Di-(2-ethylhexyl)-phthalate (DEHP) is a compound widely used as a plasticizer, which can leach from plastics into the environment and thus influence human health. The aim of this study was to analyze whether exposure to an environmentally relevant dose of DEHP during mice fetal development or puberty can cause long-lasting changes detectable month/s after the last exposure. We used a DEHP concentration relevant to a daily human intake of 2.4-3 μg/kg of body weight/day. CD1 outbred mice were treated either in utero or postnatally during puberty and analyzed in adulthood. Analyzing fertility parameters using morphometric, histologic, genomic and proteomic methods we showed that DEHP exposure leads to decreased sperm concentration and quality, in both experimental groups. Moreover, the changes in anogenital distance, seminal vesicle weight, and testicular gene expression suggest a disturbance of androgen signaling in exposed animals. In conclusion, we hereby present, that the prenatal and pubertal exposure to a low dose of DEHP negatively influenced reproductive endpoints in male mice, and some of the effects were persistent until adulthood.
- MeSH
- anální kanál anatomie a histologie účinky léků MeSH
- diethylhexylftalát toxicita MeSH
- endokrinní disruptory toxicita MeSH
- maternofetální výměna látek MeSH
- mužské pohlavní orgány anatomie a histologie účinky léků MeSH
- myši inbrední ICR MeSH
- pohlavní dospělost účinky léků MeSH
- spermie účinky léků MeSH
- těhotenství MeSH
- testis anatomie a histologie účinky léků MeSH
- vývojová regulace genové exprese účinky léků MeSH
- zpožděný efekt prenatální expozice chemicky indukované genetika MeSH
- zvířata MeSH
- zvláčňovadla toxicita MeSH
- Check Tag
- mužské pohlaví MeSH
- těhotenství MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Capacitation involves physiological changes that spermatozoa must undergo in the female reproductive tract or in vitro to obtain the ability to bind, penetrate and fertilize the egg. Up to date, several methods have been developed to characterize this complex biological process. The goal of the presented study is to mutually compare several fluorescent techniques, check their ability to detect changes in molecular processes during the capacitation progress and determine their ability to predict the percentage of acrosome reacted (AR) sperm after the exposure to solubilized zona pellucida (ZP). The capacitation process was analyzed using four fluorescent techniques: 1. chlortetracycline (CTC) staining, 2. anti-acrosin antibody (ACR.2) assay, 3. anti-phosphotyrosine (pY) antibody assay, 4. fluorescein isothiocyanate-conjugated phalloidin (FITC-phall) assay. All these methods were tested using fluorescent microscopy and flow cytometry. RESULTS: All selected methods are capable to detect the capacitation progress of boar sperm in vitro, but there are significant differences in their outcome when using fluorescent microscopy or flow cytometry experimental arrangements and subsequent statistical analysis (KW-ANOVA). Also, the ability to predict the absolute numbers of sperm which will undergo ZP-induced AR differ significantly (CTC and ACR.2 gave the best predictions). CONCLUSIONS: Our study compared four largely used methods used to characterize capacitation process, highlighted their differences and showed that all are able to detect capacitation progress, CTC and ACR.2 are furthermore able to accurately predict the percentage of AR sperm after ZP-induced AR.
- MeSH
- akrozomální reakce fyziologie MeSH
- faloidin MeSH
- fluorescein-5-isothiokyanát MeSH
- fluorescenční barviva * MeSH
- fluorescenční mikroskopie * metody MeSH
- fluorescenční protilátková technika MeSH
- kapacitace spermií fyziologie MeSH
- průtoková cytometrie * metody MeSH
- spermie fyziologie MeSH
- Sus scrofa fyziologie MeSH
- vápník analýza MeSH
- zona pellucida fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Pro detekci molekulárních změn vyvolaných diabetes mellitus jsme použili panel protilátek proti vybraným proteinům testes a spermií. Detegovali jsme změny v morfologii a histologii varlat a změny v expresi důležitých proteinů, které jsou zapojeny do spermatogeneze a mezibuněčné komunikace. Monoklonální protilátky proti intra -akrozomálním proteinům prokázaly zhoršenou kvalitu spermií u diabetických pacientů i spermií diabetických myší. Panel vybraných protilátek lze použít k testování změn v testes a kvalitě spermií vyvolaných diabetem. Tyto výsledky jasně dokumentují vliv diabetického prostředí na morfologii a histologii varlat a kvalitu spermií.
