Limits...
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.

Show MeSH

Related in: MedlinePlus

Differences in capacitation and AR progress between samples A-D exposed by 1 μM E2 and control samples measured by flow cytometry with ACR.2 antibody. Representative pictures of FITC channel histograms at 0, 60, 120, 180, 240 min and after induced AR. Control samples in black, experimental samples in green. The increase in fluorescent intensity (right peak) corresponds to the capacitation progress. Differences among the control and experimental samples in arithmetic mean of the fluorescent intensity in the FITC channel were assessed by t-test. *P < 0.05, **P < 0.01, ***P < 0.001. The most significant difference is recognizable at 120 min.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC2908632&req=5

Figure 3: Differences in capacitation and AR progress between samples A-D exposed by 1 μM E2 and control samples measured by flow cytometry with ACR.2 antibody. Representative pictures of FITC channel histograms at 0, 60, 120, 180, 240 min and after induced AR. Control samples in black, experimental samples in green. The increase in fluorescent intensity (right peak) corresponds to the capacitation progress. Differences among the control and experimental samples in arithmetic mean of the fluorescent intensity in the FITC channel were assessed by t-test. *P < 0.05, **P < 0.01, ***P < 0.001. The most significant difference is recognizable at 120 min.

Mentions: In order to determine the potential differences in the capacitation progress between control and experimental groups, sperm samples were analyzed by flow cytometry with ACR.2 antibody at selected times of capacitation. The experimental sample was capacitated with E2 at a 1 μM concentration, and the control sample with the same amount of ethanol as in the experimental sample. 1 μM concentration of E2 was selected based on the previous mouse study [18] where it was defined as the lowest concentration with any significant effect on sperm capacitation. Sperm were collected at 0, 10, 30, 60, 120, 180, and 240 min of capacitation and after the induced acrosomal reaction. The first significant difference between the control and experimental group was at 60 min capacitation in the arithmetic mean of the fluorescent intensity in the FITC channel (Fig. 3). The strongest significant difference was then at 120 min of capacitation. After an induced acrosomal reaction, a significantly higher number of sperm underwent AR in the experimental group in comparison with the control one.


Effect of estrogens on boar sperm capacitation in vitro.

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

Differences in capacitation and AR progress between samples A-D exposed by 1 μM E2 and control samples measured by flow cytometry with ACR.2 antibody. Representative pictures of FITC channel histograms at 0, 60, 120, 180, 240 min and after induced AR. Control samples in black, experimental samples in green. The increase in fluorescent intensity (right peak) corresponds to the capacitation progress. Differences among the control and experimental samples in arithmetic mean of the fluorescent intensity in the FITC channel were assessed by t-test. *P < 0.05, **P < 0.01, ***P < 0.001. The most significant difference is recognizable at 120 min.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 3: Differences in capacitation and AR progress between samples A-D exposed by 1 μM E2 and control samples measured by flow cytometry with ACR.2 antibody. Representative pictures of FITC channel histograms at 0, 60, 120, 180, 240 min and after induced AR. Control samples in black, experimental samples in green. The increase in fluorescent intensity (right peak) corresponds to the capacitation progress. Differences among the control and experimental samples in arithmetic mean of the fluorescent intensity in the FITC channel were assessed by t-test. *P < 0.05, **P < 0.01, ***P < 0.001. The most significant difference is recognizable at 120 min.
Mentions: In order to determine the potential differences in the capacitation progress between control and experimental groups, sperm samples were analyzed by flow cytometry with ACR.2 antibody at selected times of capacitation. The experimental sample was capacitated with E2 at a 1 μM concentration, and the control sample with the same amount of ethanol as in the experimental sample. 1 μM concentration of E2 was selected based on the previous mouse study [18] where it was defined as the lowest concentration with any significant effect on sperm capacitation. Sperm were collected at 0, 10, 30, 60, 120, 180, and 240 min of capacitation and after the induced acrosomal reaction. The first significant difference between the control and experimental group was at 60 min capacitation in the arithmetic mean of the fluorescent intensity in the FITC channel (Fig. 3). The strongest significant difference was then at 120 min of capacitation. After an induced acrosomal reaction, a significantly higher number of sperm underwent AR in the experimental group in comparison with the control one.

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