Volume 44/1998 No. 5

Volume 44/1998 No. 5
Articles
Linkage Mapping of the Fos Cellular Oncogene (Fos) to Rat Chromosome 6 and Its Possible Role in the Regulation of Compensatory Renal Growth
M. PRAVENEC1,2, V. ZÍDEK1, M. ŠIMÁKOVÁ1, J. VORLÍÈEK1, V. KØEN1,2
.........................151
1Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
2Institute of Biology, 1st Medical Faculty, Charles University, Prague , Czech Republic Corresponding author: Michal Pravenec, Institute of Physiology, Academy of Sciences of the Czech Republic, Vídeòská 1083, 142 20 Prague 4, Czech Republic. Tel/Fax: (4202) 475 2297; e-mail pravenec@biomed.cas.cz.
Abstract.
Angiogenesis of Extra- and Intraembryonic Blood Vessels is Associated with Expression of Nestin in Endothelial Cells
J. MOKRÝ, S. NÌMEÈEK ...........155
Department of Histology and Embryology, Medical Faculty, Charles University, Hradec Králové, Czech Republic
Corresponding author: Jaroslav Mokrý, Department of Histology and Embryology, Medical Faculty, Charles University, Šimkova 870, 500 01 Hradec Králové, Czech Republic. Fax +420 49 5513 597; Tel. +420 49 5816 471; e-maimokry@llfhk.cuni.cz.
Abstract.
The pyrAb Gene Coding for the Large Subunit of Carbamoylphosphate Synthetase from Bacillus stearothermophilus: Molecular Cloning and Functional Characterization
H. VLÁŠKOVÁ, L. KRÁSNÝ, V. FUÈÍK, J. JONÁK......163
Department of Protein Biosynthesis, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jiøí Jonák, Department of Protein Biosynthesis, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6, Czech Republic.
Abstract.
The Effect of Irradiation on FRTL-5 Cell Morphology
D. ŠTIBLAR-MARTINÈIÈ, A. CÖR.......173
Institute of Histology and Embryology, Medical Faculty, University of Ljubljana, Ljubljana, Slovenia
Corresponding author: Draga Štiblar-Martinèiè, Institute of Histology and Embryology, Medical Faculty, University of Ljubljana, 1105 Ljubljana, Slovenia. Tel. +386 61 14 03 042; Fax +386 61 14 01 294; e-mail draga.stiblar@mf.uni-lj.si.
Abstract.
Interaction of Bull, Stallion and Boar Seminal Plasma Proteins and Sperms with Acidic Polysaccharides
J. LIBERDA1, M. TICHÁ1, Z. ZRALÝ2, D. ŠVECOVÁ2, Z. VÌŽNÍK2
.........................177
1Department of Biochemistry, Faculty of Natural Science, Charles University, Prague, Czech Republic
2Veterinary Research Institute, Brno, Czech Republic
Corresponding author: Marie Tichá, Department of Biochemistry, Faculty of Natural Science, Charles University, 128 40 Prague 2, Albertov 2030, Czech Republic. Tel. 420 2 2195 2330. Fax 420 2 2195 2331.
Abstract.
Short Communication
Blasticidin Resistance Cassette in Symmetrical Polylinkers for Insertional Inactivaton of Genes in Dictyostelium
F. PÙTA1, C. J. ZENG2 .........................185
1Department of Physiology and Developmental Biology, Faculty of Science, Charles University, Prague, Czech Republic
2Department of Genetics, University of Kassel, Kassel, Germany
Corresponding author: František Pùta, Department of Physiology and Developmental Biology, Faculty of Science, Charles University, 128 44 Prague 4, Vinièná 7, Czech Republic.
Abstract.
Articles
Linkage Mapping of the Fos Cellular Oncogene (Fos) to Rat Chromosome 6 and Its Possible Role in the Regulation of Compensatory Renal Growth
M. PRAVENEC1,2, V. ZÍDEK1, M. ŠIMÁKOVÁ1, J. VORLÍÈEK1, V. KØEN1,2
Using the analysis of HXB/BXH recombinant inbred (RI) strains, we have recently detected a suggestive association between the D6Mit9 marker and a compensatory growth of the remnant kidney after unilateral nephrectomy (P = 6 x 10-4). Based on conserved synteny between mice, rats, and humans, the putative quantitative trait locus (QTL) might map to a region with several positional candidate genes: for instance, the Fos oncogene was implicated as a possible candidate gene for compensatory renal growth (CRG). In the current study, we mapped the Fos gene in RI strains 27 cM apart from the position where the putative CRG regulatory QTL was detected. Despite relatively wide confidence intervals for mapping QTLs with modest effects in a small population of RI strains, this finding probably excludes the Fos as a positional candidate for CRG.
