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Effects of Zardaverine in Freezing Extender on Kinetic Characteristics of Post-Thawed Boar Sperm

동결보존액에 Zardaverine의 첨가가 동결-융해 후 돼지 정자의 운동학적 특성에 미치는 영향

  • Kim, Jeong A (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Cho, Eun Seok (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Jeong, Yong Dae (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Choi, Yo Han (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Hong, Jun Ki (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Kim, Young Sin (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Chung, Hak Jae (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Baek, Sun Young (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Sa, Soo Jin (Swine Science Division, National Institute of Animal Science, Rural Development Administration)
  • 김정아 (농촌진흥청 국립축산과학원 양돈과) ;
  • 조은석 (농촌진흥청 국립축산과학원 양돈과) ;
  • 정용대 (농촌진흥청 국립축산과학원 양돈과) ;
  • 최요한 (농촌진흥청 국립축산과학원 양돈과) ;
  • 홍준기 (농촌진흥청 국립축산과학원 양돈과) ;
  • 김영신 (농촌진흥청 국립축산과학원 양돈과) ;
  • 정학재 (농촌진흥청 국립축산과학원 양돈과) ;
  • 백선영 (농촌진흥청 국립축산과학원 양돈과) ;
  • 사수진 (농촌진흥청 국립축산과학원 양돈과)
  • Received : 2020.07.03
  • Accepted : 2020.09.04
  • Published : 2020.09.30
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Abstract

This study investigated the effect of Zardaverine supplementation in freezing extender, on kinetic characteristics of post-thawed boar sperm. Cryopreservation of boar sperm is an important technique of assisted reproductive technology and genetic resource banking. Although this technique is particularly useful, freeze-thaw cycles associated with sperm cryopreservation significantly reduce sperm quality. Semen from mature Duroc boars were collected and cryopreserved in freezing extenders (LEY) treated with varying concentrations of Zardaverine (0, 20, 50, 75, 100 𝜇M). The time-dependent kinetic characteristics of post-thawed spermatozoa were determined after thawing by applying computer-assisted sperm analysis (CASA). We observed that the motility immediately after thawing was significantly higher in 20 𝜇M stocks than in control (0 𝜇M) and the other treatments (p<0.05). Curvilinear velocity (VCL) in 0 𝜇M and 20 𝜇M stocks were significantly higher than the other treatment groups, except 75 𝜇M (p<0.05). Higher average path velocity (VAP) was obtained at 20 𝜇M as compared to 100 𝜇M, whereas amplitude of head lateral displacement (ALH) was significantly higher at 20 𝜇M than 50 𝜇M and 100 𝜇M (p<0.05). No differences were obtained for Straight-line velocity (VSL) and Linearity (LIN). In conclusion, our results indicate that Zardaverine improves the motility, VCL, VAP, and ALH of post-thawed boar sperm.

본 연구에서는 동결보존액에 대한 Zardaverine (phosphodiesterase inhibitor) 첨가가 돼지 동결-융해 정자의 운동학적 특성에 미치는 효과를 조사하였다. 돼지정액의 동결보존은 보조 번식기술 및 유전자원 장기보존을 위해 유용하게 이용되는 중요한 기술이다. 하지만 정자세포를 동결-융해하는 과정에서 발생하는 온도충격은 정자의 수정능력을 급격히 저하시킨다. 정액샘플은 성숙한 Duroc종 수퇘지로부터 채취했으며, lactose-egg yolk 동결보존액에 다양한 농도로 Zardaverine (0, 20, 50, 75 및 100 𝜇M)을 첨가하여 정액을 동결하였다. 융해 후 정자세포의 운동학적 특성 분석은 정자운동성자동분석기(CASA; computer-assisted sperm analysis)를 이용하였다. 그 결과, 융해 직후 정자의 운동성(MOT)은 타처리구에 비해 20 𝜇M 처리구에서 가장 높았다(p<0.05). Curvilinear velocity (VCL)은 0 𝜇M 과 20 𝜇M 처리구가 75 𝜇M 처리구를 제외한 다른 처리구들에 비해 유의적으로 높은 값을 보였다(p<0.05). Average path velocity (VAP)는 20 𝜇M 처리구가 100 𝜇M 처리구에 비해 유의적으로 높았으며(p<0.05) Amplitude of head lateral displacement (ALH)는 20 𝜇M 처리구가 50 𝜇M과 100 𝜇M 처리구에 비해 유의적으로 높았다(p<0.05). 이상의 결과를 종합하면, 동결용 보존액에 대한 Zardaverine 첨가가 동결-융해된 돼지 정자의 운동학적 특성에 긍정적인 영향을 미치는 것으로 사료된다.

Keywords

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