Volume 46 (2000) No. 1

Editorial
New Resources of Functional Genomics: Unique Vertebrate Models from Prague.
J. FOREJT.......................................................1
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jiří Forejt, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague 4, Czech Republic. Tel.: (420 2) 4752257; fax: (420 2) 4713445; e-mail: jforejt@biomed.cas.cz.
No abstract available.
Full text pp. 1-2

Articles
Depression in the Level of Cadherin and alfa-, beta-, gamma-catenins in Transgenic Xenopus laevis Tissues Highly Expressing c-Src
K. DVOŘÁKOVÁ¹, V. HABROVÁ¹, M. TAKÁČ², J. JONÁK²...........................3
¹Department of Physiology and Developmental Biology, Faculty of Science, Charles University, Prague, Czech Republic
²Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jiří Jonák, Department of Protein Biosynthesis, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo náměstí 2, 16637 Prague 6, Czech Republic; e-mail: jjon@img.cas.cz.
Abstract.
Full text pp. 3-5 6 7 8-9

Irradiation of Genetically Modified Plasmacytoma Vaccines Results in Upregulation of CD80 Molecule Expression, IL-2 Production and Higher Therapeutic Efficacy of the Vaccines
ŠÍMOVÁ J., REINIŠ M., SOBOTA V., ČAPKOVÁ J., BUBENÍK J., JANDLOVÁ T...........11
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jana Šímová, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 37 Prague 6.
Abstract.
Full text pp. 11-16

The Chicken - a Laboratory Animal of the Class Aves
J. PLACHÝ...............................17
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.
Abstract.
Full text pp. 17-24

SHR.BN-Congenic Strains for Genetic Analysis of Multifactorially Determined Traits
V. KŘEN^1,2, D. KŘENOVÁ¹, M. ŠIMÁKOVÁ², A. MUSILOVÁ², V. ZÍDEK²,
M. PRAVENEC^1,2.........................................25
¹Institute of Biology and Medical Genetics, 1^st Medical Faculty, Charles University, Prague, Czech Republic
²Institute of Physiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Michal Pravenec, Institute of Physiology, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague 4, Czech Republic. Telephone/Fax: (4202) 475 2297; e-mail:
pravenec@biomed.cas.cz.
Abstract.
Full text pp. 25-30

PWD/Ph and PWK/Ph Inbred Mouse Strains of Mus m. musculus Subspecies - a Valuable Resource of Phenotypic Variations and Genomic Polymorphisms
S. GREGOROVÁ, J. FOREJT..............................................31
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jiří Forejt, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague 4, Czech Republic. Tel. (420 2) 4752257; Fax (420 2) 4713445; e-mail: jforejt@biomed.cas.cz.
Abstract.
Full text pp. 31-33 34-37 38-41

Colon Mucosal Cells after High-Dose Fractional Irradiation
R. ZORC-PLESKOVIČ, O. VRASPIR-PORENTA, D. PETROVIČ, M. ZORC,
L. PLESKOVIČ*..............................................43
Institute of Histology and Embryology, Medical Faculty, University of Ljubljana, Ljubljana, Slovenia
* Department of Gastroenterologic Surgery, University Medical Centre of Ljubljana, Ljubljana, Slovenia
Corresponding author: Ruda Zorc-Pleskovič, Institute of Histology and Embryology, Medical Faculty of Ljubljana, Korytkova 2, 1105 Ljubljana, Slovenia. Tel.: +386 61 441 121; fax: +386 61 1401 294; e-mail: daniel.petrovic@mf.uni-lj.si.
Abstract.
Full text pp. 43-44 45 46-48

Development of the Ectopia Cordis Induced by Hydrocortisone Administration
V. SEICHERT¹, L. HERINGOVÁ², A. SEICHERTOVÁ², R. JELÍNEK².....................49
¹Institute of Anatomy, 1^st Faculty of Medicine, ²Centre of Biomedical Sciences, 3^rd Faculty of Medicine, Charles University, Prague, Czech Republic
Coresponding author: Václav Seichert, Institute of Anatomy, 1st Faculty of Medicine, Charles University, Prague 2, Czech Republic. Tel.: +420 (2) 24915003, Fax.: +420 (2) 297692.
Abstract.
Full text pp. 49-50 51 52 53-54

Short Communication
Monoclonal Antibody to Human Sperm Acrosomal Protein
J. ČAPKOVÁ, G. GEUSSOVÁ, J. PĚKNICOVÁ.....................................55
Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Prague, Czech Republic
Corresponding author: Jana Čapková, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20 Prague 4, Czech Republic. Fax: 420 2 444 717 07; E-mail: capkova@biomed.cas.cz.
Abstract.
Full text pp. 55-57

