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Estrogen suppresses melatonin-enhanced hyperactivation of hamster spermatozoa.

Fujinoki M, Takei GL - J. Reprod. Dev. (2015)

Bottom Line: Although it has been indicated that melatonin also enhances hyperactivation, it is unknown whether melatonin-enhanced hyperactivation is also suppressed by 17βE2 and GABA.In the present study, melatonin-enhanced hyperactivation was significantly suppressed by 17βE2 but not by GABA.These results suggest that enhancement of hyperactivation is regulated by melatonin and 17βE2 through non-genomic regulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Dokkyo Medical University, Tochigi 321-0293, Japan.

ABSTRACT
Hamster sperm hyperactivation is enhanced by progesterone, and this progesterone-enhanced hyperactivation is suppressed by 17β-estradiol (17βE2) and γ-aminobutyric acid (GABA). Although it has been indicated that melatonin also enhances hyperactivation, it is unknown whether melatonin-enhanced hyperactivation is also suppressed by 17βE2 and GABA. In the present study, melatonin-enhanced hyperactivation was significantly suppressed by 17βE2 but not by GABA. Moreover, suppression of melatonin-enhanced hyperactivation by 17βE2 occurred through non-genomic regulation via the estrogen receptor (ER). These results suggest that enhancement of hyperactivation is regulated by melatonin and 17βE2 through non-genomic regulation.

No MeSH data available.


Related in: MedlinePlus

Inhibition of suppression of melatonin-enhanced hyperactivation by 17βE2. The percentages of motile (A, C) and hyperactivated (B, D) spermatozoa are shown (A, B) for when spermatozoa were exposed to melatonin and tamoxifen, and (C, D) for when spermatozoa were exposed to melatonin, 17βE2 and tamoxifen. In A and B, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, melatonin or vehicle was added to the same medium. In C and D, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, 17βE2 or vehicle was added to the same medium. After incubation for 5 min again, melatonin or vehicle was added to the same medium. Data are expressed as the mean ± SD. (A, B) (Control): mTALP medium with 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Tamoxifen → Melatonin): mTALP medium with addition of 1 nM melatonin, 1 μM tamoxifen, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO. (C, D) (Control): mTALP medium with 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (17βE2 → Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 0.2% (v/v) EtOH and 0.1% (v/v)DMSO; (Tamoxifen → 17βE2 →Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 1 μM tamoxifen, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO. * Significant difference compared with “Control” (P < 0.05). ** Significant difference compared with “Control” and “17βE2 → Melatonin” (P < 0.05). DMSO, dimethyl sulfoxide; 17βE2, 17β-estradiol; EtOH, ethanol; mTALP medium, modified Tyrode’s albumin lactate pyruvate medium.
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fig_003: Inhibition of suppression of melatonin-enhanced hyperactivation by 17βE2. The percentages of motile (A, C) and hyperactivated (B, D) spermatozoa are shown (A, B) for when spermatozoa were exposed to melatonin and tamoxifen, and (C, D) for when spermatozoa were exposed to melatonin, 17βE2 and tamoxifen. In A and B, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, melatonin or vehicle was added to the same medium. In C and D, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, 17βE2 or vehicle was added to the same medium. After incubation for 5 min again, melatonin or vehicle was added to the same medium. Data are expressed as the mean ± SD. (A, B) (Control): mTALP medium with 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Tamoxifen → Melatonin): mTALP medium with addition of 1 nM melatonin, 1 μM tamoxifen, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO. (C, D) (Control): mTALP medium with 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (17βE2 → Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 0.2% (v/v) EtOH and 0.1% (v/v)DMSO; (Tamoxifen → 17βE2 →Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 1 μM tamoxifen, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO. * Significant difference compared with “Control” (P < 0.05). ** Significant difference compared with “Control” and “17βE2 → Melatonin” (P < 0.05). DMSO, dimethyl sulfoxide; 17βE2, 17β-estradiol; EtOH, ethanol; mTALP medium, modified Tyrode’s albumin lactate pyruvate medium.

