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Biomedical subjects

T B Nesterova

Publications and source records attributed to T B Nesterova.

At least 37 records · Page 2Linked to original sources

[Prospects for obtaining a mapping panel for somatic cell marsupial-rodent hybrids for the short-tailed opossum Monodelphis domestica].

A possibility of obtaining a panel of marsupial-rodent somatic cell hybrid clones has been explored, with a view to mapping the genome of the opossum (Monodelphis domestica). Fusion of opossum cells (splenocytes, bone marrow cells, and fibroblasts) with fibroblasts of Chinese hamster or vole (HGPRT- and TK- mutants, respectively) produced 146 hybrid clones. The majority of marsupial-mammalian somatic cell hybrids were characterized by pronounced fragmentation and segregation of marsupial chromosomes. To overcome this difficulty, a method for rapid screening was developed, which allowed the early selection of clones rich in chromosomal material of opossum. Based on the screening results, 25 clones of independent origin were selected. A detailed genetic analysis, which included chromosome G-banding and in situ hybridization of biotin-labeled opossum DNA on metaphase chromosomes, allowed further selection of seven hybrid clones containing one to six intact chromosomes of M. domestica. Opossum chromosomes were present in various combinations against the background of Chinese hamster or vole chromosomes. The clones will be included in the panel of opossum-rodent somatic cell hybrids, which is currently being created.

Animals↗

[Rapid karyotyping of mammalian cells].

The use of "pipette" method ensures rapid preparation of standardized whole metaphase spreads. Experiments with human, murine, Chinese hamster, American mink, green African monkey, dog, and vole cells demonstrated that G-banded whole metaphase spreads can be obtained in less than two hours after the beginning of work with cell or tissue culture. Due to that, it became possible to start karyotyping of animal tissue explants, as well as fetal cells present in human amniotic fluid, on day 3 to 4 after their receiving.

Animals↗

[X chromosome inactivation in mammals].

The most important results of the last 30 years of studies on mammalian X-chromosome inactivation are reviewed. The data on X-chromosome inactivation in cells of embryonic and extraembryonic tissues and in male and female germ cell lines are discussed. Special attention is paid to data on mapping and functioning of the X-inactivation center and of recently discovered gene XIST. The main hypotheses concerning the mechanisms responsible for X-chromosome inactivation are considered. A new model of X-inactivation is proposed, which regards heterochromatin as a nonspecific activator of nucleation of the X-chromosome on which it is located.

Animals↗

[Construction of a mapping panel of human-rodent hybrid cells].

A clone panel of 27 human-Chinese hamster and 4 human-mouse somatic cell hybrids which contained as minimum five discriminating clones for any chromosome pairs was set up. Segregation analysis of 45 human chromosome-specific isoenzymes and PCR markers in hybrid clones allowed to demonstrate a possibility to apply the obtained panel for chromosome mapping of human genes.

Animals↗

[Establishment of linkage and the order of the genes GALA, G6PD, HPRT and PGK on the X-chromosome in two species of voles of the genus Microtus].

Localization of genes GALA, G6PD, HPRT and PGK on X-chromosome of Microtus subarvalis has been proved. Using the radiation hybrid mapping technique of Goss and Harris, the order of these genes for two species M. subarvalis and M. Kirgisorum was established. Statistical methods (program package RHMAP) result in the only gene order PKG--HPRT--G6PD--GALA for M. subarvalis. The same order was found to be the most probable for M. kirgisorum. Relative distances between these genes in two species appeared to be practically equal. A conservatism of a linear order of the X-linked genes in various mammalian taxons is discussed.

Animals↗

[Statistical validation of a radiation mapping method].

Correct statistical treatment of results of Goss and Harris experiments on radiation hybrid mapping is proposed. Using stochastic simulation a problem of an optimal sample size is investigated. Reasons for a repetition of experiments with different doses of radiation are discussed.

Animals↗

[The detection of the location of glucose-6-phosphate dehydrogenase in the fibroblasts of voles and mice and in rat myoblasts by using monoclonal antibodies against glucose-6-phosphate dehydrogenase].

