Volume 48 (2002) No. 4
Professor Morten Simonsen
J. Krejčí, K. Hála...........................................................................................125
Professor Vulchan Vulchanov
S. Marinova....................................................................................................125
Full text. 125
Review
Chicken Cells - Oncogene Transformation, Immortalization and More
J. PLACHÝ, J. HEJNAR........................................126
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jiří Plachý, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic. Tel.: +420 (205) 783827; Fax: +420 (2) 24310955; e-mail: plachy@img.cas.cz.
Abstract.
Full text. 126-128 129-130 131-133 134-136
Original Articles
Chromosome Assignment of Cd36 Transgenes in Two Rat SHR Lines by FISH and
Linkage Mapping of Transgenic Insert in the SHR-TG19 Line
F. LIŠKA1, G. LEVAN2, K. HELOU2, M. SLADKÁ1, M. PRAVENEC1, 3, V. ZÍDEK3, V. LANDA3, 4, V. KŘEN1,3......................................................139
1Institute of Biology and Medical Genetics, 1st Medical Faculty, Charles University, Prague
2Department of Cell and Molecular Biology and Genetics, Lundberg Laboratory, Goteborg
University, Gothenburg, Sweden
3Institute of Physiology, Academy of Sciences, Czech Republic
4Institute of Molecular Genetics, Academy of Sciences, Czech Republic
Corresponding author: Vladimír Křen, Institute of Biology and Medical Genetics, 1st Medical Faculty,
Charles University, Albertov 4, Prague 2, 128 00, Czech Republic. Tel.: +420 (2) 2496 8147; fax: +420 (2) 2491 8666; e-mail: vkren(zavináč)lf1.cuni.cz.
Abstract.
Full text. 139-140 141-142 143-144
Influence of Oviductal Fluid on Ovine Embryo Viability
M. LIBIK1, T. SLAVÍK2, T. SCHWARZ1, M. MURAWSKI1, S. CIURYK1,
E. WIERZCHOS1.................................145
1Department of Sheep and Goat Breeding, Agricultural University, Krakow, Poland
2Institute of Animal Physiology and Genetics, Academy of Sciences of the Czech Republic, Liběchov, Czech Republic
Corresponding author: Malgorzata Libik, Department of Sheep and Goat Breeding, University of Agriculture, al. Mickiewicza 24/28, Krakow, Poland, email: glibik@wp.pl.
Abstract.
Full text. 145-146 147-148
The Status of the Chromatin of Human Preimplantation Embryos with Good Morphology
S. M. DELIMITREVA............................149
Human IVF Laboratory, Department of Biology, Medical Faculty, Sofia, Bulgaria
Corresponding author: Stefka M. Delimitreva, Human IVF laboratory, Department of Biology, 1,
St. George Sofiisky Street, 1431 Sofia, Bulgaria.
Abstract.
Full text. 149-150 151-153
Short Communications
Conditions for Gene Transfection into the HL-60 Human Leukaemia Cell Line by Electroporation
J. PACHERNÍK, R. JANÍK, J. HOFMANOVÁ, V. BRYJA, A. KOZUBÍK..................154
Laboratory of Cytokinetics and Flow Cytometry, Institute of Biophysics, Academy of Sciences of the Czech Republic, Brno, Czech Republic
Corresponding author: Alois Kozubík, Institute of Biophysics, Academy of Sciences of the Czech Republic, 612 65 Brno, Královopolská 135, Czech Republic. Tel.: +420-5-41517182; fax: +420-5-41211293; e-mail: kozubik@ibp.cz.
Abstract.
Full text. 154-155 156
Genetic Determination of Polyhydroxyalkanoate Metabolism in Rhodobacter capsulatus SB1003
P. ULBRICH1, H. STRNAD1, V. HEJKALOVÁ1, J. PAČES2, V. PAČES2 .....................157
1Department of Biochemistry and Microbiology, Institute of Chemical Technology, Prague, Czech Republic
2Department of Genomics and Bioinformatics, Institute of Molecular Genetics, Academy of Sciences, Prague, Czech Republic.
