NOR Chromosome Research Articles

B Chromosomes and Cytogenetic Characteristics of the Common Nase Chondrostoma nasus (Linnaeus, 1758).

Supernumerary B chromosomes (Bs) are very promising structures, among others, in that they are an additional genomic compartment for evolution. In this study, we tested the presence and frequency of B chromosomes and performed the first cytogenetic examination of the common nase (Chondrostoma nasus). We investigated the individuals from two populations in the Vistula River basin, in Poland, according to the chromosomal distribution of the C-bands and silver nucleolar organizer regions (Ag-NORs), using sequential staining with AgNO3 and chromomycin A3 (CMA3). Furthermore, we analyzed the chromosomal localization of two rDNA families (45S and 5S rDNA) using fluorescence in situ hybridization (FISH) with rDNA probes. Chondrostoma nasus individuals showed a standard (A) chromosome set consisting of 2n = 50: 12 metacentric, 32 submetacentric, and 6 acrocentric chromosomes (NF = 94). Fourteen out of the 20 analyzed individuals showed 1–2 mitotically unstable submetacentric B chromosomes of different sizes. Six of them, in 14.1% of the analyzed metaphase plates, had a single, medium-sized submetacentric B (Bsm) chromosome (2n = 51) with a heterochromatic block located in its pericentromeric region. The other seven individuals possessed a Bsm (2n = 51) in 19.4% of the analyzed metaphase plates, and a second Bsm chromosome (2n = 52), the smallest in the set, in 15.5% of metaphase plates, whereas one female was characterized by both Bsm chromosomes (2n = 52) in 14.3% of the analyzed metaphase plates. AgNORs, GC-rich DNA sites, and 28S rDNA hybridization sites were observed in the short arms of two submetacentric chromosome pairs of A set. The constitutive heterochromatin was visible as C bands in the centromeric regions of almost all Chondrostoma nasus chromosomes and in the pericentromeric region of several chromosome pairs. Two 5S rDNA hybridization sites in the pericentromeric position of the largest acrocentric chromosome pair were observed, whereas two other such sites in co-localization on a smaller pair of NOR chromosomes indicate a species-specific character. The results herein broaden our knowledge in the field of B chromosome distribution and molecular cytogenetics of Chondrostoma nasus: a freshwater species from the Leuciscidae family.

Anadolu’da Yaşayan Endemik Üç Pseudophoxinus Bleeker, 1860 (Teleostei: Leuciscidae) Türünün CMA3 ve DAPI Boyalı Kromozomları

The karyotypes and other chromosomal markers of representatives of the genus Pseudophoxinus have been little studied. Therefore, this study documents chromosomal NOR phenotypes as revealed using Chromomycin A3 (CMA3) and DAPI chromosomal stainings in three species Pseudophoxinus elizavetae Bogutskaya, Küçük & Atalay, 2007, P. firati Bogutskaya, Küçük & Atalay, 2007 and P. hittitorum Freyhof & Özuluğ, 2010, endemic in Anatolia. Simple NOR phenotype with only two CMA3 positive signals were observed in the karyotype of P. firati whereas derived NOR phenotype with four CMA3 positive signals were observed in those of P. elizavetae and P. hittitorum, respectively. No DAPI positive signal was detected in karyotypes in all three species. This study described new NOR phenotypes, i.e. number and position of major rDNA genes, in genomes of three Pseudophoxinus species contributing thus to known diversity of NOR phenotypes in otherwise karyotypically highly conservative leuciscid fishes.

First Study on Chromosomal Feature and NORs Localization of the Yellow-spotted Keelback Snake, Fowlea flavipunctatus (Squamata, Natricinae) in Thailand

First Study on Chromosomal Feature and NORs Localization of the Yellow-spotted Keelback Snake, Fowlea flavipunctatus (Squamata, Natricinae) in Thailand

The karyotypes of five species of the Scinax perpusillus group (Amphibia, Anura, Hylidae) of southeastern Brazil show high levels of chromosomal stabilization in this taxon.

