• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.247-252
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• 2005

The purpose of this study was to assess sperm quality during in vitro storage of miniature-pig semen in order to determine which extender should be used and how extender can be diluted for in vitro storage of miniature-pig semen. Freshly ejaculated miniature-pig's semen was diluted with same volumes of Beltsville Thawing Solution (BTS), Androhep, Modena, Mulberry III and modified-Modena extenders. Sperm quality was evaluated by examining viability, motility, abnormality, acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining. Sperm motility decreased with storage period prolonged and differences among BTS, Androhep, Modena and Mulberry III were apparent On Day 1, approximately 80% of the sperm were motile, but motility decreased to $40\%$ at Day 7. During the 7 days of storage, sperm survival in modified-Modena B extender was higher than another extenders. However, it was not differ significantly among other extenders. The percentage of F and B patterns were not differ significantly among the extenders. However, F pattern in modified-Modena B extender was slightly higher until 3 days of storage than that of Modena extender, modified-Modena A extender and modified-Modena C extender. The percentage of AR patterns in modified-Modena B extender was slightly lower, but did not differ significantly among other extenders. The results of present study suggest that modified-Modena B was effective as new extender for in vitro storage of miniature-pig semen.

• Korean Journal of Animal Reproduction
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• v.9 no.2
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• pp.153-157
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• 1985

Ejaculated and epidiymal goat spermatozoa were preserved for 0, 6, 12 adn 18 h, and 0 and 18 h in a semi-aerobic condition at 20-$25^{\circ}C$, and preincubated for 5-6 h in a CO2 incubator in m-KRB solution. Then they were preincubated at different concentrations (3-5, 25-48 and 105-190$\times$107/ml), and ability of penetration into zona-free hamster eggs in vitro was examined. When ejaculated spermatozoa were preincubated in m-KRB solution after presservation for 12 and 18 h, 12 and 29% of zona-free eggs were penetrated, and only 4% of eggs were penetrated by epididymal spermatozoa which were preincubated after preservation for 18 h. When spermatozoa were preincubated at a low concentration, the penetration rates were very low. But when the sperm concentration during preincubation was 25-48 and 105-190$\times$107/ml, the penetration rates increased to about 30%.

• Journal of Veterinary Clinics
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• v.17 no.2
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• pp.420-430
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• 2000

These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

• Korean Journal of Animal Reproduction
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• v.14 no.4
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• pp.297-302
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• 1990

This study was carried out to investigate the effect of heparin, CSA and PC12 on sperm motility and acrosome reaction in bovine fresh and frozen semen which were washed and incubated in mTALP, and also the effect of heparin-pretreatment on motility and acrosome reaction in mTALP, and also the effect of heparin-pretreatment on motility and acrosome reaction of sperm treated with PC12, and the results obtained were as follows : 1. When fresh sperm was once washed and then incubated for 15 minutes in mTALP containing heparin 1, heparin 2, CSA and PC12, the percent of motile sperm of PC12 was significantly lower than that of control, heparin 1, heparin 2 and CSA. But the percent of acrosomereacted sperm of PC12 was signifciantly higher than that of control, heparin 1, heparin 2, and CSA. 2. When frozen sperm was once washed and then incubated for 15 minutes in mTALP containing heparin 1, heparin 2, CSA and PS12, there was no significant difference in the percent of motile sperm among treatments, but the percent of acrosome-reacted sperm of PC12 was signifciantly higher than that of heparin 2, and there was no significant difference in the percent of acrosome-reacted sperm among control, heparin and CSA. 3. When fresh sperm was twice washed and then incubated for 15 minutes in mTALP containing heparin and PC12, there was no significant differrence in the percent of motile sperm among treatments, but the percent of acrosome-reacted sperm of PC12 was significantly higher than that of control and heparin. When the sperm was incubated for 120 minutes, the percent of motile sperm of PC12 was significantly lower than that of control and heparin, but the percent of acrosome-reacted sperm of PC12 was significantly higher than that of control and heparin. 4. When fresh sperm was twice washed and preincubated in mTALP containing heparin for 0, 15, 120, and 240 minutes, and then incubated with PC12 for 15 minutes, there was no significant difference in the perce수 of motile sperm among treatments, but the percent of acrosome-reacted sperm of 120 and 240 minutes was significantly higher than that of 0 and 15 minutes.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.241-249
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• 2017

