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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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METHODICAL RECOMMENDATION FOR INSTITUTES OF THE AS CR CONCERNING REPROTING OF REAL ELIGIBLE INDIRECT COSTS OF THE PROJECT
Škoda, Jan ; Starosta, Karel
The aim of the Methodical Recommendation is to create possible common principles for institutes of the Academy of Sciences of the Czech Republic (AS CR) concerning the allocation of indirect costs to projects, considering mainly the rules of fund providers requesting reporting of real costs via full cost methodology. Proposed methodology is based on usual management and accounting practices of institutes of AS CR and lie in principles of maximum simplicity and effectiveness. It should result in minimum increase of real indirect costs caused by application of such methodology, it enables closing of economic year on time and also reporting of necessary data to fund providers.
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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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The effect of low dose of vinclozolin on reproductive tract development in CD1 outbred mice
Pěknicová, Jana ; Elzeinová, Fatima ; 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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Povrchové proteiny spermie: původ, biochemické vlastnosti a úloha v reprodukci
Jonáková, Věra ; Postlerová, Pavla ; Tichá, M. ; Pěknicová, Jana
Seminal plasma proteins bind to the sperm surface at ejaculation and may modulate sperm properties during reproduction. Porcine spermadhesins (AQN, AWN, PSP) are secreted mainly by the seminal vesicles (SV), but their mRNAs have been found also in the cauda epididymis and prostate. Using specific polyclonal antibodies, PSP-I and PSP-II proteins were immunodetected in tissue extracts from cauda epididymis, prostate, SV and Cowper´s glands on the blots, and in secretory tissues of cauda epididymis, prostate and SV by indirect immunofluorescence. We localized PSP spermadhesins on epididymal and ejaculated spermatozoa. PSP proteins are produced not only by SV and prostate, but also by epididymis. Characterization of seminal plasma proteins expressed in the individual reproductive organs might help to understand their role in the reproduction process.
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Role of sperm surface proteins in reproduction
Jonáková, Věra ; Postlerová, Pavla ; Tichá, M. ; Pěknicová, Jana
Seminal plasma proteins bind to the sperm surface at ejaculation and may modulate sperm properties during reproduction. DQH sperm surface protein shows affinity to oviductal epithelium and zona pellucida (ZP) glycoproteins. The mRNA transcript of DQH protein was found in seminal vesicles (SV). DQH was immunodetected by monoclonal antibodies (MAbs) in SV extract and fluid, on SV tissue sections and on the membrane-associated acrosome part of ejaculated spermatozoa. These results confirmed the ability of DQH protein to bind to the sperm surface at ejaculation and to participate in the formation of the sperm reservoir in the porcine oviduct. Moreover, monoclonal antibodies reduced binding of sperm to oocytes and proved the role of DQH protein in the sperm-ZP primary binding.
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