한국가축번식학회지 (Korean Journal of Animal Reproduction)

한국동물번식학회 (The Korean Society of Animal Reproduction)
권호 표지

한우 수정란의 체외발육에 있어서 Aesculetin과 $O_2$농도의 영향

Effect of Aesculetin and $O_2$Concentrations on In Vitro Development of Preimplantation Embryo in Hanwoo (Korean Native Cattle)

  • Jang, H. Y. (College of Animal Resource Science, Kangwon National University) ;
  • Park, K. E. (Division of Animal Science, Faculty of Agriculture, Okayama University) ;
  • Kim, C. I. (College of Animal Resource Science, Kangwon National University) ;
  • Park, C. K. (College of Animal Resource Science, Kangwon National University) ;
  • H. T. Cheong (College of Animal Resource Science, Kangwon National University) ;
  • B. K. Yang (College of Animal Resource Science, Kangwon National University)
  • 발행 : 2002.03.01
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초록

본 연구는 aesculetin과 $O_2$농도가 체외에서 성숙 수정된 한우 초기배의 체외발육에 미치는 영향을 검토하였다. 실험을 위하여 체외수정 후 40~44시간에서 2~8세포로 발육한 초기배는 6일간 배양하였다. 그 결과, 실험 1에서 상실배와 배반포기배의 발육은 산소농도 20% 보다는 5%에서 유의 적으로 높게 나타났으며 (P＜0.05), 1 $\mu\textrm{g}$/$m\ell$의 aesculetin 첨가시 0, 5, 및 10 $\mu\textrm{g}$/$m\ell$ 첨가에 비하여 유의적 (P＜0.05)으로 높은 발육율을 나타냈다 실험 2에서, 배반포기 및 상실배 이상 발육한 초기배의 비율은 산소농도 20%보다는 5%에서 역시 유의적으로 더 높았으며 (P＜0.05), 1$\mu\textrm{g}$/$m\ell$의 aesculetin 또는 2.5mM taurine 첨가시 배반포기 및 상실배 이상 발육한 초기배의 비율은 다른 실험구에 비하여 유의적으로 높게 나타났다 (P＜0.05). 실험 3에서, aesculetin과 PDGF 동시 첨가 및 taurine과 EGF를 동시에 첨가한 경우 다른 실험구에 비하여 유의적으로 높은 발육율을 얻었으며 (P＜0.05), aesculetin 무첨가 보다는 첨가시 배반포기배의 세포수가 높게 관찰되었으나 배양조건에 관계없이 유의적인 차이는 인정되지 않았다. 한편, 실험 4에서 자연에서 추출한 것과 상품화된 aesculetin이 초기배의 발육에 미치는 영향을 검토한 결과 무첨가에 비해 유의적으로 높은 발육율을 나타냈으나 (P＜0.05), 자연에서 추출한 것 (71%)과 상품화 (70%)된 aescuietin 사이에 유의적인 차이는 인정되지 않았다. 또한 자연에서 추출한 것과 상품화된 aesculetin 첨가시 무첨가에 비해 배반포기배의 세포수가 높게 관찰되었으나 이들 사이에 의한 유의적인 차이는 없었다. 본 연구의 결과로부터, 5%의 산소농도하에서 aesculetin은 항산화제와의 공동첨가에 의해 한우 초기배의 체외발육을 증진시키는 것으로 나타났다.

The present study was examined effects of aesculetin and $O_2$concentrations on in vitro development of Hanwoo (Korean Native Cattle) embryos derived from in-vitro matured and fertilized (IVM-lVF) oocytes. The oocytes were cultured for the first 40~44 h after in vitro fertilization, then embryos of 2 to 8 cell stages were cultured under the different culture conditions for another 6 days. In experiment 1, the higher rates of morulae and blastocysts were produced in 5% $O_2$, than in 20% $O_2$(P＜0.05). There was significantly (P＜0.05) higher in embryos cultured with 1 $\mu\textrm{g}$/$m\ell$ than with 0, 5 and 10 $\mu\textrm{g}$/$m\ell$ of aesculetin. In experiment 2, the proportions of embryo developed with blastocysts and morulae plus blastocysts in 5% $O_2$, again was significantly (P＜0.05) higher in 20% $O_2$, during the culture with aesculetin and/or taurine. In the 5 and 20% $O_2$atmosphere, the inclusion of 1 $\mu\textrm{g}$/$m\ell$ aesculetin or 2.5mM taurine increased significantly (P＜0.05) the percentages of blastocysts and morulae plus blastocysts. In experiment 3, in medium with aesculetin plus PDGF and taurine plus EGF than other treatment groups, significantly (P＜0.05) higher developmental rates were obtained. Number of blastomere in balstocyst stage were also higher in medium with that than without aesculetin. However, there were no significant differences in all culture conditions. In experiment 4, the proportions of embryo developed to the morulae and blastocyst stages were significantly (P＜0.05) higher rates in medium with natural and commercial aesculetin than in control medium. No significant differences, however, were observed in between natural (71%) and commercial (70.0%) aesculetin. Number of blastomere in blastocyst stage were also higher in medium with natural and commercial aesculetin than in control medium. However, there was no effect on the number of blastomeres by these treatment. These data indicate that preimplatation embryos are very sensitive to condition that can cause oxygen concentration and show that efficiency role of aesculetin for improving bovine embryo development in vitro.

