Effects of Genotypes on In Vitro Maturation and Fertilization of Frozen-Thawed Porcine Oocytes

  • Jia Y. H. (College of Animal Resource Science, Kangwon National University, College of Animal Science and Technology, Laiyang Agricultural University) ;
  • Jin H. J. (Animal Genetic Resources Station, National Livestock Research Institute) ;
  • Wee M. S. (College of Animal Resource Science, Kangwon National University, Animal Genetic Resources Station, National Livestock Research Institute) ;
  • Cheong H. T. (College of Animal Resource Science, Kangwon National University) ;
  • Yang B. K. (College of Animal Resource Science, Kangwon National University) ;
  • Park C. K. (College of Animal Resource Science, Kangwon National University)
  • Published : 2005.12.01

Abstract

In the present study, we investigated the effects of genotypes on in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes. The porcine cumulus-oocyte complexes (COCs) were divided into four groups according to whether they were: (1) in vitro matured; (2) cryopreserved and in vitro matured; (3) in vitro fertilized and (4) cryopreserved, and in vitro fertilized. Maturation of porcine COCs was accomplished by incubation in NCSU23 medium. Immature oocytes were cryopreserved by Open Pulled Straws (OPS) method according to Vajta et al., (1998). Oocytes stained by Acetic-Orcein method were observed under the microscope. DNA extracted from the ovaries was analyzed by RAPD (random amplified polymorphic DNA) and SSCP (single strand conformational polymorphisrrt) method. The rates of oocytes maturation and fertilization were significantly high in AA genotype. The results indicated that in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes may be affected by genotypes in pigs.

Keywords

References

  1. Arens M (1999): Methods for subtyping and molecular comparison of human viral genomes. Clinical Microbiology Reviews 124:612-626
  2. Baker RD, Polge C (1976): Fertilization in swine and cattle. Canadian J Anim Sci 56: 105-109 https://doi.org/10.4141/cjas76-016
  3. Binder T, Siegert W, Kruse A, Oettle H, Wilborn F, Peng R, Timm H, Neuhaus P, Schmidt CA (1999): Identification of human cytomegalovirus variants by analysis of single-strand conformation polymorphism and DNA sequencing of the envelope glycoprotein B gene region -distribution frequency in liver transplant recipients. Journal of Virological Methods 78:153-162 https://doi.org/10.1016/S0166-0934(98)00173-6
  4. Bjerregaard B, Maddox-Hyttel P (2004): Regulation of ribosomal RNA gene expression in porcine oocytes. Animal Reproduction Science 83:605-616 https://doi.org/10.1016/j.anireprosci.2004.04.023
  5. Cheng WTK (1985): In vitro fertilization of farm animal oocytes. PhD Thesis. Council of National Academic Awards Cambridge. UK
  6. Didion BA, Pomp D, Martin MJ, Homanics GE, Markert CL (1990): Observations on the cooling and cryopreservation of pig oocytes at the germinal vesicle stage. J Anim Sci 68:2803-2810 https://doi.org/10.2527/1990.6892803x
  7. Eroglu A, Toth TL, Toner M (1998): Alterations of the cytoskeleton and polyploidy induced by cryopreservation of metaphase II mouse oocytes. Fertil Steril 69:944-957 https://doi.org/10.1016/S0015-0282(98)00030-2
  8. Fuku E, Xia L, Downey BR (1995): Ultrastructural changes in bovine oocytes cryopreserved by vitrification. Cryobiology 32:139-156 https://doi.org/10.1006/cryo.1995.1013
  9. Hochi S, Fujimoto T, Oguri N (1995): Viability of immature horse oocytes cryopreserved by vitrification. Theriogenology 43:236 https://doi.org/10.1016/0093-691X(95)92390-U
  10. Iritani A, Niwa K. Imai H (1978): Sperm penetration in vitro of pig follicular oocytes matured in culture. J Reprod Fert 54:379-383 https://doi.org/10.1530/jrf.0.0540379
  11. Isachenko V, Soler C, Isachenko E, Perez-Sanchez F, Grishchenko V (1998): Vitrification of immature porcine oocytes: effects of lipid droplets, temperature, cytoskeleton, and addition and removal of cryoprotectant. Cryobiology 36:250-253 https://doi.org/10.1006/cryo.1998.2079
  12. Lim JM, Fukui Y, Ono H (1992): Developmental competence of bovine oocytes frozen at various maturation stages followed by in vitro maturation and fertilization Theriogenology 37:351-361 https://doi.org/10.1016/0093-691X(92)90193-U
  13. Lin JC, Kumar B, Lin SC (1993): Rapid and sensitive genotyping of Epstein-Barr virus using single-strand conformation polymorphism analysis of polymerase chain reaction products. Joumal of Virological Methods 43:233-246 https://doi.org/10.1016/0166-0934(93)90079-7
  14. Liu RH, Sun QY, Li YH, Jiao LH, Wang WH (2003): Effects of cooling on meiotic spindle structure and chromosome alignment within in vitro matured porcine oocytes. Mol Reprod 65:212-218 https://doi.org/10.1002/mrd.10282
  15. Martino A, Songsasen N, Leibo SP (1996): Development into blastocysts of bovine oocytes cryopreserved by ultra-rapid cooling. BioI Reprod 54: 1059-1069 https://doi.org/10.1095/biolreprod54.5.1059
  16. Mattioli M, Bacci ML, Galeati G, Seren E (1989): Developmental competence of pig oocytes matured and fertilized in vitro. Theriogenology 31: 1201-1207 https://doi.org/10.1016/0093-691X(89)90089-7
  17. Motlik J, Fulka J (1974): Fertilization of pig oocytes cultivated in vitro. J Reprod Fertil 36:235-237 https://doi.org/10.1530/jrf.0.0360235
  18. Orita M, Suzuki Y, Sekiya T, Hayashi K (1989): Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 5:874-879 https://doi.org/10.1016/0888-7543(89)90129-8
  19. Otoi T, Yamamoto K. Koyama N, Suzuki T (1995): In vitro fertilization and development of immature and mature bovine oocytes cryopreserved by ethylene glycol with sucrose. Cryobiology 32:455-460 https://doi.org/10.1006/cryo.1995.1045
  20. Palacio A, Duran-Vila N (1999): Single-strand conformation polymorphism (SSCP) analysis as a tool for viroid characterisation. Journal of Virological Methods 77:27-36 https://doi.org/10.1016/S0166-0934(98)00121-9
  21. Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T. (1998): Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol Reprod 12:53-58 https://doi.org/10.1002/(SICI)1098-2795(199809)51:1<53::AID-MRD6>3.0.CO;2-V
  22. Vieira AD, Mezzalira A, Barbieri DP, Lehmkuhl RC, Rubin MI, Vajta G (2002): Calves born after open pulled straw vitrification of immature bovine oocytes. Cryobiology 45:91-94 https://doi.org/10.1016/S0011-2240(02)00109-8
  23. Webb M, Howlett SK. Maro B (1986): Parthenogenesis and cytoskeletal organization in ageing mouse eggs. J Embryol Exp Morphol 95: 131-145
  24. Williams JGK. Kubelik AR, Livak KJ, Rafalski A, Tingey SV (1990): DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research 18:531-535 https://doi.org/10.1093/nar/18.3.531