Korean Journal of Animal Reproduction (한국가축번식학회지)

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

Studies on the Developmental Rate of Oocyter Obtained fly Intracytoplasmic Sperm Injection with Epididymal Spermatozoa in Domestic Dogs

개 난자에 부고환 정자로 ICSI후 배양하였을 때 체외발생율에 관한 연구

  • 김상근 (충남대학교 수의과대학) ;
  • 이동수 (충남대학교 수의과대학) ;
  • 이만희 (워싱턴대학교 세포생리학과)
  • Published : 2002.06.01
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Abstract

The objective of this study was to determine the developmental competence of in vitro matured oocytes after intracytoplasmic sperm injection(ICSI) with epididymal spermatozoa. The ovaries were obtained from slaughtered small species dogs. Oocytes matured in vitro for 24 hrs were fertilized by ICSI with epididymal spermatozoa. After ICSI, one group of oocytes was activated with 2.0 mM dimethylaminopurine or 7% ethanol for 5 min. and second group was not activated. The follicular oocytes were cultured in synthetic oviductal fluid(SOF) and TCM-199 medium containing hormones and 10% FCS for 24~48 hrs in a incubator with 5% $CO_2$ in air at 38.5$^{\circ}C$. 1. Results of IVM showed that the percentage of oocytes reaching MII after 24 h and 48 hrs of incubation were significantly higher(p<0.05) after culture with 48 hrs(9/30, 30.0%) than that after culture with 24hrs(a/30, 26.7%). 2. Results of IVM showed that the percentage of oocytes reaching MII after 48 hrs of incubation were significantly higher(p<0.05) after culture with SOF media(10/30, 30.3%) than TCM-199 media (7/30, 23.3%). 3. The rate of cleavaged embryos to blastocyst obtained by ICSI treated activation oocytes was significantly higher(p<0.05) than that of nonactivation oocytes(5/16, 25.0% vs 1/13, 5.0%). 4. The rates of development of cleavaged embryos to blastocyst obtained by ICSI treated sperm of fresh, epididymal and frozen-thawed epididymal were 8/18(44.43%), 5/16(31.3%), 2/14(14.3%), respectively. and these values of frozen-thawed epididymal sperm injection were lower than fresh sperm injection.

본 연구는 소형견의 불임 해결과 번식효율 증진을 위해 소형견 난소 난포로부터 채취한 난자를 활성화 처리후 부고환 정자로 ICSI시켰을 때 체외발생율을 조사하기 위하여 수행하였다. 1. 난포란을 회수 후 24, 48시간 배양하였을 때 배양시간에 따른 GV, MI, MII로의 체외발생율은 각각 14/30(46.7%), 2/30(6.7%), 8/30(26.7%)였고 48시간 배양 시간에 따른 GV, MI, MII로의 체외발생율은 각각 l1/30(36.7%), 3/30(10.0%), 9/30(30.0%)였다. 2. 난포란을 회수 후 48시간 배양하였을 때 배양액에 따른 MII로의 체외발생율은 SOF액(10/30, 30.3%)에서의 배양이 TCM-199액(7/30, 23.3 %)보다 높은 체외발생율을 나타냈다. 3. 활성화 처리 난자에 부고환 정자로 ICSI를 하였을 때 상실배와 배반포로의 체외발생율은 각각 3/16(18.8%), 4/16(25.0%)로서 비활성화 처리 난자군의 3/13(23.1%), 1/13(7.7%)에 비해 높은 체외발생율을 나타냈다. 4. 활성화 처리 난자에 신선정자, 부고환 정자 및 동결 융해한 부고환 정자로 ICSI를 하였을 때 체외발생율은 각각 8/18(44.4%), 5/16(31.3%), 2/14(14.3%)로서 동결 부고환 정자 처리군은 신선정자 처리군에 비해 낮은 체외발생율을 나타냈다.

Keywords

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