Monoclonal antibodies (MAs) were produced against glucose-6-phosphate dehydrogenase (G6PD) of two vole species--Microtus arvalis and M. subarvalis. The binding level of the MAs to G6PD in both species were almost the same, which suggested that these MAs may be specific for the antigenic determinants common to G6PD of these species. The MAs produced against the vole G6PD were used for its intracellular localization. The patterns obtained after staining cells with the use of MAs against G6PD were the same as those obtained after staining with the use of antibodies against F-actin. There was a good conformity between the results of light and electron microscopic immunoenzyme analyses with regard to the binding of MAs produced to the actin microfilaments. It is concluded that G6PD is closely associated with actin microfilaments of the cell cytoskeleton.

Animals↗

[Comparative analysis of the level of heterozygosity for glucose phosphate isomerase (GPI) locus in silver foxes (Vulpes vulpes) of domesticated and control populations].

This communication is the first step of the studies on the correlation between protein polymorphism and the level of phenotypic diversity. The level of heterozygosity for glucose phosphate isomerase (GPI) locus was analysed in two populations of silver foxes. One of them has been selected for domestic behaviour during many years. This selection vector gave rise to many phenotypic novelties; 46% of the foxes analyzed has aberrant phenotypes. Another population was control bred under the same conditions of experimental farm of Siberian Dept. Russian Sci. with a commercial purpose. All the foxes analyzed from this population, except one, had a standard phenotype. Among 96 domestic foxes under analysis, only one heterozygotes for the GPI locus was detected. Among 112 control foxes, six were heterozygotes. In other words the data obtained indicate no correlation between the level of morphological diversity and the state of heterozygosity of the GPI locus.

Animals↗

[Structural heterochromatin and X-chromosome inactivation].

Our previous studies on the expression of the G6PD and alpha-GAL genes from the X chromosome of inter-specific hybrids of voles of the Microtus genus have demonstrated an unusual pattern of X-inactivation in the parents. The observed phenomenon was explained as the presumable result of nonrandom inactivation of the X chromosomes with a heterochromatin block in crosses involving Microtus arvalis whose X lacks a heterochromatin region and also of random X inactivation when both parents had heterochromatin blocks on the Xs. Based on known models, we discuss here the possible mechanisms of the effect of heterochromatin on X-inactivation; we give preference to the model postulating binding of nonhistone protein to the inactivation centre as the key event. The hypothesis we offer suggests change in chromatin conformation in the inactivation centre during packaging of heterochromatic region of a chromosome; the protein molecules diffusing along the chromosome towards the heterochromatin region by the "facilitated diffusion" mechanism may happen to be in the region of the X-inactivation centre, which, being in a favorable state, binds specifically to it; as a consequence, the binding probability of protein to heterochromatin increases as compared to chromosome without heterochromatin block.

Animals↗

Mapping of the silver fox genes: assignments of the genes for ME1, ADK, PP, PEPA, GSR, MPI, and GOT1.

Evidence is presented for the assignment of seven fox genes on the basis of the segregation data for chromosomes and enzymes of fox x Chinese hamster somatic cell hybrids. The chromosomal loci of the following enzyme genes were determined: ME1, VFU1; ADK and PP, VFU4; PEPA, VFU5; GSR, VFU7; and MPI and GOT1, VFU15. The localization of these genes now extends the fox genetic map to 22 mapped genes. Based on comparative analysis of mammalian genetic maps, karyotype evolution in Carnivora is discussed.

Animals↗

Mapping of silver fox genes: chromosomal localization of the genes for GOT2, AK1, ALDOC, ACP1, ITPA, PGP, and BLVR.

Evidence is presented for the chromosome localization of seven silver fox genes by the use of a panel of fox x Chinese hamster somatic cell hybrids. AK1, GOT2, and ALDOC are assigned to chromosome VFU2, PGP to chromosome VFU38, BLVR to chromosome VFU5, ACP1 to chromosome VFU8, and ITPA to chromosome VFU14. The genetic map of 29 fox genes is compared with those reported for man and other mammals. The results we obtained support and extend our previous suggestion that the formation of the Canidae branch of the Carnivora phylogenic tree was associated with a great increase in the rate of reorganization of the ancestral karyotype.

Acid Phosphatase↗

[Heterochromatin as a factor influencing X-chromosome inactivation in hybrids of the common vole (Microtinae, Rodentia)].

