Number of found documents: 47
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One more drop for decreasing reproduction
Dvořáková-Hortová, K.; Šídlová, A.; Děd, Lukáš; Hladovcová, D.; Vieweg, M.; Weidner, W.; Steger, K.; Stopka, P.; Paradowska-Dogan, A.
2014 - English
Toxoplasma gondii is a common protozoan parasite that infects warm-blooded animals throughout the world, including mice and humans. During infection, both, the parasite and the host, utilize various mechanisms to maximize their own reproductive success. Mice and humans are both the intermediate hosts for Toxoplasma gondii, which forms specialized vacuoles containing reproductive cysts in the formers’ tissue. As half of the human population is infected, developing a disease called toxoplasmosis, along with an ever-growing number of couples suffering with idiopathic infertility, it is therefore surprising that there is a lack of research on how T.gondii can alter reproductive parameters. In this study, a detailed histometric screening of the testicular function along with the levels of the pituitary luteinizing hormone (LH) were analysed in infected mice. Data on relative testis and epididymis weight, and sperm count were also collected. Based on the results obtained, the level of LH in the urine of Toxoplasma gondii infected mice was lower compared to the control. In direct correlation with the hormone level, testicular function and sperm production was also significantly lower in T. gondii positive group using sperm count and histometric analysis as a marker. Not only were the number of leptotene primary spermatocytes and spermatids lowered, but the number of Sertoli cells and the tubule diameter were elevated. In parallel, a pilot epigenetic study on global testicular methylation, and specific methylation of Crem, Creb1 and Hspa1genes essential for successfully ongoing spermatogenesis was performed. Global methylation was elevated in Toxoplasma infected mice, and differences in the DNA methylation of selected genes were detected between the Toxoplasma positive and control group. These findings demonstrate a direct relation between T. gondii infection and the decrease of male reproductive fitness in mice, which may contribute to an increase of infertility in humans. Keywords: Toxoplasma gondii; luteinizing hormone; spermatocytes; methylation Available at various institutes of the ASCR
One more drop for decreasing reproduction

Toxoplasma gondii is a common protozoan parasite that infects warm-blooded animals throughout the world, including mice and humans. During infection, both, the parasite and the host, utilize various ...

Dvořáková-Hortová, K.; Šídlová, A.; Děd, Lukáš; Hladovcová, D.; Vieweg, M.; Weidner, W.; Steger, K.; Stopka, P.; Paradowska-Dogan, A.
Biotechnologický ústav, 2014

Enzymatic and inhibiting activity in boar epididymal fluid
Davidová, Nina; Ren, Š.; Liberda, J.; Jonáková, Věra; Maňásková-Postlerová, Pavla
2014 - English
Sperm maturation, represents a key step in the reproduction process. Spermatozoa, particularly the plasma membrane, are exposed to epididymal fluid (EF) components representing the natural environment essential for their post-testicular maturation. Changes in the sperm membrane proteins are influenced by proteolytic and glycosidic enzymes present in the EF. Accordingly, the occurrence of inhibitors in this reproductive organ is very important for the regulation of sperm membrane protein processing. In present study, we monitored protease and glycosidase activities, and inhibitors of metallo- and serine proteinases in boar EF. Additionally, we studied acrosin inhibitor in fluid, spermatozoa and tissue along the epididymis. We chromatographically separated boar EF into several fractions. These fractions were subjected to SDS-electrophoresis and the separated proteins were either studied by zymographic methods or transferred to nitrocellulose membranes for detection of metallo- and serine proteinases and their inhibitors, and acrosin inhibitor by specific antibody, respectively. Acrosin inhibitor was monitored also in the sperm and tissue of the boar epididymis. In boar epididymal fluid, several metallo- and serine proteinases with different molecular masses, and inhibitors of metalloproteinase MMP-9 and acrosin were found. We measured strong activity of mannosidase in this fluid. Using specific antibody, we registered the increasing signal of acrosin inhibitor from caput to cauda epididymis in the spermatozoa, fluid and also tissue. Proteinases and their inhibitors in reproductive fluids may play a significant role in reproduction processes. Especially, acrosin inhibitor in the reproductive tract inactivates prematurely released sperm acrosin and protects spermatozoa and reproductive epithelium against proteolytic degradation. High mannosidase activity in boar EF suggests evident role of mannose structures in the sperm interaction during reproductive events. Keywords: enzymatic activity; inhibiting activity; epididymal fluid; proteinase Available at various institutes of the ASCR
Enzymatic and inhibiting activity in boar epididymal fluid

Sperm maturation, represents a key step in the reproduction process. Spermatozoa, particularly the plasma membrane, are exposed to epididymal fluid (EF) components representing the natural environment ...

Davidová, Nina; Ren, Š.; Liberda, J.; Jonáková, Věra; Maňásková-Postlerová, Pavla
Biotechnologický ústav, 2014

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
2014 - English
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. Keywords: monoclonal antibody; flow cytometry; spermatozoa; asthenozoospermia; bílkoviny; muži Available at various institutes of the ASCR
Flow cytometry (FCM) sperm assessment In normozoospermic and asthenozoospermic men using monoclonal antibodies against sperm proteins

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 ...