Based on morphological, bioacoustics, and morphological traits, the genus Scinax has been subdivided into two major clades: S. catharinae and S. ruber. The first clade includes S. catharinae and S. perpusillus groups, whereas the second clade includes S. rostratus and S. uruguayus groups. Chromosome morphology, NOR and C-banding patterns of variation support these clades. This study aims the cytogenetic characterization of five species currently included in the S. perpusillus group: Scinax sp. (gr. perpusillus), S. arduous, S. belloni, S. cosenzai, and S. v-signatus, including standard cytogenetic techniques and repetitive DNA FISH probes. All species had 2n=24 chromosomes. Nucleolar organizing regions occurred in chromosome pair 6 in all species, but differed in their locations among some species, suggesting a putative synaponomastic character for the clade. In S. belloni, the first chromosome pair was a metacentric, contrasting with the submetacentric first pair reported in all other species of the genus. Scinax sp. (gr. perpusillus) and S. v-signatus had similar karyotypic formulae, suggesting they are related species. Scinax cosenzai had a divergent C-banding pattern. Repetitive DNA probes hybridized more frequently in chromosomal subtelomeric regions in all species indicating recent cladogenesis in these species. Karyotypic evidence indicates unreported high levels of stabilization within S. perpusillus and in S. catharinae clade, resulting in a wealth of characters potentially informative for higher phylogenetic analyses.

First Report of NOR Polymorphism and Chromosome Analysis of John's Snapper, <i>Lutjanus johnii</i> (Perciformes, Lutjanidae) in Thailand

First Report of NOR Polymorphism and Chromosome Analysis of John's Snapper, <i>Lutjanus johnii</i> (Perciformes, Lutjanidae) in Thailand

Karyotype and NOR-banding of mitotic chromosomes of some Vitis L. species

Chromosome studies were performed in V. champinii, V. cinerea, V. girdiana, V. labrusca, V. rotundifolia, V. rupestris and V. vinifera with the purpose of species characterization using chromosome morphometric data and NOR banding. A median ideogram was obtained for each species. The karyotype formula obtained varied from 7m + 12sm to 9m + 11sm. The species showed moderate chromosome asymmetry values according to TF% form, Stebbins, Romero Zarco and Paszko indices. V. champinii and V. girdiana were apart from the other species by CVcl and CVci graphic representation and also formed a group apart in the dendrogram based on Euclidian distances. The chromosome pair number 3 harbors the secondary constriction and a satellite segment in all species analyzed with Giemsa staining and it may be the same observed after NOR banding technique. It seems that the process of speciation in the North American Euvitis species studied involved some discrete changes in chromosome morphometry which have been reflected in the asymmetry index.

FISH in analysis of gamma ray-induced micronuclei formation in barley

A micronucleus test in combination with fluorescent in situ hybridization (FISH) using telomere-, centromere-specific probes and 5S and 25S rDNA was used for a detailed analysis of the effects of gamma ray irradiation on the root tip meristem cells of barley, Hordeum vulgare (2n = 14). FISH with four DNA probes was used to examine the involvement of specific chromosomes or chromosome fragments in gamma ray-induced micronuclei formation and then to explain their origin. Additionally, a comparison of the possible origin of the micronuclei induced by physical and chemical treatment: maleic hydrazide (MH) and N-nitroso-N-methylurea (MNU) was done. The micronuclei induced by gamma ray could originate from acentric fragments after chromosome breakage or from whole lagging chromosomes as a result of a dysfunction of the mitotic apparatus. No micronuclei containing only centromeric signals were found. An application of rDNA as probes allowed it to be stated that 5S rDNA–bearing chromosomes are involved in micronuclei formation more often than NOR chromosomes. This work allowed the origin of physically- and chemically-induced micronuclei in barley cells to be compared: the origin of micronuclei was most often from terminal fragments. FISH confirmed its usefulness in the characterization of micronuclei content, as well as in understanding and comparing the mechanisms of the actions of mutagens applied in plant genotoxicity.