Expression of X-linked genes for G6PD and alpha-GAL was studied in female interspecific hybrids of Microtus. The G6PD and alpha-GAL isozymes of Microtus arvalis were found to predominate in all cases when a species carrying a heterochromatin block on the X-chromosome served as one partner of hybridization and M. arvalis containing no heterochromatin block served as another. The proportions of G6PD and alpha-GAL parental forms were approx. equal in hybrid females when both species participating in hybridization contained heterochromatin blocks on X-chromosomes. Cytological analysis for revealing active and nonactive X-chromosomes on metaphase spreads of hybrid females supports the biochemical data. Non-random inactivation of X-chromosomes carrying the heterochromatin blocks in the interspecific hybrids with M. arvalis and a random one, when both parents contain heterochromatin blocks on the X-chromosomes are supposed to be the cause for the phenomenon observed. The study provided data supporting our previous hypothesis that heterochromatin affects the X-chromosome inactivation process in interspecific hybrid voles.

Animals↗

[Mapping of the silver fox genome. III. Determination of the chromosomal localization of the GOT2, AK1, ALDOC, ACP1, ITPA, PGP and BLVR genes].

Evidence is presented for the chromosome localization of seven silver fox genes obtained with the help of panel of fox x Chinese hamster somatic cell hybrids. Thus, the AK1, GOT2 and ALDOC are assigned to chromosome VFU2, PGP to chromosome VFU3, BLVR to chromosome VFU5, ACP1 to chromosome VFU8 and ITPA to chromosome VFU14. The genetic map of 29 fox genes is compared with those reported for man and other animals. The results obtained support and extend our previous suggestion that formation of the Canidae branch of the Carnivora phylogenetic tree was associated with great increase in the rate of reorganization of the ancestral karyotype.

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[Genome mapping in silver fox. Syntenic genes in Carnivora].

Hamster X fox somatic cell hybrids segregating individual fox chromosomes in different combinations were used to assign seven structural loci to fox chromosomes. The gene for ME1 was mapped on the VFU1 chromosome, the genes for ADK and PP being located on the VFU4 chromosome. The gene for GSR was assigned to the VFU7 chromosome and the genes for MPI and COT1 were assigned to the VFU15 chromosome. Localization of these genes enhances the established fox genetic map and extends the known syntenic homologies between the fox and other mammalian. The comparison of data on gene mapping has provided basis for suggestion that there are significant differences in rates of karyotypic evolution in many mammalian taxa.

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Subchromosomal localization and order of GLA, PGK1, HPRT, and G6PD loci on the X chromosome of the American mink (Mustela vison).

Segregation of the X-linked mink markers alpha-galactosidase (GLA), phosphoglycerate kinase-1 (PGK1), hypoxanthine phosphoribosyltransferase (HPRT), and glucose-6-phosphate dehydrogenase (G6PD) was analyzed in hybrids of gamma-irradiated mink fibroblasts and Chinese hamster cells and in hybrids of nonirradiated mink fibroblasts and mouse hepatoma cells. Based on this analysis, the order of the four genes is GLA-PGK1-HPRT-G6PD on the mink X chromosome. Cytogenetic analysis of five mink x Chinese hamster hybrid clones containing mink GLA, PGK1, and HPRT, but lacking G6PD, tentatively localized mink G6PD to Xq15.22----qter and also confirmed the gene order as GLA-PGK1-HPRT-G6PD-qter. Comparison of this order with its counterpart in man and the mouse, as well as an analysis of the G-band patterns of their X chromosomes, demonstrated putative similarities between mink and man and differences in the mouse. These differences may be due to a different rate of X-chromosomal rearrangement in mammalian evolution.

Animals↗

Silver fox gene mapping: conserved chromosome regions in the order Carnivora.

Twenty-three silver fox x hamster somatic cell hybrid clones were used to assign 15 fox genes: GPI to chromosome 1; PGD to chromosome 2; MDH2 to chromosome 3; ESD to chromosome 6; LDHB to chromosome 8; NP to chromosome 10; LDHA to chromosome 11; APRT, ENO1, and PGM1 to chromosome 12; IDH1 and MDH1 to chromosome 16; and GLA, G6PD, and HPRT to the X chromosome. High-resolution G-banding of human, cat, mink, and fox chromosomes containing homologous regions (according to genetic maps) revealed regions of putative homology. The results lend support to the suggestion that the most considerable karyotypic reorganization of the ancestral genome in the order Carnivora occurred during Canidae formation. The details of karyotypic evolution in mammals are discussed.

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