[The peculiarities of the chromosome organization in meiosis].

Meiotic and mitotic chromosomes differ in a number of features. 1. At the early prophase I of meiosis, chromosomes acquire proteinaceous axial elements (AEs) which were absent at mitosis. In addition to somatic cohesins, AEs contain meiosis-specific cohesins REC8, SMC1beta, STAG3. 2. At the middle prophase I, proteinaceous lateral elements (LEs) of synaptonemal complexes (SC) are shaped on a basis of AEs. Proteins of LEs are not conserved, but in Saccharomyces cerevisiae and Arabidopsis thaliana they contain functional domains with conserved secondary structure. Proteins or functional domains similar to SC proteins have been found in green and brown algae, some of lower fungi and in Coelenterata amongs about 679 hundreds of proteins of primitive eukaryotes studied with bioinformatic methods. 3. During the pachytene and diplotene stages of meiosis, chromosomes of spermatocytes and mother pollen cells acquire the structure resembling in miniature the structure of amphibian and avian lamp brush chromosomes. Lateral chromatin loops of 90, 160 and more than 480 Kb in size are observed in human spermatocytes during the diplotene stage. Taken together, these findings support the idea of considerable conservation of the scheme of molecular and ultrastructural organization of meiotic chromosomes in a variety of eukaryotic organisms.

[Morphological manifestation of a unique DNA segment in human meiotic prophase I].

The commercial sample of human DNA fragment from the choromosome 17 was used as the probe for FISH to study of the mode of its attachment to the lateral elements of synaptonemal complex (SC) in human spermatocytes. It was a 160 kb probe from the band 17p1.2, containing RAI1 gene with D17S620 marker (the probe for deletion causing Smith-Magenis syndrome). The probe made lateral chromatin protrusions, contacting with SC stained with anty-SYCP3. Different morphological configuration of lateral chromatin protrusions where observed. They depended on substages of meiotic prophase I. At zygotene, FISH probe form two sticks, c. a. 6 micro long, which was perpendicular to SC longitudinal axe, one stick at each SC side. At early pachytene, each stick transforms into a globule, one globule at each SC side again. At late pachytene each globule transformed into two crumbly globules containing short threads and clumps. At diplotene, globules finally transformed into thin DNA (chromatin) loops up to 10 micro long from the base to top with periodical thickenings (beads) along their length. As the result of this dynamics of transformation, two chromatin loops with beads were found on each side of SC of the chromosome 17. These loops most probably were the loops of sister chromatides, the full set of chromatide loops at the particular SC (bivalent) site being four in number, i. e. representing of two pair of chromatides. This study is the first one in which lateral chromatin loops in human mail meiotic prophase I are visualized as true open loop instead of that usually postulated "loops" after observation of condensed road-like or brush-like chromatin protrusion attached to the lateral elements of synaptonemal complexes. Open configuration of the loops, presumably, depends on activation of transcription during late pachytene-early diplotene. They resemble lateral loops of mini lampbrush chromosomes.

[DNA repeated sequences may be involved in the synaptonemal complexes formation].

Synatonemal complexes (SCs) are the intranuclear structures which facilitate reversible lateral synapsis of the homologous chromosomes in the course of meiosis. It is still unclear which DNA nucleotide sequences are responsible for the chromatin attachment to the SC lateral elements. Considering the features of the dispersed repeated sequences (RS) it is worth to assume their participation in the structure functional organization of the meiotic chromosome. Using numerical analysis we have investigated the relationship between RS and the distribution of events of the meiotic recombination in mouse chromosome 1. Using in situ hybridization on spread mouse spermatocytes, we have demonstrated the arrangement of different types of RS relative to SCs. Hybridization signals of B1(Alu), B2, and minisatellite probes were localizating predominantly in the SCs regions. Our results allow us to suggest the model of the meiotic chromosome organization with the RS as the sequences, participating in the attachment of chromatin loops and SCs.

[How do chromosomes attach to synaptonemal complexes?].

Fluorochrome-labeled oligonucleotides (n = 44) corresponding to mouse genome repetitive sequences were hybridized in situ with pachytene nuclei of mouse spermatocytes. Signals of the repetitive sequences MaLR, MER, and (GT)22 were found to be dispersed through chromatin, and signals of BI 1 repeats and minisatellites were mostly attached to synaptonemal complexes immunostained with anti-SYCP3 antibodies. These results suggest that B 1 repeats and minisatellites are candidates for sequences anchoring chromatin to synaptonemal complexes.

[Developing and testing a method of in silico identification and characterization of meiotic DNA].

A method of in silico search for specific repetitive DNA sequences related to the synaptonemal complex (meiDNA) in mammalian genomes was developed. A study of the distribution of these repeats over chromosomes revealed their scarcity on the Y chromosome and a decrease in recombination frequency in regions enriched in meiDNA. The results are discussed in context of the model of the looplike meiotic chromosome organization during the formation of the synaptonemal complex.