Mentions: Hyperactivated spermatozoa were prepared using the modified Tyrode’s albumin lactate pyruvate (mTALP) medium [20] containing 101.02 mM NaCl, 2.68 mM KCl, 2 mM CaCl2-2H2O, 1.5 mM MgCl2-6H2O, 360 μM NaH2PO4-2H2O, 35.70 mM NaHCO3, 4.5 mM D-glucose, 90 μM sodium pyruvate, 9 mM sodium lactate, 500 μM hypotaurine, 50 μM (-)epinephrine, 200 μM sodium taurocholate, 5.26 μM sodium metabisulfite, 0.05% (w/v) streptomycin sulfate, 0.05% (w/v) potassium penicillin G and 15 mg/ml BSA (pH 7.4 at 37 C under 5% (v/v)CO2 in air) according to the method described previously [19]. A drop (~ 5 μl) of posterior epididymal spermatozoa was placed in a culture dish (diameter, 35 mm) with 3 ml of the mTALP medium, followed by incubation at 37 C for 5 min to allow spermatozoa to swim up. All of the mTALP medium containing motile spermatozoa was placed in a new culture dish and incubated for 4 h at 37 C under 5% (v/v) CO2 in air to allow hyperactivation to occur. As stock solutions, melatonin (1 μM), 17αE2 (20 μg/ml) and 17βE2 (20 μg/ml) were dissolved in ethanol; tamoxifen was dissolved at 1 mM in dimethyl sulfoxide; and GABA (5 mM) and BSA-E2 (7.4 μM) were dissolved in pure water. GABA, 17αE2, 17βE2, tamoxifen or vehicle was added to the mTALP medium after swim up, and after 5 min of incubation, melatonin or vehicle was added to the mTALP medium (Figs. 1, 2, 3A, 3B, 5A, 5BFig. 1.


Estrogen suppresses melatonin-enhanced hyperactivation of hamster spermatozoa.

Fujinoki M, Takei GL - J. Reprod. Dev. (2015)

