Twenty years of physical mapping of major ribosomal RNA genes across the teleosts: A review of research.

Molecular cytogenetic data on the number and position of 45S ribosomal RNA genes (rDNA; located in nucleolus organizing regions, NORs) detected by FISH in 330 species of 77 families and 22 orders of bony fishes (Teleostei) and, additionally, 11 species of basal ray-finned fishes are compiled and analyzed. The portion of species with single rDNA sites in the sample amounts to 72%. The percentage of species with multiple NORs decreases with increasing numbers of rDNA loci per genome, i.e. scarcely 3% of species carry 4 or more rDNA-bearing chromosome pairs. 43% of all rDNA sites analyzed occur terminally on the short arms of chromosomes or constitute them. In general, terminal rDNA sites account for 87% of all examined cases. Interspecific variation in the location of single rDNA sites among related taxa, polymorphisms of multiple NORs in some groups of teleosts and analytical outcomes on the subject are reviewed.

Comparative cytogenetic study of two sister species of Iberian ground voles, Microtus (Terricola) duodecimcostatus and M. (T.) lusitanicus (rodentia, cricetidae).

The two Iberian species of pine voles, Microtus (Terricola) duodecimcostatus and M. (T.) lusitanicus of the subfamily Arvicolinae (Cricetidae, Rodentia), were compared after G- and C-banding and chromosomal mapping of ribosomal RNA genes (rDNA), telomeric repeats, and satellite DNA Msat-160. Notwithstanding their close relationship (one sister group in phylogenetic analyses) and sharing of the diploid and fundamental chromosome numbers, the 2 species show notable differences in the sex chromosome morphology, the number and distribution of rDNA sites, constitutive heterochromatin and satDNA patterns. The only telomeric repeats showed normal, all-telomeric, distribution in karyotypes of both species. The data are discussed with regard to interspecific and intrageneric variation of the analyzed characters and the chromosomal evolution in the genus Microtus.

Comparative cytogenetics of moles (Eulipotyphla, Talpidae): chromosomal differences in Talpa romana and T. europaea.

The genus Talpa is the most specious and widespread one in the family Talpidae. The existing karyological records are predominantly basic morphological descriptions. To further investigate the case in point, we performed a comparative cytogenetic study in the genus by comparing G- and C-chromosome banding and NOR patterns of the two European species, T. romana and T. europaea, along with available data regarding several other mole species. Chromosomal hybridization patterns for telomeric repeats and major and 5S ribosomal RNA genes were obtained in T. romana and T. europaea for the first time. The comparison of these patterns revealed differences in distribution of interstitial telomeric repeats and 5S ribosomal RNA genes in the two species with apparently identical karyotypes but different evolutionary histories.

Comparative molecular cytogenetic analysis of two congeneric species, Mugil curema and M. liza (Pisces, Mugiliformes), characterized by significant karyotype diversity.

Two congeneric mullet species, Mugil liza and M. curema, respectively with an all-uniarmed and an all-biarmed karyotype, were cytogenetically studied by base-specific fluorochrome staining and FISH-mapping of 45S and 5S ribosomal RNA genes (rDNA) and the (TTAGGG)(n) telomeric repeats. Whereas 45S rDNA sites might be homeologus in the two species, 5S rDNA sites are not, as they are localized on chromosome arms of different size. In both species, the (TTAGGG)(n) telomeric probe hybridized to natural telomeres and was found scattered along the NORs. In metacentric chromosomes of M. curema, no pericentromeric signals of the telomeric probe were detected. Data are discussed in relation to the karyotype evolution in Mugilidae and to the mechanisms and the evolutionary implications of Robertsonian rearrangements in M. curema.

FISH-mapping of 18S ribosomal RNA genes and telomeric sequences in the Japanese bitterlings Rhodeus ocellatus kurumeus and Tanakia limbata (Pisces, Cyprinidae) reveals significant cytogenetic differences in morphologically similar karyotypes.

The Japanese rose bitterling, Rhodeus ocellatus kurumeus, and the oily bitterling, Tanakia limbata, were cytogenetically studied by silver (Ag)- and chromomycin A3 (CMA3)-staining, by C-banding and by mapping of the 18S ribosomal genes and of the (TTAGGG)n telomeric sequence. These two representative species of related genera of the subfamily Acheilognathinae show very similar chromosome complements. Nevertheless, significant differences in the chromosomal distribution of nucleolus organizer regions (NORs) and interstitial telomeric sequences were observed. Whereas R. ocellatus kurumeus shows a single NOR-bearing chromosome pair, T. limbata is characterized by a higher number of variable NORs. Multiple telomeric sequence sites were found at the pericentromeric regions of several chromosomes in the rose bitterling. No telomeric sequence sites were detected near centromeres, but they were found to be scattered along the NORs in the oily bitterling. Two karyoevolutive trends might have been identified in the subfamily.