Čapková, Jana; Kubátová, Alena; Děd, Lukáš; Teplá, O.; Pěknicová, Jana
Biotechnologický ústav, 2014

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.
2014 - English
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. Keywords: tetrabromobisphenol A; protamine; spermatozoa Available at various institutes of the ASCR
The effect of tetrabromobisphenol a on protamination and DNA quality of mouse sperm

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 ...

Žatecká, Eva; Castillo, J.; Elzeinová, Fatima; Kubátová, Alena; Děd, Lukáš; Pěknicová, Jana; Oliva, R.
Biotechnologický ústav, 2014

Panel of monoclonal antibodies – alternative tool For monitoring of sperm–zona pellucida receptors localization and identification
Zigo, Michal; Dorosh, Andriy; Pohlová, Alžběta; Jonáková, Věra; Šulc, Miroslav; Maňásková-Postlerová, Pavla
2014 - English
Primary binding of the sperm to the zona pellucida (ZP) is one of the many steps necessary for successful fertilization in all sexually reproducing species. Sperm bind ZP by means of membrane receptors which recognize carbohydrate moieties on ZP glycoproteins according to a well-precised sequential process. Primary-binding receptors are localized throughout the acrosomal region of the sperm surface of which many have been disclosed in various mammals. For the monitoring sperm-zona pellucida receptors in terms of localization and characterization - panel of monoclonal antibodies against proteins from the sperm surface was prepared. Antibodies were screened by immunofluorescence and Western blotting for protein localizations and competence of antibodies, respectively. Antibodies recognizing proteins localized on the sperm head and simultaneously detected by Western blot were further studied by means of immunolocalization in reproductive tissues and fluids, binding to ZP, immunoprecipitation and protein identification using MS analysis. Out of 17 prepared antibodies, 8 antibodies were simultaneously recognizing proteins localized on the sperm head and detecting proteins of interest by Western blotting. Further only 3 antibodies recognized proteins which also coincided in binding to ZP. These 3 antibodies were used for immunoprecipitation, and further protein identification of immunoprecipitates revealed that the antibodies distinguish acrosin precursor, RAB2A protein, and lactadherin P47. Acrosin and lactadherin P47 have been already detected on the sperm surface and their physiological functions in reproduction have been proposed. To our knowledge, this is the first time RAB2A has been found on the surface of sperm and its physiological function in the process of fertilization remains undisclosed. Keywords: monoclonal antibody; zona pellucida; acrosin precursor; RAB2A; lactahedrin P47 Available at various institutes of the ASCR
Panel of monoclonal antibodies – alternative tool For monitoring of sperm–zona pellucida receptors localization and identification

Primary binding of the sperm to the zona pellucida (ZP) is one of the many steps necessary for successful fertilization in all sexually reproducing species. Sperm bind ZP by means of membrane ...

Zigo, Michal; Dorosh, Andriy; Pohlová, Alžběta; Jonáková, Věra; Šulc, Miroslav; Maňásková-Postlerová, Pavla
Biotechnologický ústav, 2014

Effect of diabetes mellitus on reproductive parameters in mice
Margaryan, Hasmik; Elzeinová, Fatima; Kubátová, Alena; Strolená, Eva; Pěknicová, Jana
2014 - English
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. Keywords: diabetes mellitus; monoclonal antibody; reproduction Available at various institutes of the ASCR
Effect of diabetes mellitus on reproductive parameters in mice

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 ...

Margaryan, Hasmik; Elzeinová, Fatima; Kubátová, Alena; Strolená, Eva; Pěknicová, Jana
Biotechnologický ústav, 2014

Changes in the expression of selected testicular genes in mice
Valášková, Eliška; Margaryan, Hasmik; Žatecká, Eva; Pěknicová, Jana
2014 - English
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. Keywords: diabetes mellitus; gene expression; reproduction Available at various institutes of the ASCR
Changes in the expression of selected testicular genes in mice

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 ...

Valášková, Eliška; Margaryan, Hasmik; Žatecká, Eva; Pěknicová, Jana
Biotechnologický ústav, 2014

