyto-evolution, leading to variation among Boronia taxa in rDNA copy number, site number, and location. These changes together with dysploid reduction during cyto-evolution have made the Boronia genome considerably diverse in chromosome number, genome organization, and chromosome structure.Key words: physical mapping, FISH, chromosome, Rutaceae.

scholarly journals Mapping 18S ribosomal genes in fish of the genus Brycon (Characidae) by fluorescence in situ hybridization (FISH)

2000 ◽  
Vol 23 (1) ◽  
pp. 135-138 ◽  
Author(s):  
Adriane Pinto Wasko ◽  
Pedro Manoel Galetti Jr.
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Nucleolar Organizer ◽  
Rrna Genes ◽  
Nucleolar Organizer Regions ◽  
Single Chromosome ◽  
Individual Variations ◽  
18S Rrna Genes ◽  
High Level

The present study provides data on the nucleolar organizer regions (NORs) of seven Brycon species based on mapping of the 18S rRNA genes by fluorescence in situ hybridization (FISH). Fluorescent signals were observed on the telomere of the long arm of two large submetacentric chromosomes, thus confirming the number and location of NORs previously revealed by other classical cytogenetic techniques. Although there were no inter- or intra-individual variations in the number and location of the 18S loci, NOR size polymorphism was observed between homologous chromosomes. The clustering and conservation of NORs in a single chromosome pair indicates a high level of NOR stability among species of the genus Brycon.

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