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O L Kolomiets

Publications and source records attributed to O L Kolomiets.

At least 19 recordsLinked to original sources

[Abnormal meiosis in bisporic mutants of white button mushroom Agaricus bisporus (Lange) Imbach].

A formerly developed method of obtaining spread preparations of mushroom basidial nuclei was applied to study of meiotic prophase I in bisporic white button mushroom (Agaricus bisporus) strains. Meiotic recombination and assemblage of axial structures (axial elements and synaptonemal complexes) of chromosomes in meiotic prophase I are interrelated. It is known that the frequency of meiotic recombination is reduced in the bisporic A. bisporus variety. We showed that formation of axial structures of meiotic chromosomes in bisporic strains of this mushroom was disrupted. The phenotypes of disruptions in spread prophase nuclei are diverse. In leptotene and early zygotene, many nuclei contain abnormal, often short, and, as a rule, few chromosomal axial elements. The abnormalities in the formation of synaptonemal complexes at the zygotene-diplotene stage are of the same kind and even more pronounced. We discovered an important feature of meiosis in A. bisporus associated with fruit-body morphogenesis. Meiosis starting in basidia (meiocytes) of young closed fruit bodies is accompanied by disruption of chromatin condensation in prophase I and, probably, is arrested. After indusium breakage, the course of meiosis normalizes. Preparations with clearly observable chromosomal axial structures can be obtained only at this stage of fruit-body development.

Agaricus↗

Meiotic mutations in rye Secale cereale L.

Spontaneous meiotic mutations of winter rye Secale cereale L. (2n = 14) were revealed in inbred F2 progenies, which were obtained by self-pollination of F1 hybrids resulting from crosses of individual plants of cultivar Vyatka or weedy rye with plants of self-fertile inbred lines. The mutations cause partial or complete sterility, and are maintained in heterozygote condition. Six types of mutations were distinguished as the result of cytological analysis of meiosis and genetic analysis. (1) Plants with nonallelic asynaptic mutations sy1 and sy9 lacked bivalents in 96.8 and 67.0% metaphase I cells, respectively, formed only axial elements but not the mature synaptonemal complex (SC), and had defects in telomere clustering in early prophase I. (2) Weak asynaptic mutant sy3 showed incomplete synapsis at the start of SC degradation at diplotene and lower chiasma number; yet only 2% meiocytes lacked bivalents in MI. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19 caused nonhomologous synapsis; i.e., a varying number of univalents and occasional multivalents were observed in MI, which was preceded by switches of pairing partners and fold-back synapsis at mid-prophase I. (4) Mutation mei6 led to the formation of protrusions and minor branched structures of the SC lateral elements. (5) Allelic mutations mei8 and mei8-10 caused irregular chromatin condensation along the chromosome length in prophase I, which was accompanied by chromosome sticking and fragmentation in MI. (6) Allelic mutations mei5 and mei10 determined chromosome supercondensation, caused the disturbance of meiotic spindle assembly, arrested meiosis at various stages but did not affect formation of the pollen wall, thus arrested meiocytes got covered with the pollen wall. Analysis of double mutants revealed recessive epistatic interactions for some mutations; the epistatic group was sy9 > sy1 > sy3 > sy19. This reflects the sequence of meiotic events controlled by the corresponding genes. The expression of sy2 and sy19 proved to be modified by additional genes. Most meiotic mutations found in rye have analogs in other plants.

Chromosome Pairing↗

[Genetic collection of meiotic mutants of rye Secale cereale L].

Genetic collection of meiotic mutants of winter rye Secale cereale L. (2n = 14) was created. Mutations were detected in inbred F2 generations after self-fertilization of the F1 hybrids, obtained by individual crossing of rye plants (cultivar Vyatka) or weedy rye with plants from autofertile lines. The mutations cause partial or complete plant sterility and are maintained in collection in a heterozygous state. Genetic analysis accompanied by cytogenetic study of meiosis has revealed six mutation types. (1) Nonallelic asynaptic mutations sy1 and sy9 caused the formation of only axial chromosome elements in prophase and anaphase. The synaptonemal complexes (SCs) were absent, the formation of the chromosome "bouquet" was impaired, and all chromosomes were univalent in meiotic metaphase I in 96% (sy1) and 67% (sy2) of cells. (2) Weak asynaptic mutation sy3, which hindered complete termination of synapsis in prophase II. Subterminal asynaptic segments were always observed in the SC, and at least one pair of univalents was present in metaphase I, but the number of cells with univalents did not exceed 2%. (3) Mutations sy2, sy6, sy7, sy8, sy10, and sy19, which caused partially nonhomologous synapsis: change in pairing partners and fold-back chromosome synapsis in prophase I. In metaphase I, the number of univalents varied and multivalents were observed. (4) Mutation mei6, which causes the formation of ultrastructural protrusions on the lateral SC elements, gaps and branching of these elements. (5) Allelic mutations mei8 and mei10, which caused irregular chromatin condensation along chromosomes in prophase I, sticking and fragmentation of chromosomes in metaphase I. (6) Allelic mutations mei5 and mei10, which caused chromosome hypercondensation, defects of the division spindle formation, and random arrest of cells at different meiotic stages. However, these mutations did not affect the formation of microspore envelopes even around the cells, whose development was blocked at prophase I. Analysis of cytological pictures of meiosis in double rye mutants reveled epistatic interaction in the mutation series sy9 > sy1 > sy3 > sy19, which reflects the order of switching these genes in the course of meiosis. The expression of genes sy2 and sy19 was shown to be controlled by modifier genes. Most meiotic mutations found in rye have analogs in other plant species.

