Comparison of Oocyte Recovery Rates between Prepubertal and Adult Korean Native Goats

미성숙 및 성숙 재래 산양의 과배란 처리에 의한 난자의 회수율 비교

  • Yun, Yun-Jin (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology) ;
  • Park, Hee-Sung (Department of Animal Science and Biotechnology, Gyeongnam National University of Science and Technology)
  • 윤윤진 (경남과학기술대학교 동물생명과학과) ;
  • 박희성 (경남과학기술대학교 동물생명과학과)
  • Received : 2011.08.02
  • Accepted : 2011.08.20
  • Published : 2011.09.30

Abstract

This study was conducted to examine whether efficiency of oocyte production from superovulated prepubertal goats. Fifteen prepubertal and twenty adult goats, maintained in a pen under natural day length and fed hay ad libitum, were pretreated with progestagen implanted CIDR for 10 days. Superovulation treatment of the goats received twice daily intramuscular injection of a total of 70 mg FSH for 3 days from Day 8 of CIDR. All the gonadotrophin treated goats were injected with 10 mg $PGF_2{\alpha}$ on Day 8 and 400~600 IU hCG in the afternoon on Day 10. Oocytes were recovered by follicle aspiration or oviduct flushing at 35 to 40 h after hCG injection through mid-ventral incision. The in vivo matured oocytes was activated by ionomycin (5 min) and 6-DMAP (3.5~4 h). The activated oocytes were cultured in mSOF medium containing 0.8% BSA at 38.5$^{\circ}C$ in an atmosphere of 5% CO$_2$, 5% O$_2$, 90% N$_2$ for 7~8 days. There was no significant difference in the mean number of CL and in vivo matured and follicular oocytes recovered. But, quality of I + II grade follicular oocytes was lower (p<0.05) in the prepubertal goat (25.0%) than the adults (52.4%). The same results were also observed in the cleavage and blastocyst rate of activated oocytcs. The cleavage and blastocyst rate from prepubertal derived oocytes were lower (p<0.05) in the prepubertal goat (54.5%, 23.3%) than the adult goat (86.8%, 46.6%). Considering overall these results, we suggest that maturation of donor goats is a major factor affecting recovered oocytes quality and in vitro development of activated goat oocytes.

본 연구는 효율적인 재래 산양의 난포란을 확보하기 위하여 미성숙 산양에 과배란 처리를 실시하여 체내 성숙 난자(배란된 난자) 및 난포란을 회수하여 공란산양의 성숙 여부가 난자의 회수율과 단위 발생란의 체외발달율에 미치는 영향을 조사하였다. 미성숙 산양의 과배란 처리에 의한 체내 성숙 난자의 회수율에 있어서 과배란 처리 후 두당 황체수는 13.1${\pm}$6.5개로서 성숙 산양의 8.8${\pm}$2.4개와 차이가 없었다(p<0.05). 산양의 두당 회수율은 마성숙 산양이 9.l${\pm}$6.8개로서 성숙 산양의 9.8${\pm}$2.5개와 차이가 없었다. 공란산양으로부터 체내 성숙 난자 회수 후 난소의 난포로부터 채란한 난포란의 회수율에 있어서 산양 두당 난포 수는 미성숙 산양이 7.9${\pm}$6.5개로서 성숙 산양의 8.8${\pm}$2.4개와 차이가 없었다. 이들 황체로부터 회수한 난포란 수도 미성숙 산양이 5.6${\pm}$6.1개로서 성숙 산양의 6.3${\pm}$2.3개와 차이가 없었다. 회수한 난포란의 등급에 있어서 1+II 등급 난포란은 미성숙 산양이 25.0%로서 성숙 산양의 52.4%보다는 낮았다(p<0.05). 4등급의 비율은 미성숙 산양이 39.3%로서 성숙 산양의 19.8%보다는 높았다(p<0.05). 미성숙 산양으로부터 회수한 체내 성숙 난자를 단위 발생을 유도하였을 때 분할율은 54.5%로서 성숙 산양으로부터 회수한 단위 발생 난자의 분할율 86.8%보다는 낮았다(p<0.05). 배반포기로의 발달율에 있어서도 미성숙 산양이 23.3%로서 성숙 산양의 46.6%보다는 유의적 (p<0.05)으로 낮았다. 이상의 결과를 미루어 볼 때 미성숙 재래 산양으로부터 난자의 생산은 난자의 확보 측면에서는 한 방법이 될 수 있으나, 회수란의 질 저하에 대한 문제는 공란 산양의 보다 적합한 연령, 실험 시기, 성선자극 호르몬의 양 및 공시 산양의 관리 등 앞으로 보다 세밀한 검토가 필요한 것으로 생각된다.

