Reproductive and Developmental Biology

  • 제28권4호
  • /
  • Pages.241-245
  • /
  • 2004
  • /
  • 1738-2432(pISSN)
  • /
  • 2288-0151(eISSN)
한국동물번식학회 (The Korean Society of Animal Reproduction)
권호 표지

Nuclear Remodeling and In Vitro Development Following Somatic Cell Nuclear Transfer in Swine

  • Yoon Jong-Taek (Department of Animal Life and Resources, Hankyong National University, Institute of Genetic Engineering, Hankyong National University) ;
  • Kim Yong-Yeup (Kongju National University) ;
  • Lee Jong-Wan (Kongju National University) ;
  • Min Kwan-Sil (The Graduate School of Bio- & Information Technology, Hankyong National University) ;
  • Hwang Seongsoo (Institute of Genetic Engineering, Hankyong National University)
  • 발행 : 2004.12.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

This study was conducted to investigate nuclear remodeling and developmental rate following nuclear transfer of fetal fibroblast cells, ear skin cells and oviduct epithelial cells into porcine recipient oocytes. To test par-thenogenetic activation, oocytes were treated with a 6-dimethylaminopurine (6-DMAP), a single DC-pulse (DC), calcium ionomycin (ionomycin), DC+6-DMAP and ionomycin + 6-DMAP after in vitro maturation. For nuclear transfer, in vitro matured oocytes were enucleated, and donor cells were transferred into oocytes. Cloned embryos were fused and stimulated with 6-DMAP for 4 h and cultured in vitro for 6 days. Among treatments for parthenogenesis, the activation rate of DC +6-DMAP treatment was significantly higher than that of single treatment roups (p<0.01), except for DC treatment group. However, the difference was not significant in activation rate compared to other complex treatment groups. Nuclear swelling of the cloned embryos was initiated at 60 min after stimulation and increased afterwards. Fusion rates were not different among different donor cells. Cleavage rates of DC treatment groups were significantly higher than those of DC+6-DMAP treatment groups (p<0.05) in case that fetal fibroblast and ear cells were used for nuclear donor. The cloned embryos from developed to blastocysts in oviduct epithelial cell nuclear transfer with DC+6-DMAP treatment was significantly higher compared to those with DC only treatment (p<0.05). However, no blastocyst was developed from nuclear transfer of fetal fibroblast and ear cells regardless of activation treatments. Based on these results, a proper activation stimulation may be necessary to increase the activation rate and the development to blastocyst in cloned porcine embryos.

키워드

참고문헌

  1. Beux GL, Richard FJ, Sirard M-A (2003): Effect of cycloheximide, 6-DMAP, roscovitine and butyrolactone I on resumption of meiosis in porcine oocytes. Theriogenology 60:1049-1058
  2. Bordignon V, Smith LC (1998): Telophase enucleation: an improved method to prepare recipient cytoplasts for use in bovine nuclear transfer. Mol Reprod Dev 49:29-36 https://doi.org/10.1002/(SICI)1098-2795(199801)49:1<29::AID-MRD4>3.0.CO;2-Q
  3. Campbell KHS, Loi P, Otaegui PJ, Wilmut I (1996): Cell cycle co-ordination in embryo cloning by nuclear transfer. Rev Reprod 1:4040-4045
  4. Campbell KHS, Richie WA, Wilmut I (1993): Nuclearcytoplasmic interactions during the first cell cycle of nuclear transfer reconstructed bovine embryos: Implications for deoxyribonucleic acid replication and development. Biol Reprod 49:933-942 https://doi.org/10.1095/biolreprod49.5.933
  5. Cheong HT, Takahashi Y, Kanagawa H (1994): Relationship between nuclear remodeling and subsequent development of mouse embryonic nuclei transferred to enucleated oocytes. Mol Reprod Dev 37:138-145 https://doi.org/10.1002/mrd.1080370204
  6. Collas P, Robl JM (1996): Relationship between nuclear remodeling and development in nuclear transplant rabbit embryos. Biol Reprod 45:455-465
  7. Collas P, Balise JJ, Hofmann GA, Robl JM (1992): Influence of cell cycle stage of the donor nuclear on development of nuclear transplant rabbit embryos. Biol Reprod 46:492-500 https://doi.org/10.1095/biolreprod46.3.492
  8. Hagen DR, Prather RS, Forst NL (1991): Response of porcine to electrical and chemical activation during maturation in vitro. Mol Reprod Dev 28:70-73 https://doi.org/10.1002/mrd.1080280111
  9. Hoshi K, Yanagida K, Sato A (1992): Pretreatment of hamster oocytes with Ca-ionophore to facilitate fertilization by ooplasmic intracytoplasmic sperm injection. Hum Reprod 7:871-875 https://doi.org/10.1093/oxfordjournals.humrep.a137752
  10. Jessus C, Rime H, Haccard O, Van Lint J, Goris J, Merlevede W (1991): Tyrosine phosphorylation of $p34^{cdc2}$ and p42 during meiotic maturation of Xenopus oocyte. Antagonistic action of okadaic acid and 6- DMAP. Development 111:813-820
  11. Miyazaki S (1991): Repetitive calcium transients in hamster oocytes. Cell Calcium 12:205-216. https://doi.org/10.1016/0143-4160(91)90021-6
  12. Parrish JJ, Kim CI, Bae IH (1992): Current concepts of cell-cycle regulation and its relationship to oocyte maturation, fertilization and embryo development. Theriogenology 38:277-296 https://doi.org/10.1016/0093-691X(92)90236-K
  13. Prather RS, Eichen PA, Nicks DK, Peters MS (1991): Artificial activation of porcine oocytes matured in vitro. Mol Reprod Dev 28:405-409 https://doi.org/10.1002/mrd.1080280413
  14. Szollosi D, Czolowska, Szollosi, MS, Tarkowski AK (1988): Remodeling of mouse thymocyte nuclei depends on the time on their transfer into activated, homologous oocytes. J Cell Sci 91:603-613
  15. Tsunoda Y, Kato Y (2000): The recent progress on nuclear transfer in mammals. Zool Sci 17:1177-1184
  16. Wang WH, Abeydeera L, Randall R, Prather S, Day BN (1998): Functional analysis of activation of porcine oocytes by spermatozoa, calcium ionophore and electrical pulse. Mol Reprod Dev 51:346-353 https://doi.org/10.1002/(SICI)1098-2795(199811)51:3<346::AID-MRD15>3.0.CO;2-0
  17. Whitaker M, Patel R (1990): Calcium and cell cycle control. Development 108:525-542
  18. Willadsen SM (1986): Nuclear transplantation in sheep embryos. Nature 320: 63-65
  19. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KHS (1997): Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813 https://doi.org/10.1038/385810a0