Articles
Depression in the Level of Cadherin and alfa-, beta-, gamma-catenins in Transgenic Xenopus laevis Tissues Highly Expressing c-Src
K. DVOŘÁKOVÁ, V. HABROVÁ, M. TAKÁČ, J. JONÁK

Aberrant morphogenesis of transgenic Xenopus laevis 5-day embryos carrying Rous sarcoma virus LTR in their DNA and expressing high level of c-Src protein kinase was found to be accompanied with a profound depression in the level of cadherins and alfa-, beta-, and gamma-(plakoglobin) catenins in their tissues, as revealed by immunohistochemical analysis. Simultaneously, an increased level of phosphotyrosine staining was detected. However, an analogous increase in the level of phosphotyrosine immunostaining and a slightly higher level of Src were also detected in tissues of originally defective but later spontaneously repaired frog embryos that displayed essentially normal patterns of staining for cadherins and catenins. Our results provide evidence that the defective morphogenesis of frog embryos expressing a high level of c-Src is characterized by the loss of the cadherin-catenin complexes. It appears that to induce frog morphogenetic malformations, the c-Src overproduction and the loss of cadherins-catenins are simultaneously required. Phosphorylation is not likely to be the cause of cadherin and catenin disappearance from the tissues of strongly aberrant frog embryos.
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It has been found previously that irradiated, IL-2 gene-modified plasmacytoma (X63-m-IL-2) vaccines are more efficient in the therapy of the parental (X63-Ag8.653) plasmacytoma than live plasmacytoma vaccines. In this communication, we have demonstrated that irradiation of murine IL-2-producing plasmacytoma vaccines resulted in upregulation of CD80 molecule expression and IL-2 production. The expression of MHC class I antigens was not altered. The upregulation of CD80 membrane molecule expression in X63-m-IL-2 cells was higher after irradiation with 150 Gy than after irradiation 50 Gy. Comparable upregulation of the CD80 molecule expression has also been demonstrated after irradiation of the parental murine X63-Ag8.653 plasmacytoma cells. The results indicate that upregulation of the CD80 molecule expression and enhanced IL-2 production in irradiated X63-m-IL-2 cells was responsible for their higher therapeutic effectiveness of the irradiated plasmacytoma vaccine.
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The Chicken - a Laboratory Animal of the Class Aves
J. PLACHÝ

Prague inbred lines of chickens represent a unique system of MHC(B) congenic partners differing in the immune-based resistance/susceptibility to v-src-induced oncogenesis. Mapping in chickens can be facilitated by the availability of inbred lines, since many well described differences in disease susceptibility and MHC(B) haplotypes exist among the defined lines. Long-term intensive research on human, mouse, and rat MHC has established a canonical picture of this multigene complex. The chicken MHC(B) is clearly the best characterized outside the mammals and it was the first MHC clearly different from the paradigmatic structure of the above mentioned mammalian species. Chickens were in many aspects the poor relatives of mice, and they had to wait for introduction of molecular biology methods. But, when it happened, the newly gained data could be easily reconciled with classical genetic studies using available congenic chicken lines. We have established permanent tumor cell lines from ex vivo tumors induced by the LTR, v-src, LTR provirus in inbred chickens. These cells express a high level of the v-src oncogene and are of defined MHC(B) genotype. We witness a dramatic acceleration of the development of chicken (avian) genomics. The chicken is not only a good comparative model for basic science, but it is also an object of the poultry industry, which is threatened by several avian diseases. The reason for genome mapping in chickens is thus more than academic.
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SHR.BN-Congenic Strains for Genetic Analysis of Multifactorially Determined Traits
V. KŘEN, D. KŘENOVÁ, M. ŠIMÁKOVÁ, A. MUSILOVÁ, V. ZÍDEK,
M. PRAVENEC

The laboratory rat is an important laboratory animal with multiple well-defined inbred strains, including some of the most widely used animal models of human diseases. Recent advances in the development of rat genetic resources will enable the exploitation of the full potential of rat models of human diseases and, in addition, the rat can provide useful information for comparative genomics of humans and mice. In the current review, we describe the development of a congenic strain by introgression of differential chromosome segments from the Brown Norway (BN) rat to the genetic background of the spontaneously hypertensive rat (SHR). These SHR.BN-congenic strains and recombinant sublines derived from them were developed as a model system for genetic analysis of multifactorially determined pathophysiological and morphological conditions.
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PWD/Ph and PWK/Ph Inbred Mouse Strains of Mus m. musculus Subspecies - a Valuable Resource of Phenotypic Variations and Genomic Polymorphisms
S. GREGOROVÁ, J. FOREJT

