• Title, Summary, Keyword: Boar Spermatozoa

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Association Study Analysis of Cluster-of-Differentiation Antigen 9 (CD9) Gene Polymorphism (g.358A>T) for Duroc Boar Post-thawed Semen Motility and Kinematic Characteristics

  • Cho, Eun-Seok;Sa, Soo-Jin;Kim, Ki-Hyun;Lee, Mi-Jin;Ko, Jun-Ho;Kim, Young-Ju;Seol, Kuk-Hwan;Hong, Joon-ki;Kim, Kwang-Sik;Kim, Yong-Min;Woo, Jae-Seok
    • Journal of Embryo Transfer
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    • v.30 no.2
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    • pp.109-114
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    • 2015
  • Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.358A>T) of cluster-of-differentiation antigen 9 (CD9) gene reported to be significant association with MOT. Also, CD9 gene was expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen. This study was conducted to evaluate the pig SNP (g.358A>T) of CD9 gene as a positional controlling for semen parameters of post-thawed boar semen. To results, the g.358A>T SNP of the CD9 gene was significantly associated with the traits such as MOT, curve linear velocity, straight line velocity, average path velocity and amplitude of lateral head displacement. Particularly, the g.358A>T SNP significantly has the highest association with MOT and animals with AA genotype (p<0.001). Therefore, we suggest that the g.358A>T in the intron 6 region of the porcine CD9 may be used as a molecular marker for Duroc boar Post-thawed semen quality, although its functional effect was not defined yet.

In Vitro Production of Pig Embryos using Intracytoplasmic Injection of Flow Cytometry Sorted Boar Spermatozoa

  • Kim, Dae-Young;Hyun, Sang-Hwan;Lee, Eun-Song
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.275-281
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    • 2008
  • The ability to preselect the sex of piglets is advantageous in the pig industry. The objective of this study was to examine the feasibility of using intracytoplasmic sperm injection (ICSI) with sorted spermatozoa to produce piglets with a preselected sex. Pig embryos were produced by ICSI of frozen X- and Y-sperm that had been separated by flow cytometry. The developmental competence of the embryos was investigated in vitro and in vivo. The populations of X- and Y-spermatozoa were 52.7% and 47.3%, respectively in our samples. The in vitro development of ICSI embryos was enhanced by longer of in vitro maturation of oocytes ($44{\sim}48\;h$ vs. $40{\sim}43\;h$). Their cleavage ($65{\sim}70%$) and blastocyst formation ($9{\sim}12%$) rates were not significantly different between male and female ICSI embryos, or between sorted and unsorted sperm-derived embryos. One pregnancy was established in a recipient that was transferred with 110 female ICSI embryos, but the pregnancy was terminated on Day 89 of gestation. Our results suggest that the separation X- and Y-spermatozoa by flow cytometric sorting can be a useful tool in combination with ICSI for the production of pig embryos and piglets of preselected sex.

A Study on the Natural Sex Ratio and Fertility of Galvanized Boar Semen (돼지의 자연성비와 정자의 전기분이에 의한 수태성적에 관한 연구)

  • 이용빈;오봉국;권종국;서국성;정영철;오성종
    • Korean Journal of Animal Reproduction
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    • v.3 no.1
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    • pp.56-60
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    • 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%.

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Association Study Analysis of Phospholipase C Zeta (PLCz) Gene Polymorphism (g.158T>C) for Duroc Boar Post-Thawed Semen Motility and Kinematic Characteristics (PLCz 유전자의 유전적 다형성(g.158T>C)과 두록 동결정액의 운동학적 특성과의 연관성 분석)

  • Sa, Soo-Jin;Lee, Mi-Jin;Kim, Ki-Hyun;Woo, Jae-Seok;Ko, Jun-Ho;Kim, Young-Ju;Cho, Eun-Seok
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.137-142
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    • 2015
  • Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.158T>C) of phospholipase C zeta (PLCz) gene reported to be significant association with MOT. This study was conducted to evaluate the PLCz gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.158 T>C SNP of PLCz was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.0001, p=0.0002, p<0.0001 and p<0.0001, respectively). Therefore, we suggest that the intron region of the porcine PLCz, may be used as a molecular marker for Duroc boar post-thawed semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

Effects of Green Tea (Camellia sinensis) Extract Supplementation at Different Dilution Steps on Boar Sperm Cryopreservation and in vitro Fertilization

  • Park, Sang-Hyoun;Jeon, Yubyeol;Yu, Il-Jeoung
    • Journal of Veterinary Clinics
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    • v.35 no.2
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    • pp.39-45
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    • 2018
  • We evaluated the effects of green tea extract (GTE) supplementation at different dilution steps on boar sperm freezing and in vitro fertilization. Sperm intracellular hydrogen peroxide ($H_2O_2$), motility, viability, acrosome integrity and morphology were determined. In addition, sperm IVF parameters (penetration and monospermy) and glutathione (GSH) levels of presumptive zygotes (PZs) were evaluated. Semen was diluted in lactose egg yolk (LEY) and cooled at $5^{\circ}C$ for 3 h (first dilution step) and then diluted in LEY with 9% glycerol and maintained at $5^{\circ}C$ for 30 min (second dilution step). Four experimental groups were compared: first and second dilution steps without GTE (control), first dilution step with GTE (Step 1), second dilution step with GTE (Step 2) and first and second dilution step with GTE (Step 1+2). The spermatozoa were frozen in nitrogen vapor. Higher sperm motility, viability and acrosome integrity after thawing were observed in Step 1, Step 2 and Step 1+2 groups compared with the control (P < 0.05). Lower $H_2O_2$ level was observed in Step 1+2 compared with control and Step 1 (P < 0.05). For IVF, matured oocytes were co-cultured with spermatozoa frozen according to the experimental groups. GSH levels of PZs were significantly higher in Step 2 and Step 1+2 than in control and Step 1 (P < 0.05) without a significant difference in IVF parameters. In conclusion, supplementation with GTE in both first and second dilution steps during the freezing process resulted in better boar sperm cryopreservation and might be beneficial for further embryo development.

