• The Korean Journal of Physiology and Pharmacology
• /
• v.23 no.5
• /
• pp.381-392
• /
• 2019

Sperm function and male fertility are closely related to pH dependent $K^+$ current (KSper) in human sperm, which is most likely composed of Slo3 and its auxiliary subunit leucine-rich repeat-containing protein 52 (LRRC52). Onion peel extract (OPE) and its major active ingredient quercetin are widely used as fertility enhancers; however, the effect of OPE and quercetin on Slo3 has not been elucidated. The purpose of this study is to investigate the effect of quercetin on human Slo3 channels. Human Slo3 and LRRC52 were co-transfected into HEK293 cells and pharmacological properties were studied with the whole cell patch clamp technique. We successfully expressed and measured pH sensitive and calcium insensitive Slo3 currents in HEK293 cells. We found that OPE and its key ingredient quercetin inhibit Slo3 currents. Inhibition by quercetin is dose dependent and this degree of inhibition decreases with elevating internal alkalization and internal free calcium concentrations. Functional moieties in the quercetin polyphenolic ring govern the degree of inhibition of Slo3 by quercetin, and the composition of such functional moieties are sensitive to the pH of the medium. These results suggest that quercetin inhibits Slo3 in a pH and calcium dependent manner. Therefore, we surmise that quercetin induced depolarization in spermatozoa may enhance the voltage gated proton channel (Hv1), and activate non-selective cation channels of sperm (CatSper) dependent calcium influx to trigger sperm capacitation and acrosome reaction.

• Clinical and Experimental Reproductive Medicine
• /
• v.49 no.4
• /
• pp.277-284
• /
• 2022

Objective: Oxidative stress is a key player in the development of idiopathic male infertility (IMI), and various antioxidants have been used for the treatment of IMI with inconsistent results. Coenzyme Q10 (CoQ10) is a cofactor and an antioxidant that may improve semen parameters and reduce oxidative stress in patients with idiopathic oligoasthenospermia (OA). Therefore, this study aimed to explore the effect of CoQ10 on semen parameters and antioxidant markers in patients with idiopathic OA. Methods: Fifty patients with idiopathic OA and 35 fertile controls were enrolled in this prospective controlled study. All participants underwent a comprehensive fertility assessment. All patients received CoQ10 (300 mg/day) orally once daily for 3 months. Semen parameters, seminal CoQ10 levels, reactive oxygen species (ROS) levels, total antioxidant capacity (TAC), superoxide dismutase (SOD), and glutathione peroxidase (GPx) were measured in patients and controls at the start of the study and after 3 months. Results: Treatment with CoQ10 resulted in increased sperm progressive motility (p<0.05), total motility (p<0.01), seminal TAC (p<0.01), SOD (p<0.05), GPx (p<0.001), and seminal CoQ10 (p<0.001) levels and reduced ROS (p<0.01) in patients as compared to baseline. Sperm concentration and motility were also significantly correlated with antioxidant measures and seminal CoQ10 levels (r=0.38-0.57). Conclusion: CoQ10 therapy (300 mg/day for 3 months) improved sperm motility and seminal antioxidant markers in patients with idiopathic OA. Therefore, CoQ10 could be a promising treatment for patients with idiopathic infertility and may improve their fertility potential.

• Reproductive and Developmental Biology
• /
• v.39 no.4
• /
• pp.117-125
• /
• 2015

Iron is required for cell viability but is toxic in excess. While the iron-mediated malfunction of testicular cells is well appreciated, the underlying mechanism(s) of this effect and its relationship with fertility are poorly understood. Ferritin is a ubiquitous intracellular protein that controls iron storage, ferroxidase activity, immune response, and stress response in cells. Ferritin light chain protein (FTL) is the light subunit of the Ferritin. Previously, we had identified the FTL in bovine spermatozoa following capacitation. In present study, to investigate the role of Ferritin in sperm function, mice spermatozoa were incubated with multiple doses (1, 10 and $100{\mu}M$) of sodium nitroprusside (SNP), an iron donor. SNP was increased Ferritin levels in a dose-dependent manner. The Ferritin was detected on the acrosome in spermatozoa by immunocytochemistry. Short-term exposure of spermatozoa to SNP increased tyrosine phosphorylation and the acrosome reaction (AR). Finally, SNP affected a significant decrease in the rate of fertilization as well as blastocyst formation during early embryonic development. On the basis of these results, we propose that the effects of Ferritin on the AR may reduce overall sperm function leads to poor fertility in males and compromised embryonic development.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2003.11a
• /
• pp.107-108
• /
• 2003

This study was conducted to investigate the effects of dilution rate and stored temperature of semen at 5, 25 and 35$^{\circ}C$ on fertility in liquid rooster semen. At 5$^{\circ}C$ cold temperature, no significant difference were found in sperm mobilities on dilution rate(1:1, 1:3, 1.6) among treatments. Sperm mobility for the conservation of 3 hours at 25∼35$^{\circ}C$ were significantly higher for 1:3 and 1:6 dilution rate(semen：diluent) groups than for 1:1 dilution rate group(P<0.05). In Fertility results after artificial insemination with the conservation of 3 hours at 5∼25$^{\circ}C$ temperature, no significant difference were found in fertility on dilution rate among treatments. Fertilities after artificial insemination with the conservation of 3 hours at 35$^{\circ}C$ were significantly higher for 1.3 and 1:6 dilution rate(semen：diluent) groups than for 1：1 dilution rate group(P<0.05).