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Angiogenesis of Extra- and Intraembryonic Blood Vessels is Associated with Expression of Nestin in Endothelial Cells
J. MOKRÝ, S. NÌMEÈEK
For a long time, intermediate filament protein nestin was recognized as a specific constituent of the cytoskeleton of developing neural cells and skeletal muscle. Recent reports by other investigators demonstrated that nestin may also be expressed in other cell types. Here we give evidence that nestin is expressed by endothelial cells of developing blood vessels in the rat. Using anti-Rat-401 monoclonal antibody we identified high levels of nestin in the endothelium lining all blood vessels of E14-15 rat foetuses. Immunoreactivity for nestin was detected in both extraembryonic (chorion, placenta, umbilical cord) and intraembryonic blood vessels. In the body of the foetus, we observed nestin-positive endothelial cells in vessels located in the areas of the mesenchyme as well as in vessels supplying the developing organs (central nervous system, liver, lung, spleen, heart, digestive tube). The strongest positive signal was given by the endothelium of newly vascularized tissues. In contrast to the developing tissues, expression of nestin by vascular endothelial cells was greatly reduced in adult tissues.
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The pyrAb Gene Coding for the Large Subunit of Carbamoylphosphate Synthetase from Bacillus stearothermophilus: Molecular Cloning and Functional Characterization
H. VLÁŠKOVÁ, L. KRÁSNÝ, V. FUÈÍK, J. JONÁK
The Bacillus stearothermophilus pyrAb gene, encoding the large subunit of carbamoylphosphate synthetase, was isolated and characterized. The DNA sequence is a 3195-nucleotide long reading frame coding for a polypeptide of 1064 amino acids and deduced Mr 116,160 Da. The pyrAb gene is part of the B. stearothermophilus pyrimidine biosynthesis operon pyr. The 5' end of the pyrAb gene overlaps with the 3' end of the pyrAa gene coding for the small subunit of carbamoylphosphate synthetase, while the 3' end of the pyrAb gene is overlapped with the 5' end of the pyrD gene, which by analogy with the B. subtilis genomic organization, codes for dihydroorotate dehydrogenase. The deduced amino acid sequence of the large subunit of B. stearothermophilus carbamoylphosphate synthetase was compared with the known sequences of the large subunit of carbamoylphosphate synthetase enzymes from other organisms via the NCBI database. Extremely high (98%) identity in amino acid sequence with the large subunit of carbamoylphosphate synthetase from Bacillus caldolyticus was detected.
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The Effect of Irradiation on FRTL-5 Cell Morphology
D. ŠTIBLAR-MARTINÈIÈ, A. CÖR
The morphological changes and proliferative activity of FRTL-5 cells irradiated in vitro with one dose of 0, 8, 12, 16, 24 or 32 Gy were studied morphometrically 7, 14 and 21 days after irradiation. The number of cells irradiated with 24 Gy and 32 Gy decreased during the 3 weeks of the experiment. Morphometrical analysis confirmed that the cells repaired their injuries up to 16 Gy of irradiation, while doses of 24 and 32 Gy damaged them irreversibly. Thyroglobulin was detected in cells after all doses of irradiation.
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Interaction of Bull, Stallion and Boar Seminal Plasma Proteins and Sperms with Acidic Polysaccharides
J. LIBERDA1, M. TICHÁ1, Z. ZRALÝ2, D. ŠVECOVÁ2, Z. VÌŽNÍK2
The interaction of seminal plasma proteins, sperms and detergent-released sperm proteins of three species with different types of acidic polysaccharides was studied. Heparin-binding activity of boar, bull and stallion seminal plasma proteins, sperms and sperm proteins was compared with their ability to interact with polysaccharides differing in the presence of the sulfate groups or in their saccharide moiety (chondroitin sulfate, dextran sulfate, fucoidan, hyaluronic acid). Bull seminal plasma proteins were characterized by higher affinity to heparin, fucoidan and dextran sulfate, while significant differences between different types of polysaccharides were detected in the case of boar proteins. Sperm protein interactions with acidic polysaccharides in bull and stallion were analogous to the binding of seminal plasma proteins. Bull and stallion seminal plasma proteins inhibited the interaction of corresponding sperm proteins with acidic polysaccharides.
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Short Communication
Blasticidin Resistance Cassette in Symmetrical Polylinkers for Insertional Inactivaton of Genes in Dictyostelium
F. PÙTA1, C. J. ZENG2
In Dictyostelium discoideum inactivation of developmentally regulated genes via homologous recombination has become an important tool in studying systematically the entire developmental program of this model organism. The Dictyostelium genome is very A/T-rich, but presents obstacles to the preparation of knockout constructs. The coding regions offer few suitable restriction sites and the low complexity intergenic regions do not guarantee specificity of recombination. We present here the preparation of plasmids pBsR479, pBsR503, and pBsR519, in which a blasticidin resistance-cassette is positioned in the center of various symmetrical polylinkers. This design simplifies the cloning process and gives more flexibility in positioning the selectable marker within the coding regions.
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