Keywords

References

  1. Anguita B, Vandaele L, Mateusen B, Maes S and Van Soom A. 2007. Developmental competence of bovine oocytes is not related to apoptosis incidence in oocytes, cumulus cells blastocysts. Theriogenology 67:537-549. https://doi.org/10.1016/j.theriogenology.2006.09.004
  2. Armstrong DT, Holm P, Irvine B, Petersen BA, Stubbings RB, McLean D, Stevens G and Seamark RF. 1992. Pregnancies and live birth from in vitro fertilization of calf oocytes collected by laparoscopic follicular aspiration. Theriogenology 38:667-678. https://doi.org/10.1016/0093-691X(92)90029-Q
  3. Armstrong DT, Irvine BJ and Earl CR. 1994. In vitro fertilization of follicular oocytes from juvenile lambs and their developmental competence in vitro and in vivo. Biol. Reprod. Abst. 50:189.
  4. Baldassarre H and Karatzas CN. 2004. Advanced assisted reproduction technologies (ART) in goat. Anim. Reprod. Sci. 82-83:255-26. https://doi.org/10.1016/j.anireprosci.2004.04.027
  5. Catt SL. 2002. Effects of cytoplasmic transfer from adult to prepubertal derived oocytes on the in vitro production of ovine blastocysts. Reproduction Abst. 28:59.
  6. Earl CR, Irvine B and Armstrong DT. 1994. Development of techniques for the production of viable embryos from six to seven week old lambs. In proceedings of the Australian Society of Animal production Abst. 20:428.
  7. Ikeda K and Takahashi Y. 2003. Comparison of maturational and developmental parameters of oocytes recovered from prepubertal and adult pigs. Reprod. Fertil. Dev. 15:215-221. https://doi.org/10.1071/RD02077
  8. Izquierdo D, Villamediana P, Lopez-Bejar M and Paramio MT. 2002. Effect of in vitro and in vivo culture on embryo development from prepubertal goat IVM-IVF oocytes. Teriogenology 57:1431-1441. https://doi.org/10.1016/S0093-691X(02)00647-7
  9. Jimenez-Macedo AR, Paramio MT, Anguita B, Morato R, Romaguera R, Mogas T and Izquierdo D. 2007. Effect of ICSI and embryo biopsy on embryo development and apoptosis according to oocyte diameter in prepubertal goats. Theriogenology 67:1399-1408. https://doi.org/10.1016/j.theriogenology.2007.03.003
  10. Kelly JM, Kleeman DO and Walker SK. 2003. A comparison of the in vitro development of cow and calf oocytes. Theriogenology Abst. 59:449.
  11. Kelly JM, Kleeman DO and Walker SK. 2005. Enhanced efficiency in the production of offspring from 4- to 8-week-old lambs. Theriogenology 63:1876-1890. https://doi.org/10.1016/j.theriogenology.2004.09.010
  12. Kennedy JP, Worthington CA and Cole ER, 1974. The post-natal development of the ovary and uterus of the Merino lamb. Reprod. Fertil. 36:275-282. https://doi.org/10.1530/jrf.0.0360275
  13. Khatir H, Lonergan P, Carolan C and Mermillod P. 1996. Prepubertal bovine oocyte: A negative model for studying oocyte developmental competence. Mol. Reprod. Dev. 45:231-239. https://doi.org/10.1002/(SICI)1098-2795(199610)45:2<231::AID-MRD17>3.0.CO;2-3
  14. Kochhar HPS, Wu B, Morris LHA, Buckrell BC, Pollard JW, Basrur PK and King WA. 2002. Maturation status, protein synthesis and developmental competence of oocytes derived from lambs and ewes. Reprod. Dom. Anim. 37:19-25. https://doi.org/10.1046/j.1439-0531.2002.00326.x
  15. Koeman JK, Keefer CL, Baldassarre H, Lane M, Garnder DK and Downey BR. 2003. Developmental competence of prepubertal and adult goat oocytes cultured in semi-defined media following laparoscopic recovery. Theriogenology 60:879- 889. https://doi.org/10.1016/S0093-691X(03)00090-6
  16. Leoni GG, Ledda S, Bogliolo L, Succu S, Rosati I, bebbere D, Pintus PP and Naitana S. 2004. Ovine prepubertal oocyte shows alterate gene expression and low developmental competence. Reprod. Fertil. Dev. Abst. 16:240.