Chromosome phylogeny of the subfamily Pitheciinae (Platyrrhini, Primates) by classic cytogenetics and chromosome painting.

BackgroundThe New World monkey (Platyrrhini) subfamily Pitheciinae is represented by the genera Pithecia, Chiropotes and Cacajao. In this work we studied the karyotypes of Pithecia irrorata (2n = 48) and Cacajao calvus rubicundus (2n = 45 in males and 2n = 46 in females) by G- and C-banding, NOR staining and chromosome painting using human and Saguinus oedipus whole chromosome probes. The karyotypes of both species were compared with each other and with Chiropotes utahicki (2n = 54) from the literature.ResultsOur results show that members of the Pitheciinae have conserved several chromosome forms found in the inferred ancestral Platyrrhini karyotype (associations of human homologous segments 3a/21, 5/7a, 2b/16b, 8a/18, 14/15a and 10a/16a). Further, the monophyly of this subfamily is supported by three chromosomal synapomorphies (2a/10b, an acrocentric 15/14 and an acrocentric human 19 homolog). In addition, each species presents several autapomorphies. From this data set we established a chromosomal phylogeny of Pitheciinae, resulting in a single most parsimonious tree.ConclusionsIn our chromosomal phylogeny, the genus Pithecia occurred in a more basal position close to the inferred ancestor of Platyrrhini, while C. c. rubicundus and C. utahicki are closely related and are linked by exclusive synapomorphies.

Karyological Observations in Medicago Section Dendrotelis (Fabaceae)

Medicago section Dendrotelis comprises polyploid, woody shrubs restricted to rocky and cliff faces in coastal places of the Mediterranean basin, showing a very close morphology. This has resulted in controversial taxonomic studies, differing in the circumscription and number of recognized species (from one to three). In the present study, cytogenetic studies (meiotic analysis, C-banding, Ag-NOR banding) were made in M. arborea, M. strasseri, and M. citrina to determine their karyological and evolutionary relationships. The observation of meiocytes revealed a regular meiotic cycle in all species. At pachytene stage, chromosomes showed constitutive heterochromatic blocks (knobs) that were species-specific, and clearly discriminate M. arborea and M. strasseri from M. citrina. Conventional karyotypes of M. strasseri and M. arborea were similar concerning chromosome classes, the apparent chromosome length, and the morphology of the single NOR chromosome pair. However, C-banding technique revealed karyotypic differences between both species. In contrast, M. citrina exhibited a slightly asymmetric karyotype, the presence of two pairs of chromosomes showing secondary constrictions, a similar apparent chromosome length, and two pairs of active rDNA loci. Its C-banded karyotype strikingly differed from those observed in M. arborea and M. strasseri, and only three chromosome pairs showed heterochromatic blocks. These cytogenetic data indicate a clear evolutionary split in woody medics (tetraploid vs hexaploid species), reflecting divergent patterns of karyological evolution. Taken together, all karyological data unequivocally support the recognition of M. citrina as a distinct species.

Quantitative analysis of NOR expression in a B chromosome of the grasshopper Eyprepocnemis plorans

The B24 chromosome in the Torrox population of the grasshopper Eyprepocnemis plorans is recurrently attached to a nucleolus in diplotene cells, indicating the activity of its distally located ribosomal DNA (rDNA). The frequency of males expressing the B chromosome nucleolus organizer region (B-NOR) almost doubled in 4 years. The likelihood of expressing the B-NOR increased with the B number and, in males expressing it, about 20% of their cells showed a nucleolus attached to the B. When active, the B-NOR contributed more than 25% of total cell nucleolar area (NA). Within males expressing the B-NOR, total cell NA did not differ between cells showing the active or inactive B-NOR, suggesting that total cell NA is tightly regulated in this species. However, this parameter tended to increase in this population from 1999 to 2004, in parallel to the neutralization process which is taking place in this population. Finally, an analysis of A chromosome NOR interdependence for activity revealed a positive correlation among autosomes but a negative correlation between autosomes and the X chromosome, the manifestation of which depends on B-NOR activity. These results are discussed in the context of the nucleolus as a sensor of the stress caused by parasitic B chromosomes.

