Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

Effect of ${\alpha}$-Tocopherol Treatment and Freezing for In Vitro Bovine Embryo Production in Korean Native Cows

한우 체외수정란 생산을 위한 ${\alpha}$-Tocopherol 첨가와 동결 효과

  • 정진우 (농협중앙회 한우개량사업소) ;
  • 손준규 (농촌진흥청 국립축산과학원) ;
  • 최창용 (농촌진흥청 국립축산과학원) ;
  • 서상원 (농촌진흥청 국립축산과학원) ;
  • 최진석 (농촌진흥청 국립축산과학원) ;
  • 조인철 (농촌진흥청 국립축산과학원) ;
  • 한상현 (농촌진흥청 국립축산과학원) ;
  • 강태영 (제주대학교 수의학과) ;
  • 강민수 (제주대학교 수의학과) ;
  • 김덕임 (농협중앙회 한우개량사업소) ;
  • 김현종 (농촌진흥청 국립축산과학원) ;
  • 조상래 (농촌진흥청 국립축산과학원)
  • Received : 2011.09.07
  • Accepted : 2011.09.19
  • Published : 2011.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

The aim of this study was to evaluate the effect of ${\alpha}$-tocopherol on blastocysts development and subsequent cryosurvival of the vitrification. The ${\alpha}$-tocopherol(0, 100, 200, 400 ${\mu}M$) was added in to culture medium for the bovine embryos. The blasocysts from the ${\alpha}$-tocopherol and untreated control groups were then frozen-thawed, and their cryosurvival was assessed by in vitro culture for 48 h. There were no differences in the overall cleavage rate($56.14{\pm}4.66$, $58.18{\pm}4.70$, $62.97{\pm}6.86$ and $51.17{\pm}7.28$) among four treatment groups. However, in blastocyst development and total cell number were significantly higher in ${\alpha}$-tocopherol 200 ${\mu}M$ ($38.60{\pm}7.12$; $106.33{\pm}3.50$) to culture medium than other treatment groups($29.30{\pm}5.24$, $31.60{\pm}7.12$ and $26.37{\pm}4.18$; $101.36{\pm}5.12$, $97.27{\pm}2.87$, and $91.23{\pm}7.52$ respectively). Before and after vitrification, the total cell number and blastocyst development of embryo were significantly higher in July to August than September to October. In conclusion, addition of ${\alpha}$-tocopherol 200 ${\mu}M$ to in vitro bovine embryo culture medium was beneficial for improving embryo quality by decreasing the embryo damage blsstocysts cell number and improving the tolerance of the embryos to cryopreservation.

본 연구는 한우의 체외수정란 생산에서 체외성숙, 체외 배양시 배양액에 항산화제인 ${\alpha}$-Tocopherol의 첨가 농도에 따른 대조구와 처리구 난포란의 발달과 배반포 발생율 및 초자화 동결 후 생존성에 미치는 영향을 구명하고자 실험을 실시하였다. ${\alpha}$-Tocopherol 첨가 농도에 따른 분할율에서는 7~10월까지 비교한 결과, ${\alpha}$-tocopherol 200 ${\mu}M$ 첨가된 처리구가 대조구 및 타 처리구보다 높은 경향을 보였으나, 유의적 차이는 없었다. 배반포 발생율에서는 8~10월에서 ${\alpha}$-tocopherol 200 ${\mu}M$ 첨가된 처리구 $38.60{\pm}7.12%$가 대조구 및 타 처리구들보다 유의적으로 높게 (p<0.05) 나타났다. 초자화 동결 전 후의 총세포수는 ${\alpha}$-tocopherol 200 ${\mu}M$ 처리구에서 $115.80{\pm}6.61$개, $106.33{\pm}3.50$개로서, 다른 대조구 및 타 처리구보다 높은 세포수를 보였다. 월별에 따른 수정란의 배반포 발생율 및 동결 전후의 총세포수에서는 7~8월이 9~10월보다 유의적으로 높게 (p<0.05) 나타났다.