We used a panel of antibodies against selected testicular and sperm proteins to detect molecular changes induced by diabetes mellitus. We detected changes in morphology and histology of the testes, and changes in the expression and distribution of proteins involved in spermatogenesis and intercellular communication. Monoclonal antibodies against intra -acrosomal proteins demonstrated the impaired quality of sperms from diabetic patients as well as sperms from diabetic mice. In conclusion, the panel of selected antibodies can be used to test diabetes -induced changes in the testes and sperm quality. These results clearly document the effect of the diabetic environment on the morphology and histology of the testes, and sperm quality.
The crucial role that oestrogens play in male reproduction has been generally accepted; however, the exact mechanism of their action is not entirely clear and there is still much more to be clarified. The oestrogen response is mediated through oestrogen receptors, as well as classical oestrogen receptors' variants, and their specific co-expression plays a critical role. The importance of oestrogen signalling in male fertility is indicated by the adverse effects of selected oestrogen-like compounds, and their interaction with oestrogen receptors was proven to cause pathologies. The aims of this review are to summarise the current knowledge on oestrogen signalling during spermatogenesis and sperm maturation and discuss the available information on oestrogen receptors and their splice variants. An overview is given of species-specific differences including in humans, along with a detailed summary of the methodology outcome, including all the genetically manipulated models available to date. This review provides coherent information on the recently discovered mechanisms of oestrogens' and oestrogen receptors' effects and action in both testicular somatic and germ cells, as well as in mature sperm, available for mammals, including humans.
- MeSH
- aromatasa nedostatek genetika MeSH
- estrogeny farmakologie MeSH
- lidé MeSH
- receptory pro estrogeny metabolismus MeSH
- signální transdukce MeSH
- spermatogeneze účinky léků MeSH
- testis účinky léků metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Estrogens play a crucial role in spermatogenesis and estrogen receptor α knock-out male mice are infertile. It has been demonstrated that estrogens significantly increase the speed of capacitation in vitro; however this may lead to the reduction of reproductive potential due to the decreased ability of these sperm to undergo the acrosome reaction. To date the in vivo effect of estrogens on the ability of sperm to capacitate has not been investigated. Therefore, in this study, we exposed mice (n=24) to 17β-estradiol (E2) at the concentration of 20 ng/ml either during puberty from the fourth to seventh week of age (n=8), or continuously from birth for a period of 12 weeks (n=8) at which age the animals from both groups were killed. The capacitation status of epididymal and testicular sperm was analysed by tyrosine phosphorylation (TyrP) antibody (immunofluorescence and western blot) and chlortetracycline (CTC) assay. According to our results, in vivo exposure to increased E2 concentrations caused premature sperm capacitation in the epididymis. The effect of E2, however, seems reversible because after the termination of the exposure premature epididymal sperm capacitation is decreased in animals treated during puberty. Furthermore the changes in epididymal sperm capacitation status detected by TyrP and CTC positively correlate with plasma levels of E2 and the expression of the estrogen-dependent trefoil factor 1 (Tff1) gene in testicular tissue. Therefore, our data implicate that in vivo exposure to E2 under specific conditions leads to the premature capacitation of mouse sperm in epididymis with a potential negative impact on the sperm reproductive fitness in the female reproductive tract.
