Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Improved Enucleation Efficiency of Pig Somatic Cell Nuclear Transfer by Early Denudation of Oocytes at 30 Hours of In Vitro Maturation

  • Song, Kil-Young (College of Veterinary Medicine, Seoul National University) ;
  • Hyun, Sang-Hwan (College of Veterinary Medicine, Chungbuk National University) ;
  • Lee, Eun-Song (School of Veterinary Medicine, Kangwon National University)
  • Published : 2007.12.31
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Abstract

Our goal was to examine the effects of early denudation on the enucleation efficiency and developmental competence of embryos following somatic cell nuclear transfer (SCNT) and parthenogenetic activation (PA). Oocytes were denuded following 30 h of in vitro maturation (IVM) and then cultured with (D+) or without (D-) their detached cumulus cells for additional $10{\sim}14$ h. Control oocytes were denuded after $40{\sim}44$ h of IVM. The size of the perivitelline space was larger at 40 h of IVM ($11.7{\sim}11.8{\mu}m$) than at 30 h ($8.9{\mu}m;$ p<0.01). The distances between the metaphase II (M II) plates and the polar bodies (PBs) were shorter in D+ ($19.4{\mu}m$) and D- oocytes ($18.9{\mu}m$) than in control oocytes ($25.5{\mu}m;$ p<0.01). Enucleation rates following blind aspiration at 40 h of IVM were higher (p<0.01) in D+ (92%) and D- oocytes (93%) compared to controls (82%). Early denudation did not affect oocyte maturation or the in vitro development of SCNT and PA embryos. When SCNT embryos from D+ oocytes were transferred to four gilts, pregnancy was established in two pigs, and one of them farrowed three live piglets. In conclusion, early denudation of oocytes at 30 h of IVM could improve the enucleation efficiency by maintaining the M II plate and the PB within close proximity and support the in vivo development of SCNT embryos to term.