Corresponding author: Václav Pačes, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 16637 Praha 6, Czech Republic. Tel.: +42 (02) 20183 541; Fax: +42 (02) 2431 0955; e-mail: vpaces@img.cas.cz.
Abstract.
Full text. 157-159
Asymmetric Vimentin Distribution in Human Spermatozoa
M. D. MARKOVA, T. T. MARINOVA, I. T. VATEV......................................160
Department of Biology, Medical Faculty, Medical University of Sofia, Bulgaria
Corresponding author: Maya D. Markova, Department of Biology, Medical Faculty, 2 Zdrave Street, BG-1431 Sofia, Bulgaria. E-mail: mmarkova@medfac.acad.bg.
Abstract.
Full text. 160-161 162
Book Reviews
Pavel Klener: Clinical Oncology
V. PAČES..................................................................163
Full text. 163-164
Review
Chicken Cells - Oncogene Transformation, Immortalization and More
J. PLACHÝ, J. HEJNAR
Abstract. Domestic chicken as a laboratory animal as well as chicken cells in vitro have been highly evaluated in several fields of experimental biology. Retrovirology and experimental oncology traditionally use this model, whose comparative aspects are still inspirative. The first (retro)viral aetiology of a tumour was recognized in the chicken and the first quantitative in vitro measurement of oncogenic transformation was developed using the chicken cells. Chicken cells (like human and primate, but unlike rodent cells) have a long primary life-span, during which they remain genetically stable. While this property is advantageous for several types of experiments, it correlates with a low propensity of the chicken cells to immortalization. Recent establishment of several continuous chicken cell lines, however, has surmounted this drawback. Furthermore, the chicken B cell line DT40 was proved to be extremely useful for gene disruption studies because of a high frequency of gene targeting not found in any vertebrate cells. In the present communication, we have tried to review several traditional achievements accomplished using the chicken model and point to newly opened areas, where chicken cells appear to be an efficient tool, particularly in cell transformation and immortalization.
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Chromosome Assignment of Cd36 Transgenes in Two Rat SHR Lines by FISH and
Linkage Mapping of Transgenic Insert in the SHR-TG19 Line
F. LIŠKA, G. LEVAN, K. HELOU, M. SLADKÁ, M. PRAVENEC, V. ZÍDEK, V. LANDA, V. KŘEN
Abstract. The chromosome position of the Cd36 insert was determined by FISH in two rat transgenic lines (SHR/Ola-TgN(EF1aCd36)10Ipcv (SHR-TG10) and SHR/Ola-TgN(EF1aCd36)19Ipcv (SHR-TG19). The Cd36 transgene construct labelled with digoxigenin-11-dNTP was used as a probe in the FISH analysis. In accord with the previous finding that the SHR-TG10 harbours 6–8 copies of the transgene, the signals from both metaphase and interphase nuclei of SHR-TG10 preparations were rather strong and the probe hybridized to both copies of chromosome 1 at band q55. The probe hybridization to SHR-TG19 metaphase preparations also showed homozygosity of the transgene with localization of both copies to chromosome 11 at band q11. The signals were distinct but much weaker compared to the SHR-TG10, which again is in accord with the fact that the SHR-TG19 line harbours only a single copy of the transgene. In order to look for a possible impact of the insertion site neighbourhood upon the transgene phenotypic effect, we performed linkage mapping of the transgene in the SHR-TG19 line. By linkage mapping, the placement of the transgene to the proximal part of RNO11 was confirmed, the critical interval being 4 cM between D11Rat20 and D11Rat21, in good agreement with the RH map. Within the close neighbourhood of the inserted Cd36 transgene, there are several genes known to be expressed in kidney, and so the influence of some regulatory sequences enhancing kidney expression of the Cd36 transgene can be envisaged.
