Journal of Animal Science and Technology

Korean Society of Animal Sciences and Technology (한국축산학회)
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Effect of Amides as a Cryoprotectant on Quality of Frozen-thawed Sperm in Korean Jeju Black Bull

제주흑우 동결정액 제조 시 Amide 계열의 동결보호제가 동결 융해 후 정자의 성상에 미치는 영향

  • Oh, Shin-Ae (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA) ;
  • Choi, Sun-Ho (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA) ;
  • Ko, Min-Hee (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA) ;
  • Kang, Tae-Young (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA) ;
  • Cho, Sang-Rae (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA) ;
  • Ko, Moon-Suck (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA) ;
  • Oh, Young-Mi (Department of Veterinary Medicine, Jeju National University) ;
  • Cho, Won-Mo (Subtropical Animal Experiment Station, National Institute of Animal Science, RDA)
  • 오신애 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 최선호 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 고민희 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 강태영 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 조상래 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 고문석 (농촌진흥청 국립축산과학원 난지축산시험장) ;
  • 오영미 (국립제주대학교 수의과대학) ;
  • 조원모 (농촌진흥청 국립축산과학원 난지축산시험장)
  • Received : 2012.04.10
  • Accepted : 2012.04.23
  • Published : 2012.04.30
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Abstract

The objective of this study was to examine the effect of amides as a cryoprotectant for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing 5% dimethyl acetamide (DMA), 5% dimethyl formamide (DMF), 5% methyl formamide (MF) or 7% glycerol. Post-thawed sperm were evaluated for sperm motility, viability, acrosome integrity and membrane integrity. Post-thawed sperm motility was significantly higher (p<0.05) in glycerol and DMF ($64.00%{\pm}9.62$ and $59.00%{\pm}5.48$, respectively) than DMA and MF ($50.00%{\pm}3.24$ and $44.00%{\pm}4.18$, respectively). Sperm viability wassignificantly higher (p<0.05) in glycerol and DMF ($58.25%{\pm}7.35$ and $53.05%{\pm}3.77$, respectively) than others. However, for sperm motility and viability, there were no differences among glycerol and DMF. Also, swelling sperm ratio by hypo-osmetic selling test (HOST) was significantly increased (p<0.05) in glycerol and DMF treatments ($45.12%{\pm}25.08$ and $44.95%{\pm}8.58$, respectively). The percentage of capacitated sperm assessed by CTC staining, F pattern was lower (p<0.05) in DMF than others. B pattern was increased (p<0.05) in DMA, DMF and MF when compared with glycerol. AR pattern ratio was decreased (p<0.05) in glycerol and DMF when compared with DMA and MF. These results suggested that amides performed better and could be used as a cryoprotectant for semen freezing of Korean Jeju Black Bull.

본 연구는 희소 한우인 제주흑우의 정액 동결을 위하여 동결보호제로서 glycerol과 amide 계열의 동결보호제인 5%의 DMA, DMF 그리고 7%의 MF를 이용하여 동결 융해 후 정자의 운동성, 생존율, 정자막 온전성 및 정자의 첨체 양상의 변화 조사를 위하여 수행하였다. 제주흑우의 정액을 동결 한 결과 glycerol과 DMF를 사용하였을 때 운동성은 $64.00{\pm}9.62$와 $59.00{\pm}5.48$로 DMA와 MF에 비해 유의적으로 높았으며(p<0.05), 생존율 역시 glycero과 DMF 처리구에서 $58.25{\pm}6.63$과 $53.05{\pm}3.77$로 유의적으로 높게 나타났다(p<0.05). 그러나 glycerol과 DMF 처리구 간의 유의적 차이는 나타나지 않았다. 정자막 온전성 검사(HOST)에 있어서도 역시 glycerol과 DMF 처리에서 정자 미부의 팽창 비율 역시 $45.12{\pm}25.08$과 $44.95{\pm}8.58$로 유의적으로 높았다(p<0.05). 정자의 첨체막 양상 변화에 있어 F pattern의 비율이 MF 처리시 다른 처리구에 비하여 유의적으로 낮았으며(p<0.05), B pattern의 비율은 DMA, DMF 및 MF의 사용시 glycerol에 비하여 유의적으로 증가하였다(p<0.05). AR pattern의 비율에 있어서는 DMF를 제외한 amide의 사용은 F pattern의 감소와 조기 첨체반응에 의한 유의적인 AR pattern의 증가를 볼 수 있었다(p<0.0.5). 그러나 DMF의 사용은 glycerol 보다 AR pattern의 비율이 유의적으로 낮았다(p<0.05). 동결 융해 후 시간에 따른 정자의 생존율과 정자미부 팽창 비율에 있어서 융해 초기에는 glycerol에서 DMF 보다 높은 생존율과 정자 미부 팽창율을 나타냈으나, 1시간 이후부터는 DMF에서 glycerol 보다 높은 생존율과 정자 미부 팽창율을 나타내었다. 이러한 결과는 희소 가축의 생식세포 보존 및 유전자원 확립을 위한 중요한 자료가 될 것이며, 제주흑우 및 희소 가축의 안정적이고 효율적인 동결 정액 제조 연구에 있어 중요한 정보를 제공할 것으로 평가된다.

Keywords

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