Korean Journal of Poultry Science (한국가금학회지)

The Korean Society of Poultry Science (한국가금학회)
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The Study of Estimation of Chromatin Abnormality of Ogye Rooster Sperm and Activity by Diff-Quik Staining Method

Diff-Quik 염색방법에 의한 오계 닭 정자의 염색질 이상과 운동성 추정에 관한 연구

  • Kim, Sung Woo (Animal Genetic Resources Station, National Institue of Animal Science, RDA) ;
  • Choi, Ahreum (Animal Genetic Resources Station, National Institue of Animal Science, RDA) ;
  • Choe, Changyong (Animal Genetic Resources Station, National Institue of Animal Science, RDA) ;
  • Kim, Dongkyo (Animal Genetic Resources Station, National Institue of Animal Science, RDA) ;
  • Seong, Hwan-Hoo (Animal Genetic Resources Station, National Institue of Animal Science, RDA) ;
  • Kim, Jae-Hwan (Animal Genetic Resources Station, National Institue of Animal Science, RDA) ;
  • Kim, Chongdae (Poutry Science Division, National Institute of Animal Science, RDA)
  • 김성우 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 최아름 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 최창용 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김동교 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 성환후 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김재환 (농촌진흥청 국립축산과학원 가축유전자원시험장) ;
  • 김종대 (농촌진흥청 국립축산과학원 자원개발부 가금과)
  • Received : 2015.03.24
  • Accepted : 2015.05.18
  • Published : 2015.06.19
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Abstract

Ogye rooster sperm chromatin status can be detected using well established sperm assays. In this paper, a simple and fast method to monitor rooster sperm chromatin status could be employed in field for assessment of chicken sperm quality. Using standard bright field microscope, Diff-Quik stains can be reproducibly, easily and routinely monitored with simple staining. The presence of abnormal chromatin staining of rooster sperm was determined by darker stain in head. In the fresh semen, the viabilities of three tested Ogye spermatozoa were 93.53%, 82.42% and 90.63% and normal chromatin rates were 87.96%, 74.25% and 85.10% respectively. However, after freezing, the rates of viability of thawed semen were reduced to 69.58%, 61.98% and 72.20% and normal chromatin rate also reduced to 58.91%, 48.49% and 63.34%. A significant correlation between live sperm and normal sperm nuclei was 0.875 in fresh semen and 0.513 in frozen semen. After incubation of sperm at $37^{\circ}C$ for 5min, the rates of viability, chromatin normality and sperm head activity were shown as $90.63{\pm}1.28%$, $82.44{\pm}8.09%$ and $66.68{\pm}10.29%$ in fresh semen. However, the rates of thawed semen were reduced to $67.92{\pm}7.55%$, $56.92{\pm}12.15%$ and 47.32{\pm}5.02%, respectively. The relationship between chromatin normality and sperm head movements in fresh and thawed semen were 0.564 and 0.540, respectively. With these results, the chicken sperm normality could be assessed by the Diff-Quik staining that could be used for chromatin status of sperm head and activated morphology of live spermatozoa, as a simple and rapid staining method.

본 연구에서는 육종이 필요한 종계장에서 슬라이드 위에 도말된 닭 정액시료를 간편하게 염색하여 정자의 염색질 이상성과 운동성을 유추할 수 있는 기법을 확립하고자 실시하였다. 오계 정자 도말 슬라이드를 Diff-Quik 시약으로 염색하여 관찰하면 신선정액에서는 정자 생존성이 93.53%, 82.42% 및 90.63%일 때, Diff-Quik 염색질의 정상도는 87.96%, 74.25% 및 85.10%로 관찰되었다. 동일한 시료를 동결하고 융해후 정액의 생존성은 조사하면 69.58%, 61.98% 및 72.20%로 관찰되었으며, 염색질의 정상도는 58.91%, 48.49% 및 63.34%로 관찰되었다. 융해된 동결정액에서 활력이 우수한 정자를 쉽게 관찰하기 위하여 정자를 HS-1 희석액으로 재 희석하고, $37^{\circ}C$에서 가온하여 도말하면 염색된 정자의 두부에서 활력이 우수한 정자의 비율을 간단히 유추할 수 있음을 보여주었다. 특히, 신선 정자에서 정상 염색질을 가진 정자의 비율과 생존성이 상관관계는 매우 높은 것으로 판단되며, 동결정자에서도 상관관계가 높다고 추정되었다. 이러한 결과는 Diff-Quik 염색방법으로 닭 정액의 품질을 정자의 염색질의 이상성 유무로 판단할 수 있음을 보여주고 있다. 특히 본 연구에서 제시된 방법은 닭 정자의 우수성을 판단하는데 유용할 것으로 판단되며, 닭 정액 시료를 준비할 때 준비한 도말슬라이드를 현미경적 관찰에 의하여 활성화된 정자의 비율과 정상 염색질을 가진 정자의 비율을 추정하는 방법을 제시하였으며, 이는 인공 수정에 필요한 현장에서 수컷 개체의 종축 이용성을 쉽게 판단할 수 있는 근거를 마련할 수 있음을 의미한다.

Keywords

References

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