- MeSH
- biologické markery metabolismus MeSH
- časové faktory MeSH
- chlortetracyklin metabolismus MeSH
- epididymis účinky léků metabolismus patologie MeSH
- estradiol aplikace a dávkování krev MeSH
- fluorescenční protilátková technika MeSH
- fosforylace MeSH
- kapacitace spermií účinky léků MeSH
- myši MeSH
- peptidy genetika metabolismus MeSH
- rozvrh dávkování léků MeSH
- sexuální vývoj MeSH
- spermie účinky léků metabolismus patologie MeSH
- testis účinky léků metabolismus patologie MeSH
- tyrosin MeSH
- věkové faktory MeSH
- western blotting MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Tetrabromobisphenol A (TBBPA) is a substance widely used in industry as a flame retardant. TBBPA was found in the environment and was detected even in the human body. The effect of this chemical was observed in different cell lines in vitro and it is supposed that TBBPA may affect various hormonal systems in vivo. In this study we examined the effect of TBBPA on the reproductive parameters of two generations of outbred mice in vivo. Experimental and control animals of F1 generation were bred in various conditions to enable evaluation of the possible trans-generational effect. An increased incidence of apoptosis in the testes and changes in the morphometry of seminiferous tubules was detected in the experimental animals. In addition, changes in the expression pattern of selected genes encoding proteins that play an important role during spermatogenesis were observed. In contrast, sperm quality and reproduction were not affected by TBBPA.
- MeSH
- apoptóza účinky léků MeSH
- myši MeSH
- polybrombifenylové sloučeniny toxicita MeSH
- regulace genové exprese účinky léků MeSH
- retardanty hoření toxicita MeSH
- spermie účinky léků MeSH
- testis anatomie a histologie účinky léků metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Mammalian sperm must undergo a series of controlled molecular processes in the female reproductive tract called capacitation before they are capable of penetrating and fertilizing the egg. Capacitation, as a complex biological process, is influenced by many molecular factors, among which steroidal hormone estrogens play their role. Estrogens, present in a high concentration in the female reproductive tract are generally considered as primarily female hormones. However, there is increasing evidence of their important impact on male reproductive parameters. The purpose of this study is to investigate the effect of three natural estrogens such as estrone (E1), 17beta-estradiol (E2) and estriol (E3) as well as the synthetical one, 17 alpha-ethynylestradiol (EE2) on boar sperm capacitation in vitro. METHODS: Boar sperm were capacitated in vitro in presence of estrogens. Capacitation progress in control and experimental samples was analyzed by flow cytometry with the anti-acrosin monoclonal antibody (ACR.2) at selected times of incubation. Sperm samples were analyzed at 120 min of capacitation by CTC (chlortetracycline) assay, immunocytochemistry and flow cytometry with anti-acrosin ACR.2 antibody. Furthermore, sperm samples and capacitating media were analyzed by immunocytochemistry, ELISA with the ACR.2 antibody, and the acrosin activity assay after induced acrosomal reaction (AR). RESULTS: Estrogens stimulate sperm capacitation of boar sperm collected from different individuals. The stimulatory effect depends on capacitation time and is highly influenced by differences in the response to estrogens such as E2 by individual animals. Individual estrogens have relatively same effect on capacitation progress. In the boar samples with high estrogen responsiveness, estrogens stimulate the capacitation progress in a concentration-dependent manner. Furthermore, estrogens significantly increase the number of acrosome-reacted sperm after zona pellucida- induced acrosomal reaction. CONCLUSIONS: We demonstrate here the stimulatory effect of four different estrogens on boar sperm capacitation in vitro. According to our results, there is significant difference in the response to tested estrogens at different capacitation time and among individual animals. In animals with a high response to estrogens, there is a concentration-dependent stimulation of capacitation and individual estrogens have relatively the same effect. Effects of individual estrogens, differences in the response to them by individual animals, their time and concentration-dependent outcomes further contribute to our knowledge about steroidal action in sperm.
- MeSH
- časové faktory MeSH
- estradiol farmakologie MeSH
- estrogeny farmakologie MeSH
- fluorescence MeSH
- kapacitace spermií účinky léků fyziologie MeSH
- kultivované buňky MeSH
- počet spermií MeSH
- průtoková cytometrie MeSH
- Sus scrofa fyziologie MeSH
- zona pellucida účinky léků metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