Classical and molecular cytogenetics of the zebrafish, Danio rerio (Cyprinidae, Cypriniformes): an overview.

The zebrafish, Danio rerio, has recently become the model system for the genetic analysis of vertebrate development. This paper reviews the advances in zebrafish cytogenetics, obtained through classical and molecular techniques, which will lead to the assignment of specific linkage groups to specific chromosome pairs in the zebrafish genome project. Several chromosome pairs of the 50-chromosome karyotype of D. rerio were differentially stained by classical staining techniques and additional information has been obtained by molecular cytogenetics. Indeed, the analysis of constitutive heterochromatin by C-banding and base-specific fluorochrome staining had suggested a differential composition of peri- and paracentromeric constitutive heterochromatin. The chromosome mapping of distinct AT- and GC-rich zebrafish satellite DNAs by means of PRINS (Primed in situ) and multicolor FISH (Fluorescence in situ Hybridization) has confirmed this hypothesis, which therefore provided the chromosome localization of 10% of the zebrafish genome. The analysis of nucleolus organizer regions (NORs) by silver staining and by FISH with 18S rDNA has also revealed the existence of variable and inactive NORs, in addition to those on the terminal regions of the long arms of the three NOR-bearing chromosome pairs. Other multicopy genes, such as minor ribosomal genes, or multicopy repeats, such as telomere specific sequences, have now been mapped on zebrafish chromosomes. The latest advancement in zebrafish molecular cytogenetics is the chromosome mapping of single locus genes. Single-copy genes from each of the 25 genetic linkage groups are now being mapped on zebrafish chromosomes by using PAC clones.

Chromosomal localization of zebrafish AluI repeats by primed in situ (PRINS) labeling.

Two zebrafish AluI repeats were localized in metaphase chromosomes by means of the primed in situ (PRINS) labeling technique, using oligonucleotide primers based on published sequences. An AT-rich, tandemly repeated, long AluI restriction fragment (RFAL1) labeled the (peri)centromeric regions of all chromosomes. The GC-rich short fragment (RFAS) was found to be localized in the paracentromeric regions of 17 chromosome pairs, which were mostly subtelocentric. The RFAS labeling pattern generally fits the previously described chromomycin A3 (CMA3) staining pattern. The differential composition of heterochromatin in zebrafish chromosomes is discussed.

Cytogenetic analysis of Liza ramada (Pisces, Perciformes) by different staining techniques and fluorescent in situ hybridization.

A cytogenetic investigation was carried out on specimens of Liza ramada, a mugilid species common in the Mediterranean sea. The analysis of chromosomes was performed through Ag-staining, C-banding, chromomycin A3 and DAPI staining, and fluorescent in situ hybridization with ribosomal genes. The results obtained are discussed with respect to cytotaxonomic implications and to the features of NORs.

CMA3-banding pattern and fluorescence in situ hybridization with 18S rRNA genes in zebrafish chromosomes.

This study provides new data on zebrafish chromosomes, obtained from the chromomycin A3-banding pattern and mapping of 18S rRNA genes by fluorescence in situ hybridization (FISH). C-banding and Agstaining were also performed to analyse whether variation in heterochromatin and Ag-nucleolus organizer regions (NORs) exists among various commercially purchased strains. The results provide information on heterochromatin composition and on the existence of interindividual NOR polymorphism and contribute to the construction of an idiogram suitable for gene mapping.

Cytogenetic analysis of a self-fertilizing fish, Rivulus marmoratus: remarkable chromosomal constancy over a vast geographic range.

The aplocheiloid killifish Rivulus marmoratus is the only known self-fertilizing hermaphroditic vertebrate. Most natural populations consist almost entirely of hermaphrodites and comprise arrays of homozygous clones. However, in almost all populations thus far studied, clonal variation, as detected with molecular techniques, is very high. A karyological survey was carried out on specimens from Brazil, the Bahamas, Belize, and Florida (4 locales) by C-banding, silver staining, and fluorescent staining. The chromosome complement of R. marmoratus is surprisingly uniform over its vast geographic range, in terms of both chromosome number and morphology, heterochromatin distribution and composition, and nucleolar organizer region (NOR) distribution. The short arms of chromosome pair 15, where NORs are located, showed the only variation detected in this study: those of pattern A were consistently shorter than those of pattern B; moreover, the latter show positive heteropycnosis with Giemsa staining. The present data demonstrate that chromosomal variation is not a significant part of the clonal divergence in this species, even though its breeding system, by forming homozygotes for new rearrangements almost immediately, should make that variation easier to detect. The high chromosomal homogeneity is discussed in the light of the peculiar natural history of the species.