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Effect of diabetes mellitus on reproductive parameters in mice
Margaryan, Hasmik ; Elzeinová, Fatima ; Kubátová, Alena ; Strolená, Eva ; Pěknicová, Jana
The decrease in population fertility has become a major concern in many developed countries. Recent studies show that infertility is affecting an estimated 15% of all couples (World Health Organization, WHO, 2010). Male infertility is the primary or contributing cause in 60% of cases. Male infertility is caused by a number of factors, such as genetic background, various environmental factors and disease. Diabetes mellitus (DM), a serious health problem on its own, is also suspected to be a contributing factor to male infertility. The aim of this project was to analyze the cellular, molecular and genetic effects of diabetic environment on spermatogenesis and sperm quality and to determine the impact of DM on the in vivo reproduction, using the mouse model (Mus musculus) inbred FVB. Diabetes was induced using streptozotocin. We used our knowledge and tools (unique monoclonal antibodies developed by our group) to determine the status of reproductive organs, anogenital distance, and the quality of sperms. Genetic analysis was performed by a quantitative Reverse Transcription Polymerase Chain Reaction (qPCR). We tested selected genes which are expressed in testicular tissue and thus can influence process of spermatogenesis and consequently the sperm quality. Our preliminary data strongly suggest that DM impairs male fertility. We have found significant changes in the body and reproductive organ weight of mice with DM. We have identified qualitative and quantitative changes in the expression of proteins in epididymal fluid and sperms. We have also detected an increased number of apoptotic cells in sperm of diabetic mice compared to the control group. To our knowledge, there is no study assessing the correlation between DM and “unexplained infertility”. In view of this, it is essential to analyze the effects of DM on male fertility, sperm quality, and reproduction parameters.
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Effect of exposure to bisphenol A on gene expression in the testicular tissue in male mice
Dorosh, Andriy ; Elzeinová, Fatima ; Žatecká, Eva ; Kubátová, Alena ; Pěknicová, Jana
Laboratory of Reproductive Biology, Institute of Biotechnology, Academy of Sciences of the Czech Republic, v.v.i., Prague, Czech Republic Bisphenol A (BPA) is a synthetic, endocrine-disrupting compound able to directly bind estrogen receptors. Free BPA has been detected in human samples indicating that humans are internally exposed to BPA. The purpose of this study was to analyse the effect of BPA on the male reproductive system and testicular gene expression in germ cells. We studied the influence of long-term low concentration BPA exposition on male fertility in vivo in a two-generation study in C57BL/6 mouse strain. In this work, BPA was added with water at two environmentally relevant concentrations: 0, 4 and 4 µg/l. We measured the reproductive organs weight and sperm cells morphology and quality. Expression of genes involved in endocrine regulation and energy metabolism in testis was analysed after BPA exposure. Next, the epigenetic mechanisms of gene expression and regulation during the germ cell differentiation and effect of BPA will be analysed.
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The effect of tetrabromobisphenol a on protamination and DNA quality of mouse sperm
Žatecká, Eva ; Castillo, J. ; Elzeinová, Fatima ; Kubátová, Alena ; Děd, Lukáš ; Pěknicová, Jana ; Oliva, R.
Tetrabromobisphenol (TBBPA) is a widely used brominated flame retardant, currently its consumption is 210,000 tons / year and is still growing. In our previous multigenerational in vivo study we have demonstrated that TBBPA is able to induce apoptosis of testicular cells and changes in the expression of genes important for proper spermatogenesis. However the potential effect of TBBPA on epidydimal spermatozoa had not yet been investigated. Therefore, we performed further study to evaluate the effect of on sperm DNA integrity and on the protamines as the major nuclear proteins. C57Bl/6J mice pups (n=10) were exposed to TBBPA (experimental group) during the gestation, lactation, pre-pubertal and pubertal periods up to the age of 70 days and compared control mice pups (n= 10) which were not exposed. Our results demonstrate that TBBPA treatment results in a significantly decreased P1/P2 ratio, increased total protamine/DNA ratio and increased DNA fragmentation observed between TBBPA and control mice, respectively. Protamines have recently been connected to the epigenetic marking of sperm chromatin in human and mouse spermatozoa. Thus, our findings suggest that TBBPA exposure, in addition to result in increased sperm DNA damage, may also alter the epigenetic marking of sperm chromatin.