Maintenance of Sperm Characteristics and In vitro Developmental Rate of Embryos against Oxidative Stress through Antioxidants in Pig

  • Jang, H.Y.;Kong, H.S.;Oh, J.D.;Park, B.K.;Yang, B.K.;Jeon, G.J.;Lee, H.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.3
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    • pp.340-345
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    • 2008
  • Oxidative stress is one of the major causes of failure of in vitro storage of boar semen. Reactive oxygen species (ROS) are one of the important mediators of oxidative stress during in vitro storage of boar semen. Our study examined the effects of taurine on sperm characteristic and on in vitro developmental embryos during in vitro storage of boar semen for 7 days. Semen was randomly aliquoted into 3 centrifuge tubes and treated with different concentrations of taurine (25-100 mM). The characteristics of boar sperm were analyzed for motility by light microscopy, viability by using a Makler counting chamber and membrane integrity by a hypoosmotic swelling test (HOST). The percentages of motile spermatozoa in taurine groups after 5 days were significantly higher compared to the control. Sperm viability in the control was lower than in taurine groups after 7 days irrespective of different taurine concentration. In the hyoosmotic swelling test (HOST), significantly higher results were obtained in taurine groups after 3 days. Also, the developmental rates of IVM/IVF porcine embryos from semen treated with pyruvate and taurine were significantly increased when compared with the control (p<0.05). These results indicate that supplementation of taurine as an antioxidant in boar semen extender can improve the semen quality.

Effects of Panax ginseng, zearalenol, and estradiol on sperm function

  • Gray, Sandra L.;Lackey, Brett R.;Boone, William R.
    • 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.

Effects of Reactive Oxygen Species (ROS) on Sperm Function and Plasminogen Activator Activity in Porcine Spermatozoa

  • Sa, Soo-Jin;Park, Chun-Keun;Kim, In-Cheul;Lee, Seung-Hoon;Kwon, Oh-Sub;Kim, Myung-Jick;Cho, Kyu-Ho;Kim, Du-Wan;So, Kyoung-Min;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.185-191
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    • 2010
  • Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin and may participate in mammalian fertilization. Although correlations have been reported between reactive oxygen species (ROS) and sperm function, the relationship between PA activity and ROS is unknown. We determined the effects of ROS on sperm function and PA activities in boar spermatozoa preincubated under the X-XO system. When spermatozoa were treated with the X+XO group, a significant increase (p<0.05) was observed in the percentage of acrosome reacted spermatozoa compared with that of the control group. However, when antioxidants were added to the medium with X+XO, the rate of acrosome reaction tended to decrease. Also, a significantly lower percentage of acrosome reacted spermatozoa was observed in the X+XO+catalase group at 6 hr of incubation compared with that of X+XO group. The density of malondialdehyde (MDA) was higher in the X+XO group than in different treatment groups. In another experiment, incubation of spermatozoa in medium with X+XO was associated with a significant (p<0.05) increase in activity of tPA-PAI and tPA compared with the control group. Antioxidants decreased the increased activity of tPA-PAI and tPA by preincubation in the X-XO system. Also, a significantly lower (p<0.05) activities of tPA-PAI and tPA were observed in the X+XO+catalase group compared with the X+XO group. No significant differences, however, were observed in the activity of uPA. These results suggest that the increase of acrosome reaction by the X-XO system resulted in increase of PAs activity in the sperm incubation medium.

Characteristics of Phthalate Esters-exposed Boar Sperm during Boar Semen Storage (돼지 정액을 보관하는 동안 phthalate esters에 노출된 정자의 특성)

  • Lee, A-Sung;Lee, Sang-Hee;Lee, Seunghyung;Yang, Boo-Keun
    • Journal of Life Science
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    • v.29 no.4
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    • pp.395-401
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    • 2019
  • Phthalate is a chemical endocrine disrupter and interfere with the action of hormones, estrogens, androgens and thyroid hormones. It also affect cardiovascular, metabolic, immune and reproductive system in the human and animals. Curcumin is antioxidant, anti-inflammatory activity and -cancer properties in the human. We studied whether phthalates damage viability, mitochondrial activity and membrane integrity of sperm in boar semen. We also treated curcumin with/without phthalates in the boar semen. Fresh boar semen was treated with phthalates and/or curcumin for examining sperm characteristics. Sperm characteristics, sperm motility, viability, mitochondrial activity, and membrane integrity were determined during storage of boar semen. Sperm motility and viability in dose-dependent manner decreased by di-n-butyl phthalate (DBP), mono-n-butyl phthalate (MBP) and di-2-ethylhexyl phthalate (DEHP, p<0.05). Phthalates also decreased mitochondrial activity and membrane integrity of sperm (p<0.05). However, sperm motility and viability were higher than untreated-curcumin when DBP, MBP and DEHP treated with a curcumin in boar semen (p<0.05). Mitochondrial activity and membrane integrity of sperm were higher in DBP- and MBP-treated semen with curcumin (p<0.05). In conclusion, phthalates can damage sperm viability and quality during the boar semen storage, and curcumin may protect the boar sperms from phthalates during storage term.