• Clinical and Experimental Reproductive Medicine
• /
• v.48 no.4
• /
• pp.352-361
• /
• 2021

Objective: The study assessed the developmental potential of germinal vesicle (GV) oocytes subjected to in vitro maturation (IVM) after prematuration culture with cilostamide (a phosphodiesterase-3 inhibitor) and the impact of cilostamide exposure on the morphology of meiosis II (MII) oocytes and subsequent embryo quality. Methods: In total, 994 oocytes were collected from 63 patients. Among 307 GV oocytes, 140 oocytes were selected for the experimental group and 130 oocytes for the control group. The denuded GV-stage oocytes were cultured for 6 hours with cilostamide in the experimental group and without cilostamide in the control group. After 6 hours, the oocytes in the experimental group were washed and transferred to fresh IVM medium. The maturational status of the oocytes in both groups was examined at 26, 36, and 48 hours. Fertilization was assessed at 18 hours post-intracytoplasmic sperm injection. Embryo quality was assessed on days 3 and 5. Results: In total, 92.1% of the oocytes remained in the GV stage, while 6.4% converted to the MI stage (p<0.01) after cilostamide exposure. In both groups, more MII oocytes were observed at 36 hours (25.8% vs. 21.5%) than at 26 hours (10.8% vs. 14.6%) and 48 hours (13% vs. 7.9%) (p>0.05). With the advent of cilostamide, blastocyst quality was better in the experimental group than in the control group (p<0.05). Conclusion: Cilostamide effectively blocked nuclear maturation and promoted cytoplasmic growth. Prematuration culture with cilostamide enabled synchronization between cytoplasmic and nuclear maturity, resulting in better blastocyst outcomes.

• Clinical and Experimental Reproductive Medicine
• /
• v.48 no.3
• /
• pp.255-261
• /
• 2021

Objective: This study aimed to investigate sperm motility and its changes after preparation as predictors of pregnancy in intrauterine insemination (IUI) cycles. Methods: In total, 297 IUI cycles from January 2012 to December 2017 at a single tertiary hospital were retrospectively analyzed. Patient and cycle characteristics, and sperm motility characteristics before and after processing were compared according to clinical pregnancy or live birth as outcomes. Results: The overall clinical pregnancy rate per cycle was 14.5% (43/297) and the live birth rate was 10.4% (30/289). Patient and cycle characteristics were similar between pregnant and non-pregnant groups. Sperm motility after preparation and the total motile sperm count before and after processing were comparable in terms of pregnancy outcomes. Pre-preparation sperm motility was significantly higher in groups with clinical pregnancy and live birth than in cycles not resulting in pregnancy (71.4%±10.9% vs. 67.2%±11.7%, p=0.020 and 71.6% ±12.6% vs. 67.3%±11.7%, p=0.030, respectively). The change in sperm motility after processing was significantly fewer in the non-pregnant cycles, both when the comparison was conducted by subtraction (post-pre) and division (post/pre). These relationships remained significant after adjusting for the female partner's age, anti-Müllerian hormone level, and number of pre-ovulatory follicles. According to a receiver operating characteristic curve analysis, an initial sperm motility of ≥72.5% was the optimal threshold value for predicting live birth after IUI. Conclusion: Initial sperm motility, rather than the motility of processed sperm or the degree of change after preparation, predicted live birth after IUI procedures.

• Reproductive and Developmental Biology
• /
• v.35 no.1
• /
• pp.85-91
• /
• 2011

The objective of this study was to evaluated the efficiency on sperm cryosurvival and ability of in vitro fertilization using Triladyl and Lactose Egg-Yolk(LEY) as extenders for cryopreservation of separated sperm by 65% percoll in miniature pig. Sperm viability was measured with SYBR-14/PI double stained sperm by flow cytometry. Ability on embryo cleavage rate and blastocyst development were observed by in vitro fertilization after frozen-thawing of sperm separated by 65% percoll. The experimental groups were designed that separated sperm by 65% percoll with Triladyl (ST) or LEY(SL) and unseparated sperm with Triladyl(UT) or LEY(UL) for cryopreservation. As a results, the viability was significantly(p<0.05) higher in ST(55.1%), SL(63.1%), UL(58.8%) than UT(38.2%) group. Sperm viability in SL(63.1%) group was significantly(p<0.05) higher than other experimental groups. On the other hand, embryo cleavage rate was significantly(p<0.05) higher in ST(79.1%), SL(83.2) than UT(74.1) and UL(75.7%) groups at 96h after in vitro fertilization. Blastocyst development was also significantly(p<0.05) higher in ST(21.5%), SL(20.9%) than UT(17.0%) and UL(18.8%) groups. In conclusion, cryopreservation of miniature boar sperm separated by 65% percoll were beneficial to viability and capacity on in vitro fertilization.