Chromatin Assembly and Disassembly↗

[The effect of mutation dominant spotting-Yurlovo (KitW-Y) on spermatogenesis, early embryogenesis, and fertility of C57BL/6JY mice].

The effect of mutation KitW-Y found in C57BL/6 mice on fertility, spermatogenesis, and early embryogenesis of mice have been studied. If heterozygotes KitW-Y/+ are crossed with wild-type mice, fertility decreases by 20%. Homozygotes Kitw-Y/KitW-Y and compounds KitW-Y/KitSsm are nonviable. The study of spermatogenesis in KitW-Y/+ mice has demonstrated a negative effect of this mutation on spermatocytes. Histological examination of the testes of mutant males has shown local empty spaces in seminal ducts. Electron microscopic examination of synaptonemal complexes have demonstrated desynapsis disturbance in some nuclei at the diplotene stage of meiotic prophase I. However, these disturbances do not cause a decrease in the number of fertilized oocytes/ova. The decrease in fertility is accounted for disturbances of early embryogenesis. In vivo and in vitro analyses of early embryogenesis have demonstrated that cleavage divisions are asynchronous in KitW-Y/+ heterozygous embryos. Some of these embryos die before implantation, and others cleave more rapidly than wildtype embryos, which give them selective advantage during the postimplantation period of embryogenesis. The pattern of KitW-Y expression during spermatogenesis and embryogenesis mimics potential human pathology, which makes these mutants an interesting and valuable object for genetics and developmental biology.

Animals↗

Destructive effect of DNA topoisomerase II inhibitor vepesid on mouse spermatogenesis.

Inhibition of DNA topoisomerase II with vepesid induced structural and functional reorganization of chromatin in meiotically dividing spermatocytes I, which later led to the block of their differentiation and long-lasting disorders in spermatogenesis. Vepesid induced decondensation of spermatocyte I chromatin, block of desynapsis, and elongation of lateral elements of spermatocyte autosome synaptonemal complexes during late pachytene and diplotene of meiosis. This confirms the involvement of type II DNA topoisomerase in chromatin condensation and homologous chromosome desynapsis at the stage of diplotene and the role of this enzyme in structural organization of the synaptonemal complex. Vepesid induced the formation of dichotomy and breaks of the pericentromer regions of subelements of lateral elements of the autosomal synaptonemal complexes; the number of cells with associations of axial elements of sex chromosomes with autosomal synaptonemal complexes increased, univalents of autosomes and sex chromosomes appeared. Mesna, a modifier of toxic effects of antitumor drugs, had no toxic effect on spermatogenic cells. Mesna reduced the lethal effect of vepesid during combined treatment, but did not ensure long-term protection of spermatogenesis.

Animals↗

[A method for preparation of synaptonemal complexes of meiotic chromosomes from basidial protoplasts of the common mushroom Agaricus bisporus (Lange) Imbach].

For the first time, preparations of synaptonemal complexes (SCs) were made from meiotic chromosomes of white button mushroom (Agaricus bisporus) basidia. It is the first experience of obtaining SC preparations of filamentous fungi from isolated meiosporangium protoplasts. Previously, only yeast SC preparations were obtained following this approach. The method includes four major stages: isolation of basidium protoplasts by treatment of basidia with lytic enzymes, spreading of protoplast nuclei on a filmy support by osmotic shock, staining the preparations with silver nitrate, and examination under light and electron microscopes. The structures of spread premeiotic nuclei, axial elements of chromosomes, SCs, chromatin, and nucleoli were studied at the leptotene-diplotene stage of meiotic prophase I.