[In silico identification and characterization of meiotic DNA: AluJb possibly participates in the attachment of chromatin loops to synaptonemal complex].

Earlier, using bioinformatic methods, we reported the identification of repeated DNA sequences (RS), presumably responsible for the attachment of chromatin loops to the lateral elements of synaptonemal complex in meiotic chromosomes. In the present study, consensus sequences for this class of RS were identified. It was demonstrated that at least part of these sequences belonged to the AluJb subfamily of Alu sequences. The Alu copies distribution along the major human histocompatibility complex (MHC) and their spatial separation from the sites of meiotic recombination was examined. It was demonstrated that simple sequences, like (GC/CA)n, were flanking meiotic recombination sites. A model of the RS organization in meiotic chromosome, most efficiently linking experimental data on the meiotic recombination in MHC and the in silico data on the RS localization (the coefficient of multiple correlation, r = 0.92) is suggested.

[On the question of using alleles of gliadin-coding loci as possible markers of adaptive ability in cultivars of spring wheat (Triticum aestivum L.) during seed germination]. / K voprosu ispol'zovaniia allelei gliadinkodiruiushchikh lokusov v kachestve vozmozhnykh markerov adaptivnosti u sortov iarovoi miagkoi pshenitsy (Triticum aestivum L.) v protsesse prostrania zerna.

Prolamine proteolysis is assumed to be among numerous adaptability factors in cereals. The patterns of gliadin proteolysis have been studied in 16 cultivars of spring wheat via analysis of electrophoretic spectra. Four proteolytic patterns have been identified. It is hypothesized that the cultivars characterized by early and rapid proteolysis (the first and third types) are the most adaptable. The gliadin genetic formulas of chromosomes of the first homeologous group have been determined. The alleles of gliadin loci (Gli-Alf, Gli-Ble, Gli-D1a, and Gli-D1b) have been found that can be used as markers of adaptability in spring wheat cultivars.

[Comparative genomics and proteomics of Drosophila, Brenner's nematode, and Arabidopsis: identification of functionally similar genes and proteins of meiotic chromosome synapsis]. / Sravnitel'naia genomika i proteomika Drozofily, nematody Brennera i Arabidopsisa. Identifikatsiia funktsional'no skhodnykh genov i belkov sinapsisa meioticheskikh khromosom.

The published principles of computer analysis of genomes and protein sets in taxonomically distant eukaryotes are expounded. The authors developed a search strategy to identify in genomes of such organisms genes and proteins nonhomologous in primary structure but having similar functions in cells dividing by meiosis. This strategy based on the combined principles of genomics, proteomics, and morphometric analysis of subcellular structures was applied to a computer search for genes encoding the proteins of synaptonemal complexes in genomes of Drosophila melanogaster, the nematode Caenorhabditis elegans, and the plant Arabidopsis thaliana. These proteins proved to be functionally similar to their counterparts in yeast Saccharomyces cerevisiae (protein Zip1p) and mammals (protein SCP1).

[CG17604 gene from Drosophila melanogaster--possible functional homolog of the yeast ZIP1 and SCP1 (SYCP1) mammalian genes, coding for synaptonemal complex proteins]. / Gen CG17604 Drosophila melanogaster--vozmozhnyi funktsional'nyi gomolog genov ZIP1 drozhzhei i SCP1 (SYCP1) mlekopitaiushchikh, kodiruiushchikh belki sinaptonemnogo kompleksa.

From data on the molecular organization of transverse filament proteins of the synaptonemal complex (SC)--Zip1 in yeast and SCP1 in mammals--and on the width of the central SC space in these organisms and in Drosophila, the putative molecular structure and size of a transverse filament protein of the SC in Drosophila has been inferred. Using genetic and molecular databases and software from the Internet, we carried out in silico screening for a candidate gene for the Drosophila transverse filament protein. The search in the 250-bp region overlapping the locus of this gene (sections 88E-89B) and containing 78 predicted genes has revealed only one gene, CG17604, whose protein meets all requirements for the transverse filament protein of the SC. It was suggested that gene CG17604 is gene c(3)G. In this case, gene c(3)G must be localized in section 89A7-8 of the cytological map of Drosophila melanogaster.

[Damaging effect of antibiotics on the structure of synaptonemal complexes of meiotic chromosome of mice]. / Povrezhdaiushchee deistvie antibiotikov na strukturu sinaptonemnykh kompleksov meioticheskikh khromosom myshi.

The structure of synaptonemal complexes (SCs) of chromosomes of mouse primary spermatocytes were studied using electron microscopy on days 1, 10, and 36 after the completion of per os administration of drugs belonging to three groups of antibiotics: tetracyclins, macrolides, and fluoroquinolones. The antibiotics were administered to mice during ten days. At the substages of early and middle pachytene, heteromorphic SC bivalents and fragments of chromosome-core elements were detected in spermatocytes at all times studied after the administration of the antibiotics of three groups. As cells passed through the period from early to middle pachytene, the number of cells containing heteromorphic SC bivalents and the fragments of chromosome cores gradually decreased, which could be an indication of selection of cells with chromosomal aberrations. A high level of associations between the X chromosome and autosome bivalents (including heteromorphic ones) also favors this suggestion. A gradual decrease in the number of chromosomal aberrations was detected, as time elapsed from the completion of antibiotics administration. The study of sperm obtained from epididymises of males did not reveal significant differences in both morphology and motility of sperm between males of the control and experimental groups.