• Korean Journal of Animal Reproduction
• /
• v.3 no.1
• /
• pp.56-60
• /
• 1979

This study was carried out to find the difference between the naturaly born sex rtio among 1,242 head of pigs(120 litters) at Swine Farm, Cheil Sugar Co. and B-body a, pp.arance from their semen, and to find the conception rates which were inseminated to 40 sows with sperm from the anode and cathode after electrophoresis of boar semen. In order to the electrophoretic separation, the semen was placed into the platimum loop electrodes(105 cc) at room temperature for 30 minutes with D.C. 3V. and 350${\mu}$A. constant. The sperm fluorescent staining method was performed in accordance with Bhattacharya's(1970) method. The spermatozoa were observed through a Olympus Vanox microscope(made in Japan) using exciter filter with I heat barrier HPO 120. The results obtained were summarized as follows: 1. The natural sex ratio of 1,242 piglets(120 litters) which were born at Swine Farm, Cheil Sugar Co. was 50%, and B-body a, pp.arance of its boar semen were 49.24%. 2. With electrophoretic separation, the anode and cathode attracted 65.5${\pm}$5.03% and 29.89${\pm}$4.29% of B-body bearing sperm, respectively. 3. After electrophoresis of boar sperm, they were inseminated to 40 sows with sperm from anode and cathode. The conception rate was 92.5%.

• Clinical and Experimental Reproductive Medicine
• /
• v.49 no.3
• /
• pp.196-201
• /
• 2022

Objective: This prospective consecutive study investigated the variation in sperm DNA fragmentation (SDF) in multiple semen samples from patients with cancer. Methods: Eighty-one patients with various cancers underwent multiple semen collections on 3 consecutive days for sperm cryopreservation prior to cancer treatment. A commercial Halosperm kit was used to measure SDF. Within- and between-subject coefficients of variation were estimated via random-effects analysis of variance to assess the consistency of semen parameters and SDF. Intraclass correlation coefficients (ICCs) were calculated to assess the magnitude of the between-subject component of variance relative to the total variance. Results: The volume of semen in the day-2 and day-3 samples was significantly lower compared with the day-1 sample. Most parameters showed high ICC values, suggesting that within-subject fluctuations were small relative to the between-subject variability. The highest ICC values were identified for the SDF (ICC, 0.68; 95% confidence interval [CI], 0.45-0.84) and semen volume (ICC, 0.67; 95% CI, 0.45-0.84). Conclusion: Our findings showed that repeated ejaculates from patients with cancer had stable SDF levels.

• Korean Journal of Animal Reproduction
• /
• v.26 no.3
• /
• pp.275-289
• /
• 2002

The objective of this study was to develop an in vitro assessment of sperm fertilizing capacity of bulls and investigate the factors influencing sperm function and characteristics of frozen-thawed bovine spermatozoa. in vitro fertilization (IVF), the evaluation of motility and normal morphology, HOST (hypoosmotic swelling test), Ca-ionophore induced acrosome reaction, luminol and lucigenin-dependent chemiluminescence for the measurement of reactive oxygen species (ROS), the measurement of malondialdehyde formation for the analysis of lipid peroxidation (LPO), and the evaluation of DNA fragmentation using the method of 747-mediated nick end labelling (TUNEL) by flow cytometry were performed in frozen-thawed bovine spermatozoa. Correlations between the rates of fertilization, blastocyst formation after IVF and the values of respective assays were investigated. 1. IVF rate and blastocyst formation rate averaged 64.4% and 34.3% for spermatozoa from high -fertility bull group and averaged 18.5% and 6.2% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. Sperm motility and percentage acrosome reaction averaged 79.0% and 66.2% for spermatozoa from high-fertility bull group and averaged 40.7% and 22.9% for spermatozoa from low-fertility bull group, respectivitely. There were not different between two bull groups. 2. Luminol depenent chemiluminescence, LPO and DNA fragementation averaged 6.4, 2.0 nmol and 2.6% from spermatozoa from high-fertility bull group and averaged 6.5, 3.1 nmol and 7.4% for spermatozoa from low-fertility bull group, respectively. There were significantly different between two bull groups. There was no significant difference in lucigenin dependent chemiluminescence between two bull groups. 3. Fertilization rate was positively correlated with motility and the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between fertilization rate and the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. 4. Blastocyst formation rate was positively correlated with the rate of Ca-ionophore induced acrosome reaction, but negatively correlated with the frequency of luminol-dependent chemiluminescence, the rate of LPO, and the percentage of sperm with DNA fragmentation. There was no correlation between blastocyst formation rate and motility, the percentage of swollen spermatozoa, normal morphology, and the frequency of lucigenin-dependent chemiluminescence. In conclusion, these data suggest that ROS significantly impact semen quality. The assays of this study may provide a basis fur improving in vitro assessment of sperm fertilizing capacity.