Agaricus↗

Damaging effect of taxol on mouse spermatogenesis.

Taxol produced a specific effect on mouse spermatocytes I and on stem spermatogonia, which leads to overall degeneration of spermatocytes I and long lasting disorders of spermatogenesis. Breaks of the axial and lateral elements of the synaptonemal complex, aneuploidies in spermatocytes in early, middle, and late pachytene, and accumulation of cells with associations of sex chromosomes and autosomes were observed, which attested to blockade of spermatogenesis in late pachytene and diplotene of meiosis prophase I. Mesna, a chemoprotective agent, reduced total toxicity and lethal effects of taxol, but did not prevent destruction of the testes.

Animals↗

[Genetic control of chromosomal synapsis in Secale cereale L. rye meiosis: sy19 gene, causing heterologous synapsis].

Analysis of manifestation and inheritance of a new mutation inducing irregular synapsis in rye showed that abnormal phenotype is determined by a recessive allele of the sy19 gene. In the homozygotes for this mutation, even at the light microscopic level, abnormal formation of bivalents is already observed at pachytene-diakinesis. At metaphase I, the univalent frequency varies from 0 to 14; in a few cells, multivalent associations of chromosomes, which are not clearly oriented in the spindle, are detected. Electron microscopy of synaptonemal complexes revealed both homologous and heterologous synapsis in homozygotes for sy19, namely partial loss of the ability to stringent homology search. Analysis of joint inheritance of sy19 and asynaptic sy1 mutations showed that they are nonallelic, inherited independently, and interact by recessive epistasis. The phenotype of double sy1sy19 mutants indicates that the sy19 gene conditioning heterologous synapsis operates at meiosis later than the synaptic gene sy1. The epistatic group of mutations, sy9 > sy1 > sy19 and sy3, was determined.

Chromosomes↗

[Damaging effect of antibiotics on the structure of synaptonemal complexes of meiotic chromosome of mice].

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.

Animals↗

[Non-Mendelian inheritance of the Un(s) mutant gene in mice].

The phenotypic expression of the mutation Uns (gene Pax-1) were studied in the progeny of heterozygous mice. In all crosses, the sex ratio close to 1:1 was observed in the progeny. However, among sons and daughters the segregation for the mutant character depended on the variant of the cross variant. When the Uns/+ mice were intercrossed, the ratio of mutant to normal females was 3:1, while the corresponding ratio among males was 0.9:1. Studying preimplantation development in vitro and postimplantation development in vivo revealed no significant mortality in embryos obtained by crossing heterozygous mice. Ultrastructural analysis revealed no distortions in synapsis and desynapsis of axial elements of the autosomal chromosomes in Uns/+ mice. The segregation found seems to result either from different fertilizing ability of normal sperm and sperm carrying the Uns mutation or, which is more probable, from the genetic imprinting of the gene Uns.

Animals↗

[Localization of RecA-like proteins in preparations of spread nuclei of mouse spermatocytes at meiotic prophase I].

Earlier, polyclonal antibodies to Escherichia coli RecA protein were used to identify immunologically related proteins in meiotic spermatocytes of different eukaryotes. At least one such protein proved to be a component of the synaptonemal complex (SC) [1]. Subsequent experiments on localization of RecA-like antigens in SCs of spermatocytes were performed by indirect immunocytochemical methods and electron microscopy, which showed that RecA-like protein(s) at early leptotene are largely associated with chromatin. During SC formation (at leptotene and zygotene), they are found in both lateral elements and the central space of SC. In some cases, RecA-like proteins are associated with SC substructures that resemble recombination nodules. When spermatocytes enter late diplotene, Rec-A-like proteins cease to be detected in SC structures.

Animals↗

[Cytogenetic patterns in synapsis of meiotic chromosomes in animals and plants].

Chromosome synapsis in meiosis is performed through the formation of a synaptonemal complex (SC). The SC aggregates homologues until the diplotene stage, when the function of physical binding is carried out by a chiasmata. SC formation is controlled by specific genes. Regularities of SC formation in animals and plants are being studied at the Laboratory of Cytogenetics, Vavilov Institute of General Genetics, the Russian Academy of Sciences. It was shown that, in Drosophila melanogaster, the presence or absence of SC in meiosis is determined by balance between autosomes and sex chromosomes. i.e., SC is formed according to regularities in sex determination. The dose of the 8C-11A segment of X chromosome plays a key role in this process. Eight mutations in synaptic genes that affect SC formation were studied in rye Secale cereale. SC variants occurring in heteromorphic sex chromosomes were analyzed in the Ellobius genera and in B-chromosomes of Apodemus peninsulae. It was shown that, in addition to homologous synapsis, non-homologous synapsis of aberrant chromosomes occurs in rye and mouse heterozygotes at chromosome rearrangements. Twice as many heterozygous chromosome rearrangements were revealed, when the SC was studied at pachytene instead of diakinesis and metaphase I. Simple methods of SC investigations extended the capabilities of cytogenetic analysis of meiosis and provided new perspectives for studying the action of meiotic gen.