키워드

참고문헌

  1. Betterbed, B. and Wright, R. W. Jr. 1985. De-velopment of one-cell ovine embryos in two culture media under two gas atmospheres. Theriogenology 23:547-553
  2. Bulkely, G. B. 1987. Free radical-mediated reperfusion injury: a selective review. Br. J. Cancer (Supp!.) 55:66-73
  3. Craven, P. A., Pfanstiel, J. and DeRubertis, F. R. 1986. Role of reactive oxygen in bile salt stimulation of colonic epithelial proliferation. J. Clin. Invest. 77:850-859 https://doi.org/10.1172/JCI112382
  4. Fischer, B. and Bavister, B. D. 1993. Oxygen tension in the oviduct and uterus of the rhesus monkey, hamster and rabbit. J. Reprod. Fertil. 99:673-679 https://doi.org/10.1530/jrf.0.0990673
  5. Flood, M. R., Gage, T. L. and Bunch, T. D. 1993. Effect of various growth promoting factors on preimplantation bovine embryo de-velopment in vitro. Theriogenology 39:823-833 https://doi.org/10.1016/0093-691X(93)90421-Z
  6. Gandolfi, F., Brevini, T. A. L., Modina, S. and Lauria, A. 1991. Detection and characterization of a growth factor in bovine oviduct secretions. J. Reprod. Fertil. (Abstr.) 7:6 (Abs. 2)
  7. Keefer, C. L., Stice, S. L., Paprocki, A. M. and Golueke, P. 1994. In vitro culture of bovine IVM-IVF embryos:cooperative interaction among embryos and the role of growth factors. The-riogenology 41:1323-1331
  8. Larson, R. C., Ignotz, G. G. and Currie, W. B. 1991. Platelet derived growth factor (PDGF) initiates completion of the fourth cell cycle of bovine embryo development. J. Reprod. Fertil. (Abstr.) 7:6 (Abs. 1)
  9. Lee, E. S. and Fukui, Y. 1995. Effect of various growth factors in a defined culture medium on in vitro development of bovine embryos matured and fertilized in vitro. The-riogenology 44:71-83
  10. Li, J. and Foote, R. H. 1993. Culture of rabbit zygotes into blastocysts in protein-free medium with one to twenty percent oxygen. J. Reprod. Fertil. 98:163-167 https://doi.org/10.1530/jrf.0.0980163
  11. Liu, Z. and Foote, R. H. 1995. Development of bovine embryos in KSOM with added superox-ide dismutase and taurine and with five and twenry percent $O_{2}$. Biol. Reprod. 53:786-790 https://doi.org/10.1095/biolreprod53.4.786
  12. Liu, Z., Foote, R. H. and Yang, X. 1995. Development of early bovine embryos in co-culture with KOSM and taurine, superoxide dismutase or insulin. Theriogenology 44:741-750
  13. Mass, D. H. A., Storey, B. T. and Mastroianni Jr. L. 1976. Oxygen tension in the oviduct of the rhesus monkey (Macaca mulatta). Fertil. Steril. 27:1312-1317
  14. Nakao, H. and Nakatsuji, N. 1990. Effects of co-culture, medium components and gas phase on in vitro culture of in vitro matured and in vitro fertilized bovine embryos. Theriogenology 33:591-600
  15. Papaioannou, V. E. and Ebert, K. M. 1988. The preimplantation pig embryo: cell number and allocation to trophectoderm and inner cell mass of the blastocyst in vivo and in vitro. Develop-ment 102:793-803
  16. Sekiya, K., Okuda, H. and Arichi, S. 1982. Selective inhibition of platelet lipoxygenase by esculetin. Bilchim. Biophys. Acta. 713:68-72
  17. Tervit, H. R., Whittingham, D. G. and Rowson, L. E. A. 1972. Successful culture in vitro of ship and cattle ova. J. Reprod. Fertil. 30:493-497 https://doi.org/10.1530/jrf.0.0300493
  18. Voelkel, S. A. and Hu, Y. X. 1992. Effect of gas atmosphere on the development of one-cell bovine embryos in two culture systems. Therio-genology 37:1117-1131 https://doi.org/10.1016/0093-691X(92)90109-5
  19. Weiss, S. J. 1986. Oxygen, ischemia and in-flammation. Acta. Physiol. Scand. (Suppl) 548:9-37