Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Effects of Methyl-Beta-Cyclodextrin (MBCD) on Cold Shock and Membrane Cholesterol Quantity during the Freezing Process of Miniature Pig Spermatozoa

미니돼지 정액 동결 시 Methyl-Beta-Cyclodextrin (MBCD)이 냉각 충격과 막 콜레스테롤량에 미치는 영향

  • Lee, Sung-Young (College of Animal Life Sciences, Kangwon National University) ;
  • Lee, Yong-Seung (College of Animal Life Sciences, Kangwon National University) ;
  • Cheong, Hee-Tae (School of Veterinary Medicine, Kangwon National University) ;
  • Yang, Boo-Keun (College of Animal Life Sciences, Kangwon National University) ;
  • Park, Choon-Keun (College of Animal Life Sciences, Kangwon National University)
  • 이성영 (강원대학교 동물생명과학대학) ;
  • 이용승 (강원대학교 동물생명과학대학) ;
  • 정희태 (강원대학교 수의과대학) ;
  • 양부근 (강원대학교 동물생명과학대학) ;
  • 박춘근 (강원대학교 동물생명과학대학)
  • Received : 2011.08.15
  • Accepted : 2011.08.22
  • Published : 2011.09.30
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Abstract

This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until $5^{\circ}C$ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.

Keywords

References

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