Keywords

References

  1. Basile JR, Chebel RJ, Basile LF (1998): Effect of season on embryo transfer in superovulated Holstein cows with PMSG. Br J Vet Res Anim Sci 35:257- 259.
  2. Bize I, Santander G, Cabello P, Driscoll D, Sharpe C (1991): Hydrogen peroxide is involved in hamster sperm capacitation in vitro. Biol Reprod 44:398- 403. https://doi.org/10.1095/biolreprod44.3.398
  3. Carlson JC, Wu XM, Sawada M (1993): Oxygen radicals and the control of ovarian corpus luteum function. Free Radic Biol Med 14:79-84. https://doi.org/10.1016/0891-5849(93)90511-R
  4. Carvalho RV, Del Campo MR, Palasza AT, Planteb Y, Mapletofta RJ (1996): Survival rates and sex ratio of bovine IVE embryos frozen at different developmental stages on day 7. Theriogenology 45:489- 498. https://doi.org/10.1016/0093-691X(95)00385-L
  5. Christensen LG (1991): Use of embryo transfer in future cattle breeding schemes. Theriogenology 35: 141-149. https://doi.org/10.1016/0093-691X(91)90153-5
  6. Ciocca DR, Oesterreich S (1993): Biological and clinical implications of heat shock protein 27000 (Hsp- 27). JNCI. J Natl Cancer Inst 85(19):1558-1570. https://doi.org/10.1093/jnci/85.19.1558
  7. Crosby IM, Gandolfi F, Moor RM (1988): Control of protein synthesis during early cleavage of sheep embryos. J Reprod Fertil 82:769-775. https://doi.org/10.1530/jrf.0.0820769
  8. Forsberg EJ (2005): Commercial applications of nuclear transfer cloning: three examples. Reprod Fertil and Develop 17(2):59-68. https://doi.org/10.1071/RD04114
  9. Gibson JP, Smith C (1986): Technology and animal breeding; Application in livestock production. 3rd world Cong. on Genetics Applied to Livestock Prod 12:96-105.
  10. Jeong YW, Park SW, Hossein MS, Kim S, Kim JH, Lee SH, Kang SK, Lee BC, Hwang WS (2006): Antiapoptotic and embryotrophic effects of alpha-tocopherol and L-ascorbic acid on porcine embryos derived from in vitro fertilization and somatic cell nuclear transfer. Theriogenology 66:2104-2112. https://doi.org/10.1016/j.theriogenology.2006.06.007
  11. Joenje H (1989): Genetic toxicology of oxygen. Mutation Research 1219:193-208.
  12. Jones DP (1985): Oxygen supply In : Oxidative Stress, London: Academic Press. 167-170.
  13. Kontush A, Finckh B, Karten B, Kohlschutter A, Beisiegel U (1996): Antioxidant and prooxidant activity of alpha-tocopherol in human plasma and low density lipoprotein. J Lipid Res 37:1436-1448.
  14. Kuwayama M (1995): Vitrification of IVMFC bovine embryos at various developmental stages and of different quality. Theriogenology 43:257-257. https://doi.org/10.1016/0093-691X(95)92411-2
  15. Lohuis MM (1997): Strategy for diary cattle improvement utilizing MOTE in anada. Anim Genetic and Breed 1:224-226.
  16. Long CR, Dovrinsky JR, Johnson LA (1999): In vitro production of pig embryos: comparisons of culture media and boars. Theriogenology 51:1375-1390. https://doi.org/10.1016/S0093-691X(99)00081-3
  17. Miyazaki T, Sueoka K, Dharmarajan AM, Atlas SJ, Bulkley GB, Wallach EE (1991): Effect of inhibition of oxygen free radical on ovulation and progesterone production by the in-vitro perfused rabbit ovary. J Reprod Fert 91:207-212. https://doi.org/10.1530/jrf.0.0910207