Keywords

References

  1. Atabay EC, Martinez Diaz RA, Dochi S and Takahashi Y. 2001. Factors affecting enucleation rates of bovine and porcine oocytes after removal of cumulus cells by vortexing. J. Reprod. Dev., 47:365-371 https://doi.org/10.1262/jrd.47.365
  2. Bavister BD, Leibfried ML and Lieberman G. 1983. Development of pre implantation embryos of the golden hamster in a defined culture medium. Biol. Reprod., 28:235-247 https://doi.org/10.1095/biolreprod28.1.235
  3. Binor Z and Wolf DP. 1979. In-vitro maturation and penetra- tion of mouse primary oocytes after removal of the zona pellucida. J. Reprod. Fertil., 56:309-314 https://doi.org/10.1530/jrf.0.0560309
  4. Cibelli JB, Stice SL, Golueke PJ, Kane JJ, Jerry J, Blackwell C, Ponce de Leon FA and Robl JM. 1998. Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science, 280: 1256-1258 https://doi.org/10.1126/science.280.5367.1256
  5. Funahashi H and Day BN. 1993. Effects of the duration of exposure to hormone supplements on cytoplasmic maturation of pig oocytes in vitro. J. Reprod. Fertil., 98:179-185 https://doi.org/10.1530/jrf.0.0980179
  6. Hewitson L, Dominko T, Takahashi D, Martinovich C, Ramalho-Santos J, Sutovsky P, Fanton J, Jacob D, Monteith D, Neuringer M, Battaglia D, Simerly C and Schatten G. 1999. Unique checkpoints during the first cell cycle of fertilization after intracytoplasmic sperm injection in rhesus monkeys. Nat. Med., 5:431-433 https://doi.org/10.1038/7430
  7. Hyun SH, Lee GS, Kim DY, Kim HS, Lee SH, Kim S, Lee ES, Lim JM, Kang SK, Lee BC and Hwang WS. 2003. Effect of maturation media and oocytes derived from sows or gilts on the development of cloned pig embryos. Theriogenology, 59:1641-1649 https://doi.org/10.1016/S0093-691X(02)01211-6
  8. Ikeda K and Takahashi Y. 2001. Effects of maturational age of porcine oocytes on the induction of activation and development in vitro following somatic cell nuclear transfer. J. Vet. Med. Sci., 63:1003-1008 https://doi.org/10.1292/jvms.63.1003
  9. Isobe N, Maeda T and Terada T. 1998. Involvement of meiotic resumption in the disruption of gap junctions between cumulus cells attached to pig oocytes. J. Reprod. Fertil., 113: 167-172 https://doi.org/10.1530/jrf.0.1130167
  10. Kafi M, Mesbah F, Nili Hand Khalili A. Chronological and ultrastructural changes in camel (Came/us dromedaries) oocytes during in vitro maturation. Theriogenology, 2005; 63:2458-2470 https://doi.org/10.1016/j.theriogenology.2004.09.059
  11. Kawakami M, Tani T, Yabuuchi A, Kobayashi T, Murakami H, Fujimura T, Kato Y and Tsunoda Y. 2003. Effect of demecolcine and nocodazole on the efficiency of chemically assisted removal of chromosomes and the developmental potential of nuclear transferred porcine oocytes. Cloning Stem Cells, 5:379-387 https://doi.org/10.1089/153623003772032871
  12. Kidson A, Schoevers E, Langendijk P, Verheijden J, Colenbrander B and Bevers M. 2003. The effect of oviductal epithelial cell co-culture during in vitro maturation on sow oocyte morphology, fertilization and embryo development. Theriogenology, 59:1889-1903 https://doi.org/10.1016/S0093-691X(02)01291-8
  13. Kono T, Kwon OY and Nakahara T. 1991. Development of enucleated mouse oocytes reconstituted with embryonic nu- clei. J. Reprod. Fertil., 93:165-172 https://doi.org/10.1530/jrf.0.0930165
  14. Kurome M, Fujimura T, Murakami H, Takahagi Y, Wako N, Ochiai T, Miyazaki K and Nagashima H. 2003. Comparison of electro-fusion and intracytoplasmic nuclear injection methods in pig cloning. Cloning Stem Cells, 5:367-378 https://doi.org/10.1089/153623003772032862
  15. Magnusson C. 1980. Role of cumulus cells for rat oocyte maturation and metabolism. Gamete Res., 3:133-140 https://doi.org/10.1002/mrd.1120030205
  16. Miao Y, Ma S, Liu X, Miao D, Chang Z, Luo M and Tan J. 2004. Fate of the first polar bodies in mouse oocytes. Mol. Reprod. Dev., 69:66-76 https://doi.org/10.1002/mrd.20148
  17. Miyoshi K, Rzucidlo SJ, Pratt SL and Stice SL. 2002. Utility of rapidly matured oocytes as recipients for production of cloned embryos from somatic cells in the pig. Biol, Reprod., 67:540-545 https://doi.org/10.1095/biolreprod67.2.540