Plasma membrane hyperpolarization associated with activation of $Ca^{2+}$-activated $K^+$ channels plays an important role in sperm capacitation during fertilization. Although Slo3 (slowpoke homologue 3), together with the auxiliary ${\gamma}^2$-subunit, LRRC52 (leucine-rich-repeat-containing 52), is known to mediate the pH-sensitive, sperm-specific $K^+$ current KSper in mice, the molecular identity of this channel in human sperm remains controversial. In this study, we tested the classical $BK_{Ca}$ activators, NS1619 and LDD175, on human Slo3, heterologously expressed in HEK293 cells together with its functional interacting ${\gamma}^2$ subunit, hLRRC52. As previously reported, Slo3 $K^+$ current was unaffected by iberiotoxin or 4-aminopyridine, but was inhibited by ~50% by 20 mM TEA. Extracellular alkalinization potentiated hSlo3 $K^+$ current, and internal alkalinization and $Ca^{2+}$ elevation induced a leftward shift its activation voltage. NS1619, which acts intracellularly to modulate hSlo1 gating, attenuated hSlo3 $K^+$ currents, whereas LDD175 increased this current and induced membrane potential hyperpolarization. LDD175-induced potentiation was not associated with a change in the half-activation voltage at different intracellular pHs (pH 7.3 and pH 8.0) in the absence of intracellular $Ca^{2+}$. In contrast, elevation of intracellular $Ca^{2+}$ dramatically enhanced the LDD175-induced leftward shift in the half-activation potential of hSlo3. Therefore, the mechanism of action does not involve pH-dependent modulation of hSlo3 gating; instead, LDD175 may modulate $Ca^{2+}$-dependent activation of hSlo3. Thus, LDD175 potentially activates native KSper and may induce membrane hyperpolarization-associated hyperactivation in human sperm.

• Journal of Embryo Transfer
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• v.13 no.3
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• pp.291-300
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• 1998

This study was carried out to elucidate the effect of progesterone, estradiol 17 beta and cholesterol in follicular fluid on sperm chemotaxis for fertilization. By inducing swim-up migration through sucrose layer into bMSS containing progesterone, estradiol 17 beta and/or cholesterol, their effects on sperm migration and sperm movement were examined. And the results obtained were as follows; 1. Progesterone inhibited sperm migration and movement, but significantly attracted capacitated-sperm at the level of 50 $\mu$g/ml. 2. Estradiol 17 beta inhibited sperm migration and movement, but didn＇t significantly inhibit migration of capacitated-sperm at the level of 10$\mu$g/ml. 3. Cholesterol significantly stimulated sperm migration and movement at the level of 50$\mu$g/ml, but didn＇t attact capacitated-sperm. 4. Progesterone and estradiol 17 beta reduced the effect of cholesterol stimulating sperm migration and movement. But estradiol 17 beta and cholesterol didn＇t reduce the effect of progesterone attracting capacitated-sperm. In conclusion, progesterone of 50$\mu$g/ml in bMSS attracted the capacitated-sperm, cholesterol of 50$\mu$g/ml stimulated sperm migration and movement, but estradiol 17 beta of 10$\mu$g/ml didn＇t affect sperm swim-up separation.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.261-268
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• 1994

Immatured bovine follicular oocytes added with serum, hormones, granulosa cells and bovine oviduct epithelium cells were fertilized in vitro after in vitro maturation. In vitro maturation and early development capacity were examined and IVF-derived embryos were transferred and to recipients and effects of sperm treatment on in vitro capacitation were investigated. The rate of in vitro maturation was improved when they were co-culutred with granulosa cells in the TCM199 medium added with 10% FCS and hormones. The percentage of acrosome reaction was not differed between sperm treatments and sperm of above 25% under-went AR during 30 min preincubation with caffeine and heparin. The cleavage rate of oocytes in vitro fertilized in TCM199 medium added with 10% FCS and hormones, GC or BOEG higher than that in medium with 10% FCS and GC. But the rate was not significantly different between GC and BOEG The cleavage of rate oocytes cultured in medium containing serum, hormones and BOEG was 80.2% and more embryos were developed to Blastocyst (17.3%). The selected embryos were transferred to 9 recipients by surgical or nonsurgical method but did not result in pregnancy.

