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Effect of estrogens on boar sperm capacitation in vitro.

Ded L, Dostalova P, Dorosh A, Dvorakova-Hortova K, Peknicova J - Reprod. Biol. Endocrinol. (2010)

Bottom Line: Individual estrogens have relatively same effect on capacitation progress.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Diagnostics for Reproductive Medicine, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.

ABSTRACT

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.

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Concentration of acrosin in medium after induced AR measured enzymatically by acrosin assay and immunochemically by ELISA with ACR.2 antibody. After 240 min of the in vitro capacitation process, experimental samples from boars A-D with 1 μM concentration of four estrogens were treated by boar zona pellucida to induce AR. Capacitation media were analyzed by ELISA with ACR.2 antibody and acrosin assay to biochemically determine the number of sperm after AR. Differences were analyzed by ANOVA; post hoc comparison was performed by Newman-Keuls test. Whiskers denote ± SE. *P < 0.05, **P < 0.01, ***P < 0.001.
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Figure 7: Concentration of acrosin in medium after induced AR measured enzymatically by acrosin assay and immunochemically by ELISA with ACR.2 antibody. After 240 min of the in vitro capacitation process, experimental samples from boars A-D with 1 μM concentration of four estrogens were treated by boar zona pellucida to induce AR. Capacitation media were analyzed by ELISA with ACR.2 antibody and acrosin assay to biochemically determine the number of sperm after AR. Differences were analyzed by ANOVA; post hoc comparison was performed by Newman-Keuls test. Whiskers denote ± SE. *P < 0.05, **P < 0.01, ***P < 0.001.

Mentions: Sperm samples were analyzed after 240 min of capacitation and induced AR by CTC, immunocytochemistry and ELISA with ACR.2 antibody. The acrosin assay was used to further evaluate the effect of estrogens on capacitation and the acrosomal reaction. There was a significantly higher number of sperm, which underwent ZP -induced AR in all experimental samples in comparison with the experimental group (Fig. 6). Data from immunocytochemistry were further verified by ELISA with ACR.2 antibody and the acrosin assay (Fig. 7).


Effect of estrogens on boar sperm capacitation in vitro.

Ded L, Dostalova P, Dorosh A, Dvorakova-Hortova K, Peknicova J - Reprod. Biol. Endocrinol. (2010)

Concentration of acrosin in medium after induced AR measured enzymatically by acrosin assay and immunochemically by ELISA with ACR.2 antibody. After 240 min of the in vitro capacitation process, experimental samples from boars A-D with 1 μM concentration of four estrogens were treated by boar zona pellucida to induce AR. Capacitation media were analyzed by ELISA with ACR.2 antibody and acrosin assay to biochemically determine the number of sperm after AR. Differences were analyzed by ANOVA; post hoc comparison was performed by Newman-Keuls test. Whiskers denote ± SE. *P < 0.05, **P < 0.01, ***P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2908632&req=5

Figure 7: Concentration of acrosin in medium after induced AR measured enzymatically by acrosin assay and immunochemically by ELISA with ACR.2 antibody. After 240 min of the in vitro capacitation process, experimental samples from boars A-D with 1 μM concentration of four estrogens were treated by boar zona pellucida to induce AR. Capacitation media were analyzed by ELISA with ACR.2 antibody and acrosin assay to biochemically determine the number of sperm after AR. Differences were analyzed by ANOVA; post hoc comparison was performed by Newman-Keuls test. Whiskers denote ± SE. *P < 0.05, **P < 0.01, ***P < 0.001.
Mentions: Sperm samples were analyzed after 240 min of capacitation and induced AR by CTC, immunocytochemistry and ELISA with ACR.2 antibody. The acrosin assay was used to further evaluate the effect of estrogens on capacitation and the acrosomal reaction. There was a significantly higher number of sperm, which underwent ZP -induced AR in all experimental samples in comparison with the experimental group (Fig. 6). Data from immunocytochemistry were further verified by ELISA with ACR.2 antibody and the acrosin assay (Fig. 7).

Bottom Line: Individual estrogens have relatively same effect on capacitation progress.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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Diagnostics for Reproductive Medicine, Institute of Biotechnology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.

ABSTRACT

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.

Show MeSH
Related in: MedlinePlus