Animals↗

[Antibodies against Escherichia coli RecA protein reveal two nuclear proteins in bovine spermatocytes which interact with synaptonemal complex structures of meiotic chromosomes of various eukaryotic organisms].

Three RecA-like proteins were detected in bovine meiotic cells using antibodies against Escherichia coli RecA protein. After isolation and purification of these RecA-like proteins their molecular weights appeared to be equal to 37, 70 and 130 kD. The 37 kD protein accompanies all the stages of spermatogenesis up to the stage of mature spermatozoa. The 70 kD protein is detectable only in nuclei of cells at the stage of prophase I of meiotic division. These RecA-like proteins are involved in the formation of structural elements of the synaptonemal complex (SC) and are detected in the SC composition in meiotic cells not only of mammals but also of plants and insects, which suggests the evolutionary conservative character of these proteins.

Animals↗

[Synaptonemal complexes of chromosomes of male mice after prolonged administration of neoaquasept].

A long-term (during 4 months) daily peroral injections of neoaquasept (NA) (a drinking water disinfections agent) at a dose corresponding to the normal level of consumption increases the number of spermatocytes in which the sexual bivalent associates with normal autosomal bivalents (up to 18.2%). In single cells a premature desynapsis of sex chromosomes at late pachytene was observed. There was the appearance of single cells with the X0 karyotype. A 3-fold excess of the normal consumption of NA causes an increase in the number of cells with the above disturbance in the SC structure and behaviour and causes the appearance of heteromorphic bivalents. On the 7th day after a single MTD injection of NA male mice displayed a sharp (up to 27.9%) increase in the number of cells in which the sexual bivalent associated with autosomal bivalents, formation of univalents of autosomal and sex chromosomes, SC degeneration.

Adolescent↗

[A preparative method for isolating the synaptonemal complexes from mammalian spermatocytes].

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.

Animals↗

[Electron microscopic analysis of synaptonemal complexes of male laboratory mice exposed during the period of embryogenesis in the vicinity of the Chernobyl Nuclear Power Station].

Four male mice were shown to have decreased fertility among 74 laboratory male mice exposed in the Chernobyl APP area during embryogenesis. Electron microscopic analysis of synaptonemal complexes (SC) of at zygotene-diplotene and light-optic analysis of chromosome aberrations at diakinesis-metaphase 1 demonstrated the presence of interchromosome translocations in autosomal chromosomes, and also intrachromosome translocations in two of four animals. The frequency of chromosome translocations in SC preparations was on the average 1.8 times higher than their frequency at the diakinesis-metaphase 1 stage. High percentage of cells in which associations of sex bivalent axes (XY) with the axes of autosomal bivalents were observed in the SC preparations and reduced fertility in the animals studied confirmed the Forejt's hypothesis that such associations caused the arrest of cells at pachytene and, as a consequence, disturbed the process of embryogenesis.

Accidents↗

[Synaptonemal complexes of the A- and B-chromosomes of the spermatocytes from the East Asian mouse Apodemus peninsulae].

The analysis of whole-mount preparations of synaptonemal complexes (SCs) from surface-spread spermatocytes of A. peninsulae (2n = 48A + 1, 2, ... 12 B) had revealed SCs of 23 autosomal bivalents, sex bivalent XY, axial cores and SCs of the B-chromosomes. The intercellular and interindividual variability of the number of B-chromosomes varied from 1 to 12 per cell. The SCs of autosomal bivalents were shown to have a typical structure. The structure and behaviour of SCs of sex bivalent throughout meiotic prophase I appeared to be similar to those observed in other species of this order. Mainly B-univalents and less frequently B-bivalents containing SCs were found to be formed in meiotic prophase I. The full homologues appear to be rarely seen among B-chromosomes of the East-Asiatic mouse. A tendency of forming clusters of B-univalents near the sex bivalent was found, in addition to B-bivalents with lateral elements, having the form of bi- and tri-stranded elements with rare synaptic fragments. Besides this, the SCs of the autosomes of pachytene cells were found to contain structures resembling the recombination nodules.

Animals↗