한국가금학회지 (Korean Journal of Poultry Science)

  • 제40권3호
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  • Pages.171-178
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  • 2013
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  • 1225-6625(pISSN)
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  • 2287-5387(eISSN)
한국가금학회 (The Korean Society of Poultry Science)
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닭 정액 동결 시 동결 보호제가 정액 성상에 미치는 영향

Effects of Kinds of Cryoprotectants on the Characteristics of Frozen Fowl Semen

  • 최진석 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 신단비 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 고응규 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 도윤정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 변미정 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 박수봉 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 성환후 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김현 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 공일근 (경상대학교 축산학과) ;
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장)
  • Choi, Jin Seok (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Shin, Dan-Bi (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Ko, Yeoung-Gyu (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Do, Yoon-Jung (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Byun, Mijeong (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Park, Soo-Bong (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Seong, Hwan-Hoo (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Kim, Hyun (Animal Genetic Resources Station, National Institute of Animal Science, RDA) ;
  • Kong, Il-Keun (Department of Animal Science, Division of Applied Life Science, Institute of Agriculture and Life Science, Graduate School of Gyeongsang National University) ;
  • Kim, Sung Woo (Animal Genetic Resources Station, National Institute of Animal Science, RDA)
  • 투고 : 2013.07.09
  • 심사 : 2013.07.30
  • 발행 : 2013.09.30
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초록

본 연구는 닭 정액 동결 보존을 위하여 동결 보호제로 7%의 DMA, DMF 그리고 7.5%의 MA를 이용하여 동결 융해 후 정자의 생존율, 정상 첨체율 및 미토콘드리아의 활성도를 FACS를 이용하여 평가하고, 수정률과의 상관관계를 조사하고자 수행하였다. 닭 정액을 동결한 결과, DMF를 사용하였을 때 생존율은 $52.1{\pm}5.52%$로 DMA와 MA에 비해 유의적으로 높았으며(P<0.05), DMA $46.94{\pm}5.06%$, MA $36.56{\pm}4.66%$순으로 나타났다(P<0.05). 첨체막이 파열되고 죽은 정자는 이와 반대로 MA를 이용하여 동결한 정액이 $61.16{\pm}1.86%$로서 가장 높았고, DMA $47.48{\pm}1.9%$ DMF $36.56{\pm}1.42%$순으로 유의적 차이를 보였다(p<0.05). 정자 미토콘드리아 막이 손상되지 않은 생존 정자는 DMF를 이용하여 동결한 정액에서 $52.68{\pm}1.07%$로 가장 높게 나타났고, DMA $44.46{\pm}1.04%$, MA $38.36{\pm}1.88%$순으로 유의적 차이를 나타냈다(p<0.05). 7% DMF를 사용하여 동결하였을 때 수정률은 유의적인 차이가 없었지만, 생존율 및 미토콘드리아 활성도가 유의적으로 가장 높았으며, 정상 첨체율이 유의적으로 높았다. 이는 DMF를 이용한 닭 정자 동결법이 정자의 세포막과 첨체의 손상에 미치는 영향이 가장 낮은 동결 보호제인 것으로 사료된다.

The purpose of this study was to evaluate the sperm viability, normal acrosome and mitochondrial activity in the frozen-thawed fowl semen by different cryoprotectants. The experiment was carried out on 10 sexually adult roosters of Ogye. The semen was collected twice a week and pooled semen was diluted 1:1 EK extender containing no cryoprotectant at $5^{\circ}C$. After equilibration for 30 minutes, diluted chicken semen was diluted 1:1 extender containing either 7% dimethylacetamide (DMA), 7% dimethylformamide (DMF) or 7.5% methylacetamide (MA) at final concentration and was put in 0.5 mL plastic straws and frozen for 30 minutes by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, followed by plunging into liquid nitrogen. Frozen semen was thawed in water bath at $5^{\circ}C$ for 2 minutes. For cytometric analysis, the frozen-thawed semen was diluted with EK extender to a final concentration of 90 million spermatozoa per mL. Sperm membrane integrity was evaluated as SYBR-14 and propidium iodide (PI). Acrosome integrity was assessed with fluorescein isothiocyanate-labeled PSA and PI. The percentage of mitochondrial function was estimated by using Rhodamine123 (R123) and PI. In conclusion, freezing rooster semen by using 7% DMF as cryoprotectant was significantly highest in rates of survival and mitochondrial function while its rate of damage of acrosome was significantly lowest. As a result, DMF is the cryoprotectant that has the lowest influences on sperm membranes and acrosome integrity. Therefore it could be used for freezing method of animal genetic conservation method for poultry diversity.

키워드

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피인용 문헌

  1. Effect of cryoprotectants and thawing temperatures on chicken sperm quality 2017, https://doi.org/10.1111/rda.13070
  2. Effect of N-Methylacetamide Concentration on the Fertility and Hatchability of Cryopreserved Ogye Rooster Semen vol.41, pp.1, 2014, https://doi.org/10.5536/KJPS.2014.41.1.21
  3. The Study of Estimation of Chromatin Abnormality of Ogye Rooster Sperm and Activity by Diff-Quik Staining Method vol.42, pp.2, 2015, https://doi.org/10.5536/KJPS.2015.42.2.109