  18. Miyoshi K, Sato K and Yoshida M. 2006. In vitro development of cloned embryos derived from miniature pig somatic cells after activation by ultrasound stimulation. Cloning Stem Cells, 8:159-165 https://doi.org/10.1089/clo.2006.8.159
  19. Mori T, Amano T and Shimizu H. 2000. Roles of gap junctional communication of cumulus cells in cytoplasmic maturation of porcine oocytes cultured in vitro. Biol. Reprod., 62:913-919 https://doi.org/10.1095/biolreprod62.4.913
  20. Motlik J, Fulka J and Flechon JE. 1986. Changes in intercellular coupling between pig oocytes and cumulus cells during maturation in vivo and in vitro. J. Reprod. Fertil., 76:31-37 https://doi.org/10.1530/jrf.0.0760031
  21. Park Y, Hong J, Yong H, Lim J and Lee E. 2005. Effect of exogenous carbohydrates in a serum-free culture medium on the development of in vitro matured and fertilized porcine embryos. Zygote, 13:269-275 https://doi.org/10.1017/S0967199405003369
  22. Petters RM and Wells KD. 1993. Culture of pig embryos. J. Reprod. Fertil. Suppl., 48:61-73
  23. Peura TT, Lewis IM and Trounson AO. 1998. The effect of recipient oocytes volume on nuclear transfer in cattle. Mol. Reprod. Dev., 50:185-191 https://doi.org/10.1002/(SICI)1098-2795(199806)50:2<185::AID-MRD9>3.0.CO;2-G
  24. Polejaeva IA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, Dai Y, Boone J, Walker S, Ayares DL, Colman A and Campbell KH. 2000. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature, 407:86-90 https://doi.org/10.1038/35024082
  25. Talbot P and Dandekar P. 2003. Perivitelline space: dose it play a role in blocking polyspermy in mammals? Microsc. Res. Tech., 61:349-357 https://doi.org/10.1002/jemt.10348
  26. Van de Velde H, De Vos A, Joris H, Nagy ZP and Van Steirteghem AC. 1998. Effect of timing of oocytes denudation and micro-injection on survival, fertilization and em-bryo quality after intracytoplasmic sperm injection. Human Reprod., 13:3160-3164 https://doi.org/10.1093/humrep/13.11.3160
  27. Van der Westerlaken LAJ, Van der Schans A, Eyestone WH and De Boer HA. 1994. Kinetics of first polar body extrusion and the effect of time of stripping of the cumulus and time of insemination on developmental competence of bovine oocytes. Theriogenology, 42:361-370 https://doi.org/10.1016/0093-691X(94)90281-X
  28. Wakayama T, Perry AC, Zuccotti M, Johnson KR and Yanagimachi R. 1998. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature, 394:369-374 https://doi.org/10.1038/28615
  29. Walker SC, Shin T, Zaunbrecher GM, Romano JE, Johnson GA, Bazer FW and Piedrahita JA. 2002. A highly efficient method for porcine cloning by nuclear transfer using in vitromatured oocytes. Cloning Stem Cells, 4:105-112 https://doi.org/10.1089/153623002320253283
  30. Wang W, Abeydeera LR, Prather RS and Day BN. 1998. Morphologic comparison of ovulated and in vitro-matured porcine oocytes, with particular reference to polyspermy after in vitro fertilization. Mol. Reprod. Dev., 49:308-316 https://doi.org/10.1002/(SICI)1098-2795(199803)49:3<308::AID-MRD11>3.0.CO;2-S
  31. Westhusin ME, Collas P, Marek D, Sullivan E, Stepp P, Pryor J and Barnes F. 1996. Reducing the amount of cytoplasm available for early embryonic development decreases the quality but not quantity of embryos produced by in vitro fertilization and nuclear transplantation. Theriogenology, 46:243-252 https://doi.org/10.1016/0093-691X(96)00181-1
  32. Willadsen SM. 1986. Nuclear transplantation in sheep embryos. Nature, 320:63-65 https://doi.org/10.1038/320063a0
  33. Wilmut I, Schnieke AE, Me Whir J, Kind AJ and Campbell KHS. 1997. Viable offspring derived from fetal and adult mammalian cells. Nature, 385:810-813 https://doi.org/10.1038/385810a0
  34. Wongsrikeao P, Kaneshige Y, Ooki R, Taniguchi M, Agung B, Nii M and Otoi T. 2005. Effect of the removal of cumulus cells on the nuclear maturation, fertilization and development of porcine oocytes. Reprod. Dom. Anim., 40:166-170 https://doi.org/10.1111/j.1439-0531.2005.00576.x
  35. Yamauchi N and Nagai T. 1999. Male pronuclear formation in denuded porcine oocytes after in vitro maturation in the presence of cysteamine. Biol. Reprod., 61 :828-833 https://doi.org/10.1095/biolreprod61.3.828