Cytogenetic studies in North American minnows (Cyprinidae): Chromosomal NOR phenotypes of 12 species, with comments on cytosystematic relationships among 50 species

Chromosomal NOR phenotypes are documented for twelve species of North American cyprinid fishes whose NOR karyotypes have not been previously reported. Patterns of interspecific chromosomal NOR variation are then summarized for the fifty species of North American cyprinids surveyed to date. The degree of interspecific NOR variation among North American cyprinids suggests that the rate of chromosomal evolution within the group may have been higher than previously believed. The interspecific variants in chromosomal NORs are used as taxonomic and systematic characters in order to infer phylogenetic hypotheses of species relationships. The phylogenetic inferences drawn from the chromosomal NOR data provide a framework upon which future cytosystematic studies in North American cyprinids can be based.

Relationships of the Woody Medicago Species (Section Dendrotelis) Assessed by Molecular Cytogenetic Analyses

The organization of rDNA genes in the woody medic species from the agronomically important Medicago section Dendrotelis was analysed to gain insight into their taxonomic relationships, to assess the levels of infraspecific variation concerning ribosomal loci in a restricted and fragmented insular species (M. citrina) and to assess the nature of its polyploidy. Fluorescence in situ hybridization (FISH) was used for physical mapping of 5S and 45S ribosomal DNA genes in the three species of section Dendrotelis (M. arborea, M. citrina, M. strasseri) and the related M. marina from section Medicago. Genomic in situ hybridization (GISH) was used to assess the genomic relationships of the polyploid M. citrina with the putatively related species from section Dendrotelis. The diploid (2n = 16) M. marina has a single 45S and two 5S rDNA loci, a pattern usually detected in previous studies of Medicago diploid species. However, polyploid species from section Dendrotelis depart from expectations. The tetraploid species (2n = 32) M. arborea and M. strasseri have one 45S rDNA locus and two 5S rDNA loci, whereas in the hexaploid (2n = 48) M. citrina four 45S rDNA and five 5S rDNA loci have been detected. No single chromosome of M. citrina was uniformly labelled after using genomic probes from M. arborea and M. strasseri. Instead, cross-hybridization signals in M. citrina were restricted to terminal chromosome arms and NOR regions. FISH results support the close taxonomic interrelationship between M. arborea and M. strasseri. In these tetraploid species, NOR loci have experienced a diploidization event through physical loss of sequences, a cytogenetic feature so far not reported in other species of the genus. The high number of rDNA loci and GISH results support the specific status for the hexaploid M. citrina, and it is suggested that this species is not an autopolyploid derivative of M. arborea or M. strasseri. Further, molecular cytogenetic data do not suggest the hypothesis that M. arborea and M. strasseri were involved in the origin of M. citrina. FISH mapping can be used as an efficient tool to determine the genomic contribution of M. citrina in somatic hybrids with other medic species.

Cytogenetic studies in fishes of the genera Hassar, Platydoras and Opsodoras (Doradidae, Siluriformes) from Jarí and Xingú Rivers, Brazil

We studied the karyotypes of Hassar cf. orestis and an undescribed Hassar species from the Jari River and Opsodoras ternetzi, H. orestis and Platydoras cf. costatus from the Xingu River, all with 2n = 58. Constitutive heterochromatin is located in the centromere in most metacentric pairs; in some chromosomes this banding is not present, or it is located on the whole chromosome arm or in the distal regions. The NOR is located on a single biarmed pair at a distal region of the short arm in H. cf. orestis, H. orestis and P. cf. costatus at a distal region of the long arm in O. ternetzi and at a proximal region of the long arm in the Hassar species. In all species (except for Hassar sp.) the CMA3 analysis revealed a rich G-C region coincident with the NOR. Probably inversions occurred in the NOR chromosome during the chromosomal differentiation of the Doradidae species here described.