Inhibition of suppression of melatonin-enhanced hyperactivation by 17βE2. The percentages of motile (A, C) and hyperactivated (B, D) spermatozoa are shown (A, B) for when spermatozoa were exposed to melatonin and tamoxifen, and (C, D) for when spermatozoa were exposed to melatonin, 17βE2 and tamoxifen. In A and B, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, melatonin or vehicle was added to the same medium. In C and D, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, 17βE2 or vehicle was added to the same medium. After incubation for 5 min again, melatonin or vehicle was added to the same medium. Data are expressed as the mean ± SD. (A, B) (Control): mTALP medium with 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Tamoxifen → Melatonin): mTALP medium with addition of 1 nM melatonin, 1 μM tamoxifen, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO. (C, D) (Control): mTALP medium with 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (17βE2 → Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 0.2% (v/v) EtOH and 0.1% (v/v)DMSO; (Tamoxifen → 17βE2 →Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 1 μM tamoxifen, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO. * Significant difference compared with “Control” (P < 0.05). ** Significant difference compared with “Control” and “17βE2 → Melatonin” (P < 0.05). DMSO, dimethyl sulfoxide; 17βE2, 17β-estradiol; EtOH, ethanol; mTALP medium, modified Tyrode’s albumin lactate pyruvate medium.
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fig_003: Inhibition of suppression of melatonin-enhanced hyperactivation by 17βE2. The percentages of motile (A, C) and hyperactivated (B, D) spermatozoa are shown (A, B) for when spermatozoa were exposed to melatonin and tamoxifen, and (C, D) for when spermatozoa were exposed to melatonin, 17βE2 and tamoxifen. In A and B, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, melatonin or vehicle was added to the same medium. In C and D, after swim up, tamoxifen or vehicle was added to the mTALP medium containing motile spermatozoa. After incubation for 5 min, 17βE2 or vehicle was added to the same medium. After incubation for 5 min again, melatonin or vehicle was added to the same medium. Data are expressed as the mean ± SD. (A, B) (Control): mTALP medium with 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO; (Tamoxifen → Melatonin): mTALP medium with addition of 1 nM melatonin, 1 μM tamoxifen, 0.1% (v/v) EtOH and 0.1% (v/v) DMSO. (C, D) (Control): mTALP medium with 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (Melatonin): mTALP medium with addition of 1 nM melatonin, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO; (17βE2 → Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 0.2% (v/v) EtOH and 0.1% (v/v)DMSO; (Tamoxifen → 17βE2 →Melatonin): mTALP medium with addition of 1 nM melatonin, 20 ng/ml 17βE2, 1 μM tamoxifen, 0.2% (v/v) EtOH and 0.1% (v/v) DMSO. * Significant difference compared with “Control” (P < 0.05). ** Significant difference compared with “Control” and “17βE2 → Melatonin” (P < 0.05). DMSO, dimethyl sulfoxide; 17βE2, 17β-estradiol; EtOH, ethanol; mTALP medium, modified Tyrode’s albumin lactate pyruvate medium.
Mentions: Hyperactivated spermatozoa were prepared using the modified Tyrode’s albumin lactate pyruvate (mTALP) medium [20] containing 101.02 mM NaCl, 2.68 mM KCl, 2 mM CaCl2-2H2O, 1.5 mM MgCl2-6H2O, 360 μM NaH2PO4-2H2O, 35.70 mM NaHCO3, 4.5 mM D-glucose, 90 μM sodium pyruvate, 9 mM sodium lactate, 500 μM hypotaurine, 50 μM (-)epinephrine, 200 μM sodium taurocholate, 5.26 μM sodium metabisulfite, 0.05% (w/v) streptomycin sulfate, 0.05% (w/v) potassium penicillin G and 15 mg/ml BSA (pH 7.4 at 37 C under 5% (v/v)CO2 in air) according to the method described previously [19]. A drop (~ 5 μl) of posterior epididymal spermatozoa was placed in a culture dish (diameter, 35 mm) with 3 ml of the mTALP medium, followed by incubation at 37 C for 5 min to allow spermatozoa to swim up. All of the mTALP medium containing motile spermatozoa was placed in a new culture dish and incubated for 4 h at 37 C under 5% (v/v) CO2 in air to allow hyperactivation to occur. As stock solutions, melatonin (1 μM), 17αE2 (20 μg/ml) and 17βE2 (20 μg/ml) were dissolved in ethanol; tamoxifen was dissolved at 1 mM in dimethyl sulfoxide; and GABA (5 mM) and BSA-E2 (7.4 μM) were dissolved in pure water. GABA, 17αE2, 17βE2, tamoxifen or vehicle was added to the mTALP medium after swim up, and after 5 min of incubation, melatonin or vehicle was added to the mTALP medium (Figs. 1, 2, 3A, 3B, 5A, 5BFig. 1.

Bottom Line: Although it has been indicated that melatonin also enhances hyperactivation, it is unknown whether melatonin-enhanced hyperactivation is also suppressed by 17βE2 and GABA.In the present study, melatonin-enhanced hyperactivation was significantly suppressed by 17βE2 but not by GABA.These results suggest that enhancement of hyperactivation is regulated by melatonin and 17βE2 through non-genomic regulation.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Dokkyo Medical University, Tochigi 321-0293, Japan.

ABSTRACT
Hamster sperm hyperactivation is enhanced by progesterone, and this progesterone-enhanced hyperactivation is suppressed by 17β-estradiol (17βE2) and γ-aminobutyric acid (GABA). Although it has been indicated that melatonin also enhances hyperactivation, it is unknown whether melatonin-enhanced hyperactivation is also suppressed by 17βE2 and GABA. In the present study, melatonin-enhanced hyperactivation was significantly suppressed by 17βE2 but not by GABA. Moreover, suppression of melatonin-enhanced hyperactivation by 17βE2 occurred through non-genomic regulation via the estrogen receptor (ER). These results suggest that enhancement of hyperactivation is regulated by melatonin and 17βE2 through non-genomic regulation.

No MeSH data available.


Related in: MedlinePlus