PWD/Ph and PWK/Ph (abbreviated PW*) are highly inbred mouse strains (F66 and F70) derived from wild mice of Mus musculus musculus subspecies. When compared with laboratory inbred strains, they display a plethora of differences in many complex phenotypes such as body weight, fat distribution pattern, blood levels of intermediate metabolites, sensitivity to type-1 diabetes or behaviour patterns. The PWD/Ph genes can rescue the lethal effect of lack of the Igf2 receptor. The male-limited hybrid sterility of (PWD/Ph x laboratory strain)F1 hybrids is a specific phenotype controlled by three or four unlinked loci. These complex phenotypic traits can be genetically dissected by QTL analysis using microsatellite markers of known genetic location. The PW* strains are particularly useful for such genome-wide scans since 70-80% of randomly chosen microsatellite markers are polymorphic in (PW* x laboratory strain) crosses compared to 35-45% in crosses between two laboratory strains. The list of polymorphic microsatellite loci is included in this report. The high degree of sequence polymorphism allows easier distinction between paternal and maternal mRNA transcripts in PW* hybrids, which makes the PW* strains a useful tool also in molecular studies of genomic imprinting. The high frequency of phenotypic differences together with the high degree of sequence polymorphism and the relatively easy breeding of PW* strains make them a valuable mammalian model organism for the functional genomics of the traits of biomedical importance.
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Colon Mucosal Cells after High-Dose Fractional Irradiation
R. ZORC-PLESKOVIČ, O. VRASPIR-PORENTA, D. PETROVIČ, M. ZORC, L. PLESKOVIČ

The aim of this study was to investigate histological and stereological changes in cryptal enterocytes, mucosal lymphocytes and mast cells 10 days after irradiation. For experimental model, 24 Beagle dogs 1-2 years old were used. Twelve dogs were irradiated 20 days with 32 Gy over the whole pelvis and tail. Another 12 dogs represented a control group. For the detection of apoptosis, the TUNEL technique was used. Histological and stereological analyses were performed using a Wild sampling microscope M 1000. In the irradiated group, volume density (P < 0.01), numerical density (P < 0.05) and average volume of lymphocytes (P < 0.001) were significantly lower than in the nonirradiated group. Numerical areal density of mast cells in the irradiated group was also significantly lower (P < 0.05). Volume density (P < 0.001) and average volume of mast cells (P < 0.001) were significantly higher in the irradiated group. The results of our experiments show that irradiation causes injury and loss of lymphocytes and mast cells in the colon mucosa. Apoptosis was detected in enterocytes and lymphocytes in the irradiated group and in nonirradiated group in equal numbers (2.5+/- 0.3 vs. 2.3 +/- 0.3; ns.), suggesting that 10 days after high-dose irradiation, the cell loss is not due to apoptosis.
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Development of the Ectopia Cordis Induced by Hydrocortisone Administration
V. SEICHERT, L. HERINGOVÁ, A. SEICHERTOVÁ, R. JELÍNEK

Our previous study on the development of thorax in chick embryos revealed that mechanical disturbance of the so-called membrana reuniens causes the development of the ectopia cordis (EC). To assess whether membrana reuniens disturbance was really essential for EC development, we employed hydrocortisone, a teratogen known to produce a high incidence of EC. The incidence of EC after the hydrocortisone intraamniotic application on the 4th embryonic day reached 84.8%. It was found that although in the whole course of EC development membrana reuniens appeared very thin, it nevertheless remained continuous. The morphology of membrana reuniens in embryos with fully developed EC, studied in classical serial histological sections, was similar to the amniotic membrane. Flow cytometry analysis of the cell cycle revealed that EC induced by hydrocortisone administration was associated with a significantly lowered proliferation activity of the prospective body-wall mesenchyme involved in the closure of the anterior wall of thorax. The probable mechanism of EC development is suggested.
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Short Communication
Monoclonal Antibody to Human Sperm Acrosomal Protein
J. ČAPKOVÁ, G. GEUSSOVÁ, J. PĚKNICOVÁ

A new monoclonal antibody designated Hs-14 was generated after immunization of BALB/c mice with the acid extract of human sperm. In indirect immunofluorescence Hs-14 mAb binds to the acrosome of permeabilized sperm cells and consequently recognizes some intra-acrosomal protein. Western blotting analysis revealed that under non-reducing conditions the Hs-14 mAb detects a protein with a molecular mass of 220 kDa. Under reducing conditions the Hs-14 recognizes several peptide bands within the range from 55 kDa to 110 kDa. Beside human sperm the antibody positively reacts also with sperm of some other mammalian species. Using Hs-14 mAb it is possible to evaluate the acrosomal integrity of spermatozoa and to reveal sperm pathology.
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