  18. Monty Jr DE, Racowsky C (1987): In vitro evaluation of early embryo viability and development in summer heat stressed, superovulated dairy cows. Therigenology 28:451-465. https://doi.org/10.1016/0093-691X(87)90250-0
  19. Nakao H, Nakasuji 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. https://doi.org/10.1016/0093-691X(90)90536-3
  20. Quinn P, Gaye M (2005): The effect of oxygen on the development of preimplantation mouse embryos in vitro. J Experi Zoo 206(1):73-80.
  21. Rall WF, Wood MJ, Kirby C, Whittingham DG (1987): Developmentof mouse embryos cryopreserved by vitrification. J Reprod Fertil 80:499-504. https://doi.org/10.1530/jrf.0.0800499
  22. Rodgers RJ, Lavranos TC, Rodgers HF, Young FM, Vella CA (1995): The physiology of the ovary: maturation of ovarian granulosa cells and a novel role for antioxidants in the corpus luteum. J Steroid Biochem Mol Biol 53:241-246. https://doi.org/10.1016/0960-0760(95)00054-4
  23. Ruane J (1988): Review of the use of embyro transfer in the genetic improvement of dairy cattle. Anim Breed 56:437-446.
  24. Smith C (1984): Genetic improvement of livestock, using nucleus breeding units. World Anim Rev 65: 2-10.
  25. Sreenan JM (1988): Embyro transfer: its uses and recent development. Veterinary Record 122:624-629. https://doi.org/10.1136/vr.122.26.624
  26. Sturman H, Oltenacu EA, Foote RH (2000): Importance of inseminating only cows in estrus. Theriogenology 53(8):1657-1667. https://doi.org/10.1016/S0093-691X(00)00305-8
  27. Tatemoto H, Sakurai N, Muto N (2000): Protection of porcine oocytes against apoptotic cell death caused by oxidative stress during In vitro maturation: role of cumulus cells. Biol Reprod 63:805. https://doi.org/10.1095/biolreprod63.3.805
  28. Thibodeaux JK, Del Vecchio RP, Hansel W (1993): Role of platelet-derived growth factor in development of in vitro matured and in vitro fertilized bovine embryos. J Reprod Fertil 98:61-66. https://doi.org/10.1530/jrf.0.0980061
  29. Thiyagarajan B, Valivittan K (2009): Ameliorating effect of vitamin E on in vitro development of preimplantation Buffalo embryos. J Assist Reprod Genet 26:217-25. https://doi.org/10.1007/s10815-009-9302-1
  30. Thompson JG, Partridge RJ, Houghton FD, Cox CI, Leese HJ (1996): Oxygen uptake and carbohydrate metabolism by in vitro derived bovine embryos. J Reprod Fertil 106:299-306. https://doi.org/10.1530/jrf.0.1060299
  31. Voelkel SA, Hu YX (1992): Direct transfer of frozen- thawed bovine embryos. Theriogenology 37:23- 37. https://doi.org/10.1016/0093-691X(92)90245-M
  32. Wales RG (1975): Maturation of the mammalian embryo: biochemical aspects. Biol Reprod 12:66-81. https://doi.org/10.1095/biolreprod12.1.66
  33. Wang B, Zhou J (2003): Specific genetic modifications of domestic animals by gene targeting and animal cloning. Reprod Biol Endo 1:103. https://doi.org/10.1186/1477-7827-1-103
  34. 손동수, 김일화, 류일선, 연성흠, 서국현, 이동원, 최선호, 박수봉, 이충섭 (2000): 젖소 MOET scheme의 추진을 위한 수정란 생산 및 이식. 한국수정란이식학회지 15(1):57-65.
  35. 조상래, 최창용, 김현종, 최선호, 손동수 (2010): 한우수정란의 동결보존 후 발달 효율 비교. 한국수정란이식학회지 23(3): 223-227.
  36. 최영진 (2001): 돼지 체외 수정란의 체외발육에 대한 항산화제의 첨가효과. 강원대학교 석사학위논문.