[Use of artificial neuronal nets in automation of analysis and genetic identification of electrophoretic spectra of gliadin from durum wheat]. / Ispol'zovanie iskusstvennykh neironnykh setei pri avtomatizatsii analiza i geneticheskoi rasshifrovke élektroforeticheskikh spektrov gliadina tverdoi pshenitsy.

Each wheat cultivar has a characteristic spectrum of gliadins. This makes it possible to use blocks of the components of reserve proteins as genetic markers when estimating seed quality. However, identification of the blocks that constitute the electrophoretic spectrum is a complicated task. For this purpose artificial neural network (ANN) technology is proposed. Based on experimental data, a teaching database and testing databases have been created. ANN was shown to be highly efficient (efficiency up to 100%) expert system for deciphering the electrophoretic spectra of gliadins of durum wheat cultivars.

[The dynamics of the formation of spontaneous autoantibodies to the synaptonemal complex in male mice]. / Issledovanie dinamiki formirovaniia spontannykh autoantitel k sinaptonemnomu kompleksu u samtsov myshi.

Time-related changes in formation of spontaneous autoantibodies against the synaptonemal complex were studied by indirect immunofluorescence in male mice. Appearance of the spontaneous autoantibodies against the synaptonemal complex coincided with that of the cells containing synaptonemal complexes. The mouse synaptonemal complexes were binding spontaneous autoantibodies of the rabbit and human sera. The synaptonemal complexes of the equine spermatocytes were binding spontaneous autoantibodies of the mouse serum. There was no fluorescence of synaptonemal complexes on preparations of spread rye meiocytes treated with the mouse serum. Antigenic similarity was shown for the synaptonemal complex components in representatives of the different mammalian orders: rodents, odd-toed ungulates, and primates.

[Features of the primary structure of DNA from the synaptonemal complex of the golden hamster]. / Osobennosti pervichnoi struktury DNK iz sinaptonemnogo kompleksa zolotistogo khomiachka.

DNA of golden hamster synaptonemal complex (SC) was cloned and partially sequenced. Computer analysis was performed to compare the SC DNA sequences for different families of eukaryotic genomic DNA. SC DNA is homologous to LINE family and in contrast to genomic DNA is enriched with alternative (GT/CA)n sequences and palindromes. By its ability to form potential hairpin loops, SC DNA is similar to nuclear matrix-associated regions of DNA (MARs) and differs considerably from DNA of microsatellite loci. It is noteworthy that SC DNA, which is supposed to be involved in meiotic recombination, and DNA of immunoglobulin genes, involved in somatic recombination, are both enriched with palindromes. The discovered peculiarities of the structure of hamster SC DNA allow us to join it with rat SC DNA into a special family of rodent genomic DNA sequences.

[Autoantibody formation against the antigens of the synaptonemal complex in the syngeneic immunization of male Mus musculus]. / Obrazovanie autoantitel protiv antigenov sinaptonemnogo kompleksa pri singennoi immunizatsii samtsov myshei Mus musculus.

Male mice were immunized with the suspension of synaptonemal complexes (SC) isolated from mouse spermatocytes nuclei. The indirect immunofluorescent analysis showed the active binding of sera obtained from immunized mice to SC of mouse spermatocyte spreads. At early and mid-pachytene, SC can be clearly identified in 19 autosome bivalents and in sex chromosome bivalent. According to the electron microscopic analysis, all structural elements of SC bind antibodies. Metaphase chromosomes were not stained with the immune sera. Specificity of interaction between SC components and antibodies was confirmed in a series of control experiments. Analysis of sera obtained from mice after their syngeneic immunization with isolated SC fraction suggested that certain mouse SC components induce the formation of autoantibodies. This, in turn, suggests that these SC components are meiosis-specific.

[A preparative method for isolating the synaptonemal complexes from mammalian spermatocytes]. / Preparativnyi metod vydeleniia sinaptonemnykh kompleksov iz spermatotsitov mlekopitaiushchikh.

A method of isolation of synaptonemal complexes (SC) from mouse, rat and Syrian hamster spermatocytes is described. A fraction of pachytene spermatocyte nuclei was obtained by centrifugation of the testis homogenate in stepwise sucrose gradient and then lysed. The resulting chromatine was hydrolysed with DNAse II, and a fraction of isolated SCs was obtained by ultracentrifugation of the hydrolysate. The method can be applied for obtaining the SC fraction from spermatocytes sufficient for cytological, biochemical and molecular biology studies.