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Influence of Oviductal Fluid on Ovine Embryo Viability
M. LIBIK, T. SLAVÍK, T. SCHWARZ, M. MURAWSKI, S. CIURYK,
E. WIERZCHOS
Abstract. The severe loss of developmental competence affecting fertilized ova when removed from the oviductal environment suggests that this organ plays a functional role in early embryonic development. The purpose of this study was to determine the effect of sheep heat-inactivated OF on the mortality rate of ovine embryos produced in vitro and transferred into recipients. As control groups we used embryos fertilized and cultured in media supplemented with different kinds of proteins (FCS, BSA). Transfer of embryos in the two pronuclei stage to the oviducts of synchronized recipients resulted in 60% of successfully termed pregnancies after incubation of embryos in OF, 40% in BSA and only 10% after FCS. All ewes were further assessed for pregnancy by ultrasonography 33, 53 and 80 days after embryo transfer. The highest embryo mortality appeared between day 33 and 52. We concluded that incubation of ovine oocytes in OF during the final period of the maturation process may play a functional role at the time of fertilization and early embryonic development.
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S. M. DELIMITREVA
Abstract. The data about the relation and succession of blastomere fragmentation, cleavage rate and chromatin anomalies in preimplantation mammalian embryos are empirical and controversial at present. In this work we studied the proportion of nuclear fragmentation and condensation in 3–5-cell stage human embryos with no or minimal blastomere fragmentation (morphological class A and B, respectively) and the possibilities to perform FISH chromosomal analyses with them. We observed different stages of chromatin damage in blastomere nuclei corresponding to the steps of nuclear apoptotic changes well known in many cell types. The ploidity analysis of chromosomes 1, 5, 19 and X was determined as successful in embryos which had at least 2 out of 3, 3 out of 4 or 3 out of 5 normal nuclei with an equal number of FISH signals. There was no difference in the percentage of abnormal nuclei among the A- and B-class embryos. Tendencies noted by us suggest that the minimal blastomere fragmentation (up to 20% of perivitelline space) does not preclude the normal nuclear status allowing successful ploidy testing. The presence of condensed chromatin is a critical factor for interphase cytogenetic analysis of single early blastomeres.
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Short Communications
Abstract. Electroporation represents a powerful technique for cell transfection; however, its efficiency in haemopoietic cells (~1%) is largely unsatisfactory. Biological processes in haemopoietic cells are often studied using leukaemia cell line HL-60. For this reason we developed conditions for efficiently introducing plasmids to HL-60 cells by electroporation, as an alternative to other techniques. This technique employs the electric pulse (250–270 V; 1000 microF) followed by separation of living cells on a Ficoll-Paque discontinuous gradient. Using 10–20 microg of plasmid, we routinely achieve 12–14% of transfectants.
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Genetic Determination of Polyhydroxyalkanoate Metabolism in Rhodobacter capsulatus SB1003
Abstract. A cluster of four genes was identified in the Rhodobacter capsulatus genome that is involved in PHA metabolism. These genes encode the PHA granule-associated protein (pha2), the regulator for granule formation (pha1), the PHA synthase (phaC) and the PHA depolymerase (orfX). Two other genes, namely those encoding b-ketothiolase (phaA) and acetoacetyl-CoA reductase (phaB), are not linked to this cluster.
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M. D. MARKOVA, T. T. MARINOVA, I. T. VATEV
Abstract. The subject of the study was vimentin distribution in spermatozoa from human ejaculates by immunofluorescence and immunogold electron microscopy. In accordance with earlier reports, vimentin was found in the sperm head and was localized mainly in the equatorial segment region. However, electron microscopic observations revealed an additional intriguing detail: vimentin-associated gold granules showed asymmetric distribution. This asymmetry tended to be more pronounced in heads with defects such as cytoplasmic droplets. Abnormal cells also had positive reaction for vimentin in the neck and the proximal midpiece, but in these domains gold granule distribution was apparently uniform. These findings seem to support the hypothesis that the surfaces of the mammalian sperm head are functionally non-equivalent, although morphological basis for such a phenomenon is evident only in rodents. It is also interesting that asymmetry in vimentin distribution correlates with distinct types of sperm structural defects.
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