  17. Leoni GG, Succu S, Satta V, Paolo M, Bogliolo L, Bebbere D, Spezzigu A, Madeddu M, Berlinguer F, Ledda S and Naitana S. 2009. In vitro production and cryotolerance of prepubertal and adult goat blastocyst obtained from oocytes collected by laparoscopic oocyte-pick-up (LOPU) after FSH treatment. Reprod. Fertil. Dev. 21:901-908. https://doi.org/10.1071/RD09015
  18. Marchal R, Feugang JM, Perreau C, Venturi E and Mermillod P. 2000. Developmental competence of prepubertal and adult swine oocytes: Birth of piglets from in vitro produced blastocyst. Theriogenology Abst. 53:361.
  19. Martino A, Mogas T, Palomo MJ and Paramio MT. 1994. Meiotic competence of prepubertal goat oocytes Theriogenology 41:969-980. https://doi.org/10.1016/0093-691X(94)90512-H
  20. Mogas T, Palomo MJ, Izquierdo MD and Paramio MT. 1997. Developmental capacity of in vitro matured and fertilized oocytes from prepubertal and adult goats. Teriogenology 47:1189-1203. https://doi.org/10.1016/S0093-691X(97)00099-X
  21. Morton KM, Catt SL, Maxwell WM and Evans G. 2005. An efficient method of ovarian stimulation and in vitro embryo production from prepubertal lambs. Reprod. Fertil. Dev. 17:701-706. https://doi.org/10.1071/RD04105
  22. Morton KM. 2008. Developmental capabilities of embryos produced in vitro from prepubertal lamb oocytes. Reprod. Domest. Anim. 43:137-143.
  23. O'Brien JK, Catt SL, Ireland KA, Maxwell WMC and Evans G. 1997. In vitro and in vivo developmental capacity of oocytes from prepubertal and adult sheep. Theriogenology 47:1433-1443. https://doi.org/10.1016/S0093-691X(97)00134-9
  24. Palma GA, Tortonese DJ and Sinowatz F. 2001. Developmental capacity in vitro of prepubertal oocytes. Anat. Histol. Embryol. 30:295-300. https://doi.org/10.1046/j.1439-0264.2001.00324.x
  25. Ptak G, Loi P, Dattena M, Tischner M and Cappai P. 1999. Offspring from one-month-old lambs: Studies on the developmental capability of prepubertal oocytes. Biol. Reprod. 61:1568-1574. https://doi.org/10.1095/biolreprod61.6.1568
  26. Ptak G, Tischner M, Bernabo N and Loi P. 2003. Donor dependant developmental competence of oocytes from lambs subjected to repeated hormonal stimulation. Biol. Reprod. 69:278-285. https://doi.org/10.1095/biolreprod.102.011312
  27. Revel F, Mermillod P, Peynot N, Renard JP and Heyman Y. 1995. Low development capacity of in vitro matured and fertilized oocytes from calves compared with that of cows. J. Repord. Fertil. 103:115-120. https://doi.org/10.1530/jrf.0.1030115
  28. Romaguera R, Morato R, Jimenez-macedo AR, Izquierdo D, Catala M, Roura M, Paramio MT, Palomo MJ, Mogas T and Izquierdo D. 2010. Oocyte secreted factors improve embryo developmental competence of COCs from small follicles in prepubertal goats. Theriogenology 74:1050-1059. https://doi.org/10.1016/j.theriogenology.2010.04.035
  29. Velilla E, Izquierdo D, Rodriguez-Gonzalez E, Lopez-Bejar M, Vidal F and Paramio MT. 2004. Distribution of prepubertal and adult goat oocyte cortical granules during meiotic maturation and fertilisation: Ultrastructural and cytochemical study. Mol. Reprod. Dev. 68:507-514. https://doi.org/10.1002/mrd.20111
  30. Urdaneta A, Jimenez AR, Paramio MT and Izquierdo D. 2004. Cysteamine, glutathione and ionomycin treatments improve in vitro fertilization of prepubertal goat oocytes. Zygote 12: 277-284. https://doi.org/10.1017/S0967199404002874
  31. 박희성, 이지삼, 정장용. 2000. 한국 재래 산양의 난포란의 회수 와 체외수정에 관한 연구. 한국수정란이식학회지 15:287-293.
  32. 박희성, 정수영, 김태숙, 이명열, 진종인, 홍승표, 이지삼, 김충희.2004. 재래 산양의 과배란처리에 있어서 회수시간이 난자의회수율에 미치는 영향. 한국수정란이식학회지 19:113-119.
  33. 박희성, 김태숙, 정수영, 박준규, 이지삼, 정장용. 2006. 공핵세포 및 발정동기화가 복제 재래산양 생산에 미치는 영향.한국수정란이식학회지 21:137-146.
  34. 정수영, 박희성. 2007. 복제 산양(진순이)의 체세포 핵이식에의한Re-Cloning에 관한 연구. 한국수정란이식학회지 22:89-95.