Isolation, cloning and characterization of two major satellite DNA families of rabbit ( Oryctolagus cuniculus)

We report here the isolation, cloning and characterization of two abundant centromeric satellite sequences (Rsat I and Rsat II) what are not related to each other, and that of a divergent subfamily (Rsat IIE) of rabbit ( Oryctolagus cuniculus). The Rsat I monomers had a 375 base pair (bp) average length, while repeat units Rsat II and Rsat IIE were ∼585 bp long. Variable amounts of Rsat I were detected by FISH at the centromeric region of 11 chromosome pairs of the complement. Rsat II hybridized to the centromere of 12 different chromosomes, and two of these were labeled also with the Rsat IIE probe. Two-color in situ hybridizations with the satellite probes and rDNA revealed that the NOR chromosomes carried different satellites. Rsat I was abundant on chromosome 20 and 21, but it was undetectable on chromosomes 13 and 16. Large Rsat II arrays were found on chromosomes 16, 20 and 21, but reduced amount was detected on chromosome 13. The variant Rsat IIE was prominent on chromosome 16, but was absent from the other rDNA-bearing chromosomes. The rDNA signal on chromosome 21 was localized to the 21q(ter) region, what can be a useful cytological marker in comparative cytological studies. These data show that rabbit chromosomes form at least four distinct groups based on the satellite composition of their centromeres. The differences in the chromosomal distribution of satellite families will help easy FISH identification of individual chromosomes, as well as to unveil the evolutionary history of the Leporidae karyotype.

Molecular cytogenetic definition of the chicken genome: the first complete avian karyotype.

Chicken genome mapping is important for a range of scientific disciplines. The ability to distinguish chromosomes of the chicken and other birds is thus a priority. Here we describe the molecular cytogenetic characterization of each chicken chromosome using chromosome painting and mapping of individual clones by FISH. Where possible, we have assigned the chromosomes to known linkage groups. We propose, on the basis of size, that the NOR chromosome is approximately the size of chromosome 22; however, we suggest that its original assignment of 16 should be retained. We also suggest a definitive chromosome classification system and propose that the probes developed here will find wide utility in the fields of developmental biology, DT40 studies, agriculture, vertebrate genome organization, and comparative mapping of avian species.

Chromosome Relationships between Anostomidae and Chilodontidae Fish (Characiformes).

Although many chromosome studies have been conduced in Anostomidae, the data are still concentrated in some genera. Other Characiformes families, such as Chilodontidae, are still cytogenetically unknown. In order to contribute to the cytogenetical knowledge on Anostomidae and Chilodontidae, the mitotic chromosomes of 3 anostomid species, Abramites solarii, Anostomus ternetzi and Pseudanos trimaculatus, and 2 chilodontids, Caenotropus labirynthicus and Chilodus punctatus, were analyzed using Giemsa staining, C-banding and nucleolus organizer region distribution detected by silver nitrate (Ag-NOR) and mithramycin (MM) staining. Although 54 biarmed chromosomes and 1 chromosome pair bearing NOR sites represent a general trait, heterochromatin distribution and NOR chromosome pair position are variable among Anostomidae and Chilodontidae species. This trend suggests that cryptic changes along the chromosome complement of these fishes have occurred, even thought the gross karyotype structure has been strongly conserved during their chromosome evolution. Furthermore, it appears to have a parallelism between morphological/ecological diversity and chromosomal evolution rate among anostomids and chilodontids.

Chromosome characterization of Trichotnycterus spegazzini (Siluriformes, Trichomycteridae) from three hydrographic basins of the Northwest of Argentina

Abstract Cytogenetic studies involving conventional air drying techniques, Giemsa staining,constitutive heterochromatin analysis and silver staining were carried out on T. spegazzini from three hydrographic basins of the northwest of Argentina. In spite of colour and subtle morphological differencies the three populations showed the same karyologycal formulae, 54 biarmed chromosomes, 42 M and 12 SM pairs, NF = 108, with a very conspicuous first metacentric pair. The NORs were located at the terminal region on the short arm on a single submetacentric medium sized pair. The C banding analysis showed the presence of small blocks in pericentromeric position in an single metacentric large chromosome pair (3° o 4°). The species showed a conservative karyotype, with plesiomorphic condition for chromosome number and NOR bands.

