[Comparative cytogenetics of main Laurasiatheria taxa].

This short communication is a review of key trends in the karyotypic evolution of mammalian taxa Laurasiatheria, inferred from comparative chromosome painting.

Identification of all pachytene bivalents in the common shrew using DAPI-staining of synaptonemal complex spreads.

A major problem in studies of synaptonemal complexes (SC) is the difficulty in distinguishing individual chromosomes. This problem can be solved combining SC immunostaining with FISH of chromosome-specific sequences. However, this procedure is expensive, time-consuming and applicable only to a very limited number of species. In this paper we show how a combination of SC immunostaining and DAPI staining can allow identification of all chromosome arms in surface-spreads of the SC of the common shrew (Sorex araneus L.). Enhancement of brightness and contrast of the images with photo editing software allowed us to reveal clear DAPI-positive and negative bands with relative sizes and positions similar to DAPI landmarks on mitotic metaphase chromosomes. Using FISH with DNA probes prepared from chromosome arms m and n we demonstrated correct recognition of the chromosomes mp and hn on the basis of their DAPI pattern. We show that the approach we describe here may be applied to other species and can provide an important tool for identification of individual bivalents in pachytene surface-spreads.

[Interstitial telomere repeats as markers of evolutionary changes in the mammalian karyotype: human chromosome 2].

Telomer repeats represented by hexamer (TTAGGG)n at chromosome termini are required for correct function and chromosome stability. At the same time, interstitial telomer sequence (ITS) located far from the chromosome ends are known for several mammalian genomes, including the human genome. It is assumed that these repeats mark the points of fusion or other chromosome reconstructions of ancestors. Exact localization of all interstitial telomer sequences in the genome could greatly improve our understanding of the mechanism of karyotype evolution and species origin. We have developed a software for a search of interstitial telomer sequences in complete sequences of mammalian genomes. We have demonstrated the evolutionary significance of repeats by an example of human chromosome 2. The results and supplementary materials are available at the site of the Institute of Cytology and Genetics: http://www.bionet.nsc.ru/labs/theorylabmain/orlov/telomere/.

Comparative map between the domestic pig and dog.

Cross-species chromosome painting with probes derived from flow-sorted dog and human chromosomes was used to construct a high-resolution comparative map for the pig. In total 98 conserved autosomal segments between pig and dog were detected by probes specific for the 38 autosomes and X Chromosome of the dog. Further integration of our results with the published human--dog and cat--dog comparative maps, and with data from comparative gene mapping, increases the resolution of the current pig--human comparative map. It allows for the conserved syntenies detected in the pig, human, and cat to be aligned against the putative ancestral karyotype of eutherian mammals and for the history of karyotype evolution of the pig lineage to be reconstructed. Fifteen fusions, 17 fissions, and 23 inversions are required to convert the ancestral mammalian karyotype into the extant karyotype of the pig.

Comparative molecular cytogenetic studies in the order Carnivora: mapping chromosomal rearrangements onto the phylogenetic tree.

We have made a set of chromosome-specific painting probes for the American mink by degenerate oligonucleotide primed-PCR (DOP-PCR) amplification of flow-sorted chromosomes. The painting probes were used to delimit homologous chromosomal segments among human, red fox, dog, cat and eight species of the family Mustelidae, including the European mink, steppe and forest polecats, least weasel, mountain weasel, Japanese sable, striped polecat, and badger. Based on the results of chromosome painting and G-banding, comparative maps between these species have been established. The integrated map demonstrates a high level of karyotype conservation among mustelid species. Comparative analysis of the conserved chromosomal segments among mustelids and outgroup species revealed 18 putative ancestral autosomal segments that probably represent the ancestral chromosomes, or chromosome arms, in the karyotype of the most recent ancestor of the family Mustelidae. The proposed 2n = 38 ancestral Mustelidae karyotype appears to have been retained in some modern mustelids, e.g., Martes, Lutra, Ictonyx, and Vormela. The derivation of the mustelid karyotypes from the putative ancestral state resulted from centric fusions, fissions, the addition of heterochromatic arms, and occasional pericentric inversions. Our results confirm many of the evolutionary conclusions suggested by other data and strengthen the topology of the carnivore phylogenetic tree through the inclusion of genome-wide chromosome rearrangements.

[A new approach to the analysis of complex chromosomal rearrangements in cell hybrids]. / Novye vozmozhnosti analiza slozhnykh khromosomnykh perestroek v kletochnykh gibridakh.