• Journal of Ginseng Research
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• v.40 no.3
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• pp.251-259
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• 2016

Background: Estrogen signaling pathways are modulated by exogenous factors. Panax ginseng exerts multiple activities in biological systems and is classified as an adaptogen. Zearalenol is a potent mycoestrogen that may be present in herbs and crops arising from contamination or endophytic association. The goal of this study was to investigate the impact of P. ginseng, zearalenol and estradiol in tests on spermatozoal function. Methods: The affinity of these compounds for estrogen receptor (ER)-alpha and beta ($ER{\alpha}$ and $ER{\beta}$)-was assessed in receptor binding assays. Functional tests on boar spermatozoa motility, movement and kinematic parameters were conducted using a computer-assisted sperm analyzer. Tests for capacitation, acrosome reaction (AR), and chromatin decondensation in spermatozoa were performed using microscopic analysis. Results: Zearalenol-but not estradiol ($E_2$)- or ginseng-treated spermatozoa-decreased the percentage of overall, progressive, and rapid motile cells. Zearalenol also decreased spontaneous AR and increased chromatin decondensation. Ginseng decreased chromatin decondensation in response to calcium ionophore and decreased AR in response to progesterone ($P_4$) and ionophore. Conclusion: Zearalenol has adverse effects on sperm motility and function by targeting multiple signaling cascades, including $P_4$, $E_2$, and calcium pathways. Ginseng protects against chromatin damage and thus may be beneficial to reproductive fitness.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.4
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• pp.220-229
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• 2021

Lonicera caerulea (Honey berry, HB) has been used in medical treatment in Russia, Japan, China and Korea. It has high level of vitamin C and polyphenolics. Polyphenolics can improve anti-inflammatory effect and prevent cancer, diabetes mellitus type 2. Also, Vitamin C is a representative anti-oxidant. however, it is still unknown what effect it will have on the oxidation stress of the reproductive system. In previous studies, ROS can be produced when it is exposed to heat stress and has negative effect on sperm's maturation, capacitation, hyperactivation, acrosome reaction and fusion of egg and sperm. Therefore, the purpose of this study is to investigate the antioxidant effects of L. Caerulea on the sperm and mice. At first, it conducted using ICR mouse (n = 20) for 4 weeks. There are four groups of mice (n = 5 per group). Also, L. Caerulea was taken by oral gavage. Group I (control) kept at 23℃-27℃ and administer D.W (0.5 mL/day), Likewise, Group II (HB) kept at room temperature but gave HB (250 mg/kg, 0.5 mL/day), Group III (HB + HS) received heat stress (40℃) using hyperthermia induction chamber and gave HB at same dose. and Group IV (HS) exposed heat stress only. Mainly, we showed degree of gene expression using Western blot in SOD, HSP 70, 17β-HSD and Real-time PCR. It can find correlation between intracellular activity like steroid hormone, apoptosis under ROS and antioxidant activity of L. Caerulea.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.229-237
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• 1999

The influence of ascorbic acid (Asc) and ferrous sulfate (Fe$^{2＋}$) on capacitation, acrosome reaction and fertility in vitro was investigated in boar frozen-thawed spermatozoa with or without preincubation. The addition of 0-1.0 mM Fe$^{2＋}$ to sperm suspensions during preincubation increase acrosome reaction (P＜0.05) and oocyte penetration. These increase are also associated with addition of 0~0.5 mM Asc, but the penetration rates were higher in those without than with sperm preincubation. The addition of 0.1 mM Asc than 0.5 mM in medium with Fe$^{2＋}$ were significantly (P＜0.05) higher on acrosome reaction at 2 h after sperm preincubation. No significant differences, however, were observed in penetration rates among the concentrations of Asc. On the other hand, when preincubation medium containing the Asc was supplemented with 0.1mM Fe$^{2＋}$, the percentage of spermatozoa acrosome-reacted were significantly (P＜0.05) higher than in medium wothout Fe$^{2＋}$, on the contrary, the penetration rate was significantly (P＜0.05) low during in-vitro fertilization. These findings indicate some apparent effects of Fe$^{2＋}$ or Asc addition on acrosome reaction and the fertilizing potential by sperm preincubation.