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Flow cytometry (FCM) sperm assessment In normozoospermic and asthenozoospermic men using monoclonal antibodies against sperm proteins
Čapková, Jana ; Kubátová, Alena ; Děd, Lukáš ; Teplá, O. ; Pěknicová, Jana
Recent studies have shown that infertility affects an estimated 15% of all couples. Male infertility is the primary or contributing cause in 60% of these cases. Consequently, application of methods of assisted reproduction is increasing. These methods would benefit from extended evaluation of the sperm quality. For this purpose, we analyzed sperm proteins in men with normal spermiograms and with asthenozoospermia. Ejaculates of both groups were tested with a set of well-characterized monoclonal antibodies (MoAbs) to human sperm. No statistically significant differences were found between normospermics and asthenospermics in the expression of sperm surface proteins clusterin, evaluated by Hs-3 MoAb, and semenogelin, evaluated by Hs-9 MoAb. On the other hand, flow cytometry revealed quantitative differences between normozoospermic and asthenozoospermic men in GAPDHS (glyceraldehyde phosphate dehydrogenase human sperm-specific glycolytic enzyme), evaluated by Hs-8 MoAb, VCP (valosin-containing protein), detected with Hs-14 MoAb, and PRKAR2A (cAMP-dependent protein kinase type II – alpha regulatory subunit) detected by MoAb Hs-36. Asthenozoospermic men displayed significantly reduced expression of intra-acrosomal proteins with a likely decrease in sperm quality, and thus a negative impact on successful reproduction.
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Ekologická rozhraní: interakce živého a neživého v povrchových mikroekosystémech půd a antropogenních substrátů
Univerzita Karlova v Praze, Praha ; Peksa, Ondřej ; Vojta, Jaroslav ; Machová, Kateřina ; Škaloud, Pavel ; Prášil, Karel ; Kubátová, Alena ; Neustupa, Jiří ; Soldán, Zdeněk
Výsledky za rok jsou uvedeny v kapitolách: A. Abiotické a ekofyziologické charakteristiky studovaných lokalit, B. Studium sinic a řas, C. Studium hub, D. Studium lišejníků, E. Studium mechorostů, F. Inokulace substrátu sinicí rodu Nostoc. Uveden je přehled o čerpání finančních prostředků.
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Ekologická rozhraní: interakce živého a neživého v povrchových mikroekosystémech půd a antropogenních substrátů
Univerzita Karlova v Praze, Praha ; Mrázek, Jakub ; Vojta, Jaroslav ; Peksa, Ondřej ; Prášil, Karel ; Kubátová, Alena ; Škaloud, Pavel ; Machová, Kateřina ; Neustupa, Jiří ; Soldán, Zdeněk
V roce 2005 byl studován soubor dvanácti lokalit - tři přírodní stanoviště, pět odkališť, dvě výsypky a dvě kulturní plochy. Předběžné výsledky jednotlivých pracovních skupin: algologické, mykologické, bryologicko-lichenologické a geobotanické.
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Ekologická rozhraní: interakce živého a neživého v povrchových mikroekosystémech půd a antropogenních substrátů
Univerzita Karlova v Praze, Praha ; Peksa, Ondřej ; Vojta, Jaroslav ; Machová, Kateřina ; Škaloud, Pavel ; Prášil, Karel ; Kubátová, Alena ; Neustupa, Jiří ; Soldán, Zdeněk
Výsledky řešení projektu jsou uvedeny v kapitolách: 1. Abiotické a ekofyziologické charakteristiky studovaných lokalit, 2. Studium cyanobakterií a řas, 3. Studium hub, 4. Dekompozice listového opadu břízy a modelové celulózy, 5. Studium lišejníků, 6. Studium mechorostů, 7. Inokulace substrátu sinicí rodu Nostoc. V rámci jednotlivých kapitol jsou pak uvedeny i specifické metodiky. Návrh metodiky Monitoring a management antropogenních toxických substrátů rudních a strusko-popílkových odkališť bez souvislého vegetačního pokryvu.
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The effect of low dose of vinclozolin on reproductive tract development in CD1 outbred mice
Elzeinová, Fatima ; Pěknicová, Jana ; Nováková, Vendula ; Buckiová, Daniela ; Kubátová, Alena
The effect of a low dose of vinclozolin within the development of the reproductive tract during gestation (VIN-GD 15-22) and puberty (VIN-PND 23-44) in CD1 mice was tested. We found a decrease in the anogenital distance, prostate weight and pathology of testes in both experimental groups. Sperm counts decreased to 46% (VIN-GD) and to 81% (VIN-PND), and also the acrosomal state (evaluated by antiacrosomal antibody) decreased in both groups to 89% in comparison to the control group (100%). Sperm head abnormalities increased by approximately 18% and 13%, respectively. In this connection, the expression of some genes was changed (arosome-related gene (Acr), apoptosis related genes (p53, p21)). In conclusion, a low dose of vinclozolin affected the reproductive tract, sperm parameters and expression of selected genes in both experimental groups.
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