The chromosomal complements of somatic cell pig-mink hybrids was determined by a new approach. This approach includes microdissection of metaphase chromosomes, generation of chromosome and region-specific DNA libraries, and fluorescence in situ hybridization of these libraries with pig lymphocyte chromosomes. The studied hybrid cells were shown to contain two small acrocentric chromosomes and a microchromosome of porcine origin. Identification of these chromosomes by differential GTG-staining was impossible. Chromosome isolation by a micromanipulation technique followed by DNA amplification in TOPO-DOP polymerase chain reaction provided chromosome-specific DNA libraries of the rearranged chromosomes. Based on these libraries, the labeled DNA probes were prepared and hybridized to pig chromosomes. This allowed us to determine the origin of the material contributing to the hybrid cell chromosomes. One of these chromosomes contained five pig chromosomal regions: 15cen-q2; 6q21-q23; 13q21; 13q22; 7q25-qter, while the other contained the following pig chromosomal regions: 4p12-p13; 16q12-q14; 12pter-p15. The microchromosome contained the Xp11-Xq11 region. The minimal size of the revealed chromosomal regions was about 3 to 4 x 10(6) bp. Segregation analysis of the thymidine kinase gene 1 (TK1), which was earlier localized to the pig 12p region, and the hybrid cell pig chromosomes in the hybrid subclones suggested that TK1 gene can be assigned to 12p15-pter. The results obtained demonstrate the efficiency of the applied approach in its detailed and reliable description of complex chromosomal rearrangements in hybrid clones, when differential chromosome staining failed to identify these chromosomes.

[Feasibility of analysis of macroevolutionary rearrangements of mammalian chromosomes]. / Vozmozhnosti analiza makroévoliutsionnykh preobrazovanii khromosom mlekopitaiushchikh.

In the Institute of Cytology and Genetics, Siberian Division, Russian Academy of Sciences, detailed studies on chromosome sets of humans and domestic animals were initiated and supported by D.I. Belyaev and started by S.I. Radzhabli. They believed that analysis of differentially stained chromosomes and mapping of the genomes of main commercial species provide for a better understanding of the processes that occurred during their evolution and domestication. Several new approaches to studying macroevolutionary karyotypic rearrangements associated with divergence of remote taxa, such as primates and paridigitate ungulates, are discussed.

Localization of the genes for major ribosomal RNA on chromosomes of the house musk shrew, Suncus murinus, at meiotic and mitotic cells by fluorescence in situ hybridization and silver staining.

The genes for major ribosomal RNA were localized on chromosomes 5pter-p15, 9q64-qter, and 13q38-qter of the house musk shrew, Suncus murinus (Insectivora, Soricidae) by silver staining of mitotic metaphase and meiotic pachytene spreads and fluorescence in situ hybridization using the human 28S-RNA genes as a probe to mitotic metaphase spreads. The data presented indicate a correlation between sites of in situ hybridization and silver staining. The finding of nuclear materials in mitosis was in a good agreement with observation in meiosis: same chromosomes carried active NORs in both meiotic and mitotic cells.

Localization of rat K51 keratin-like locus (Krt10l) to human and animal chromosomes by in situ hybridization.

The rat K51 locus (gene symbol Krt10l) was mapped using isotopic in situ hybridization to rat chromosome 3, human chromosome 9, pig chromosome 6, cattle chromosome 18, and mink chromosome 1.

Localization of ESD and A2M genes to sheep chromosome 3 by in situ hybridization.

The genes encoding esterase D (ESD) and alpha 2-macroglobulin (A2M) were mapped using 3H-labeled cDNAs to sheep chromosome 3p26-->p24 and 3q26-->q35 respectively.

[Homology of G-banded mammalian chromosomes]. / Gomologiia G-okrashennykh khromosom mlekopitaiushchikh.

Linkage relationships of homologous loci and high resolution G-banding patterns of man, mouse, rat, chinese hamster, rabbit, cat, mink, pig, ox and sheep were used for identification of 11 evolutionary conservative autosomal regions. The distributions and inversions of these regions in the ancestor genomes of some phylums have been shown. For example, the regions homologous to human Ip region were detected in cat, mink and rabbit genomes. In the genomes of rodents studied the intraregion inversion was detected. In the ox and sheep genomes the distal end deletions were detected within these regions. In the pig genome these regions were represented solely by disruptions. We supposed that the rapid "catastrophic" chromosomal evolution took place during short time periods of some orders and families separation.