NOR polymorphism in the Iberian species Chondrostoma lusitanicum (Pisces: Cyprinidae)--re-examination by FISH.

Chromosomal polymorphism regarding the number of chromosomal NOR sites in the cyprinid fish Chondrostoma lusitanicum reported previously (Rodrigues & Collares-Pereira, 1996) was re-examined using fluorescence in situ hybridization (FISH) with a ribosomal DNA (rDNA) probe. All positive CMA3-bands contained ribosomal DNA documented by either two or four FISH positive signals in the respective karyotypes. This polymorphism suggests the occurrence of structural rearrangements of translocation type in rDNA region from one ancestral NOR-bearing chromosome pair ubiquitous among leuciscine cyprinid fishes to another pair. The absence of individuals heterozygous for this polymorphism is discussed.

Subcongenic Analysis of theIdd13Locus in NOD/Lt Mice: Evidence for Several Susceptibility Genes Including a Possible Diabetogenic Role for β2-Microglobulin

AbstractAlthough they share ∼88% of their genome with NOD mice including the H2g7 haplotype, NOR mice remain free of T cell-mediated autoimmune diabetes (IDDM), due to non-MHC genes of C57BLKS/J (BKS) origin. NOR IDDM resistance was previously found to be largely controlled by the Idd13 locus within an ∼24 cM segment on Chromosome 2 encompassing BKS-derived alleles for H3a, B2m, Il1, and Pcna. NOD stocks carrying subcongenic intervals of NOR Chromosome 2 were utilized to more finely map and determine possible functions of Idd13. NOR- derived H3a-Il1 (∼6.0 cM) and Il1-Pcna (∼1.2 cM) intervals both contribute components of IDDM resistance. Hence, the Idd13 locus is more complex than originally thought, since it consists of at least two genes. B2m variants within the H3a-Il1 interval may represent one of these. Monoclonal Ab binding demonstrated that dimerizing with the β2ma (NOD type) vs β2mb isoform (NOR type) alters the structural conformation, but not total expression levels of H2g7 class I molecules (e.g. Kd, Db). β2m-induced alterations in H2g7 class I conformation may partially explain findings from bone marrow chimera analyses that Idd13 modulates IDDM development at the level of non-hematopoietically derived cell types controlling selection of diabetogenic T cells and/or pancreatic β cells targeted by these effectors. Since trans-interactions between relatively common and functionally normal allelic variants may contribute to IDDM in NOD mice, the search for Idd genes in humans should not be limited to functionally defective variants.

Chromosome diversity in neotropical fishes: NOR studies

Neotropical fishes present a high chromosome diversity showing a wide diploid number variation range, including different levels of ploidies, sex chromosomes, chromosome supernumeraries, and several cases of polymorphisms, related particularly to heterochromatin and NOR sites. Two main general trends of chromosome diversification can be observed among neotropical fishes. First, several fish groups show a chromosome evolution relatively divergent from the point of view of the karyotypic macrostructure. Sister species show conspicuous differences in karyotype structure and most often also in the number of chromosomes. On the other hand, there are fish groups in which chromosome evolution has been shown to be less divergent, and in this case whole families or even groups of families may share a common karyotype structure and equal number of chromosomes. Several fish groups appear conservative also with respect to the NOR bearing chromosomes. In this case, NOR chromosome location is invariable among species. In contrast, several other groups present wide NOR variability. Sister species may show quite diverse chromosomes bearing nucleolar organizing regions. The NOR and heterochromatin relationship is also very diverse among fishes and this may indicate organizational differences involving these chromosome segments. Thus, neotropical fish fauna presents great chromosome variability, verifiable also by NOR studies.