• Title/Summary/Keyword: Sperm chromatin structure assay

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Antioxidant Supplementation Enhances the Porcine Semen Preservation Capacity

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.39 no.1
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    • pp.7-11
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    • 2015
  • Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.

Effect of Storage Times on Sperm Function, Sperm Chromatin Structure Assay (SCSA) and Correlations Between Fertility and SCSA in Boars (액상 정액의 보존 기간이 정자 기능 및 정자염색질 구조 분석에 미치는 영향과 인공수정 분만율과의 상관관계)

  • 유재원;이주형;김인철;이일주;강 권;민동수;윤희진;윤종택;방명걸;류범용;정영채;김창근
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.785-796
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    • 2006
  • This study was designed to evaluate the changes in sperm motility, viability, HOST(hypo-osmotic swelling test), IVP(in vitro penetration), SCSA(sperm chromatin structure assay) during storage of liquid semen collected from boars with different farrowing rates using AI, and to find the relationship between boar fertility through AI and sperm diagnostic parameters during semen storage. The results of HOST were significantly decreased according to the increasing of in semen storage days and the results of IVP were significantly decreased at 3 days of semen storage (P<0.05). The %Red was significantly different among the >80%, 70󰠏80% and <70% farrowing rate group at semen storage day 6(P<0.05). The correlation coefficients between the %Red and farrowing rate were increased according to the semen storage. In conclusion, these results suggest that the sperm parameters evaluated in these studies may be useful indicators to predict the fertility of AI and evaluate the semen quality in boars.

Effect of Monothioglycerol on ROS Inhibition, Mitochondrial Activity, and DNA Integrity in Frozen-thawed Miniature Pig Sperm (Monothiolglycerol이 동결 융해 후 미니돼지 정자의 활성산소 억제, 미토콘드리아 활성 그리고 DNA Integrity에 미치는 영향)

  • Park, Soo-Jung;Kim, Dae-Young
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.265-271
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    • 2013
  • Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.