The ubiquitin–proteasome system is involved in the regulation of activity of spermadhesin aqn1 and acrosin inhibitor, the two sperm surface proteins, during porcine fertilization
Jonáková, Věra; Yi, Y.J.; Postlerová, Pavla; Pěknicová, Jana
2014 - English
The spermadhesin AQN1and acrosin inhibitor (AI/SPINK2) proteins bind to the sperm plasma membrane at ejaculation. The AQN1 has been implicated in sperm binding to zona pellucida (ZP) of the oocyte as well as in sperm interactions with the epithelium of the oviductal sperm reservoir. The SPINK2 protects spermatozoa from proteolytic degradation during their trip up the female genital tract toward the oocyte. This study examined the role of two components of the 19S proteasome regulatory complex, the ubiquitin C-terminal hydrolase UCHL3 and PSMD8 in the AQN1-mediated boar sperm binding to zona pellucida. Interaction of PSMD4 subunit with the acrosomal surface-associated acrosin inhibitor AI/SPINK2 provided another line of evidence for the presence of 26S proteasomes on the sperm surface. Detection of the ubiquitinated forms of SPINK2 supports the hypothesis that SPINK2 activity is controlled by ubiquitin-proteasome system (UPS). The activity of the porcine AQN1, and thus the efficiency of sperm-oocyte recognition/binding, may be controlled by elements of the sperm surface-bound UPS, in particular by UCHL3, and by proteasomal regulatory complex subunit PSMD8. Ubiquitinated isoforms of AQN1 were also detected in boar sperm extracts. The UCHL inhibitor ubiquitin aldehyde and the antibodies against UCHL3 or PSMD8 increased the rate of sperm-ZP penetration and polyspermy during porcine in vitro fertilization (IVF). In contrast, the addition of recombinant UCHL3 to fertilization medium significantly reduced polyspermy rates, while maintaining satisfactory rate of monospermic fertilization (~50%). These results are significant for production agriculture. The high level of polyspermy that hinders porcine IVF for commercial embryo transfer could be mitigated by the modulation of the UCHL3 and/or PSMD8 activity. Keywords: ubiquitin-proteasome system; acrosin inhibitor; spermadhesin AQN1 Available at various institutes of the ASCR
The ubiquitin–proteasome system is involved in the regulation of activity of spermadhesin aqn1 and acrosin inhibitor, the two sperm surface proteins, during porcine fertilization

The spermadhesin AQN1and acrosin inhibitor (AI/SPINK2) proteins bind to the sperm plasma membrane at ejaculation. The AQN1 has been implicated in sperm binding to zona pellucida (ZP) of the oocyte as ...

Jonáková, Věra; Yi, Y.J.; Postlerová, Pavla; Pěknicová, Jana
Biotechnologický ústav, 2014

Endocrine disruptors induce transgenerational alterations of the male reproductive parameters and mirna expression profiles in mouse primordial germ cells
Děd, Lukáš; Brieno-Enríquez, M.A.; García-López, J.; Cárdenas, D.B.; Guibert, S.; Hourcade, J.de D.; Pěknicová, Jana; Weber, M.; del Mazo, J.
2014 - English
Primordial germ cells (PGCs) are the embryonic precursors of the germ cell linage, which are restricted to form only sperm and oocytes following their specification from pluripotent cells. PGC precursors are specified in the epiblast around 6.25 days post coitum (dpc), and around 7.25 dpc become identifiable in a 40 cell-cluster. In the present study, we used a mouse model to evaluate the trans-generational (F1-F3) effects of vinclozolin (VCZ) administrated in two doses on male reproductive parameters. We observed decreased fertility rate, higher apoptotic rate and histopathologic alterations in adult testis, PGC number reduction, increments of PGCs apoptosis and changes in PGCs gene expression among all three generations. In the attempt to clarify the trans-generational transmition of the altered phenotypes, we performed the microRNA expression and DNA methylation analysis. We observed the significant alteration in the expression of multiple microRNA and microRNA-regulated genes which are important for PGCs specification, including LIN28, let-7 and BLIMP1. Trans-generational deregulation in the expression of factors involved in the Lin28-let-7-Blimp1 pathway can lead to specific VCZ-induced phenotype observed in our study. Keywords: endocrine disruptor; miRNA expression; primordial germ cells; vinclozolin; DNA methylation Available at various institutes of the ASCR
Endocrine disruptors induce transgenerational alterations of the male reproductive parameters and mirna expression profiles in mouse primordial germ cells

Primordial germ cells (PGCs) are the embryonic precursors of the germ cell linage, which are restricted to form only sperm and oocytes following their specification from pluripotent cells. PGC ...

Děd, Lukáš; Brieno-Enríquez, M.A.; García-López, J.; Cárdenas, D.B.; Guibert, S.; Hourcade, J.de D.; Pěknicová, Jana; Weber, M.; del Mazo, J.
Biotechnologický ústav, 2014

Book of abstracts XXth Symposium of Biology and Immunology of Reproduction with international participation
Pěknicová, Jana; Kubátová, Alena; Elzeinová, Fatima
2014 - English
Book of abstracts. The Symposium was focused on immunology of reproduction and specific problems in reproduction (mainly in human infertility). Keywords: reproduction; immunoglobulins; monoclonal antibody; endocrine disruptors; oocytes; infection Available at various institutes of the ASCR
Book of abstracts XXth Symposium of Biology and Immunology of Reproduction with international participation

Book of abstracts. The Symposium was focused on immunology of reproduction and specific problems in reproduction (mainly in human infertility).

Pěknicová, Jana; Kubátová, Alena; Elzeinová, Fatima
Biotechnologický ústav, 2014

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