Czech J. Anim. Sci., 2004, 49(3):93-98 | DOI: 10.17221/4285-CJAS

In vitro aging of porcine oocytes

I. Petrová, M. Sedmíková, E. Chmelíková, D. ©vestková, R. Rajmon
1 , M. S 2, E. C 2, D. © 2, R. R 2 1Grammar School, Říčany near Prague, Czech Republic
2 Department of Veterinary Science, Faculty of Agronomy, Czech University of Agriculture, Prague, Czech Republic

Porcine oocytes matured in vitro develop in various ways if they are further cultivated. In our studies these oocytes were cultivated for 1 to 5 days (in vitro aging). During the 1st day of aging, most of them remained at the stage of metaphase II (98%). Then many oocytes underwent the spontaneous parthenogenetic activation. The portion of activated oocytes reached its peak after 2 or 3 days of aging in vitro (39 or 45%). The portion of fragmented oocytes peaked at the same time (28%). During subsequent aging in vitro (i.e. day 4 or 5 of aging), the portion of lysed oocytes significantly increased (30 or 37%). The highest portion of spontaneously activated parthenogenetic embryos at a pronuclear stage (35%) was observed during the 2nd day of aging in vitro. These pronuclear embryos had mainly one polar body with two pronuclei (47% of all pronuclear embryos) or two polar bodies with one pronucleus (38% of all pronuclear embryos). During the 3rd and 5th day of in vitro aging, there was a significant increase in the portion of parthenogenetic embryos cleaved to the 2-cell or 3-cell stage. When considering the prolonged in vitro culture of porcine oocyte, only the first day of aging should be taken into account, since beyond this time significant changes, i.e. parthenogenesis, fragmentation or lysis, occurred in oocytes under in vitro conditions.

Keywords: pig; oocyte; parthenogenesis; fragmentation

Published: March 31, 2004  Show citation

ACS AIP APA ASA Harvard Chicago Chicago Notes IEEE ISO690 MLA NLM Turabian Vancouver
Petrová I, Sedmíková M, Chmelíková E, ©vestková D, Rajmon R. In vitro aging of porcine oocytes. Czech J. Anim. Sci. 2004;49(3):93-98. doi: 10.17221/4285-CJAS.
Download citation

References

  1. Chian R.C., Nakahara H., Niwa K., Funahashi H. (1992): Fertilization and early cleavage in vitro of aging bovine oocytes a er maturation in culture. Theriogenology, 37, 665-672. Go to original source... Go to PubMed...
  2. Coy P., Romar R. (2002): In vitro production of pig embryos: a point of view. Repris. Fertil. Dev., 14, 275-286. Go to original source... Go to PubMed...
  3. Gable T.L., Woods G.L. (2001): Increasing culture time from 48 to 96 or 144 hours increased the proportion of equine cumulus oocyte complexes with negative or fragmented nucleus morphology. Theriogenology, 55, 1549-1560. Go to original source... Go to PubMed...
  4. Jílek F., Hü elová R., Petr J., Holubová M., Rozinek J. (2000): Activation of pig oocytes using calcium ionophore: effect of protein synthesis inhibitor cycloheximide. Anim. Reprod. Sci., 63, 101-111. Go to original source... Go to PubMed...
  5. Jílek F., Hü elová R., Petr J., Holubová M., Rozinek J. (2001): Activation of pig oocytes using calcium ionophore: effect of the protein kinase inhibitor 6-dimethyl aminopurine. Reprod. Dom. Anim., 36, 139-145. Go to original source...
  6. Kikuchi K., Naito K., Noguchi J., Shimada A., Kaneko H., Yamashita M., Tojo H., Toyoda Y. (1999): Inactivation of p34cdc2 kinase by accumulation of its phosphorylated forms in porcine oocytes matured and aged in vitro. Zygote, 7, 173-179. Go to original source... Go to PubMed...
  7. Kikuchi K., Naito K., Noguchi J., Shimada A., Kaneko H., Yamashita M., Aoki F., Tojo H., Toyoda Y. (2000): Maturation/M-phase promoting factor: A regulator of aging in porcine oocytes. Biol. Reprod., 63, 715-722. Go to original source... Go to PubMed...
  8. Lonergan P., Fair T., Khatir H., Cesaroni G., Mermillod F. (1998): Effect of protein synthesis inhibition before or during in vitro maturation on subsequent development of bovine oocytes. Theriogenology, 50, 417-431. Go to original source... Go to PubMed...
  9. Mailhaes J.B., Young D., London S.N. (1998): Postovulatory ageing of mouse oocytes in vivo and premature centromere separation and aneuploidy. Biol. Reprod., 58, 1206-1210. Go to original source... Go to PubMed...
  10. Mermillod P., Tomanek M., Marchal R., Meijer L. (2000): High developmental competence of ca le oocytes maintained at the germinal vesicle stage for 24 hours in culture by specific inhibition of MPF kinase activity. Mol. Reprod. Dev., 55, 89-95. Go to original source... Go to PubMed...
  11. Motlík J., Fulka J. (1976): Breakdown of germinal vesicle in pig oocytes in vivo and in vitro. J. Exp. Zool., 198, 155-162. Go to original source... Go to PubMed...
  12. Motlík J., Kubelka M. (1990): Cell-cycle aspects of growth and maturation of mammalian oocytes. Mol. Reprod. Dev., 27, 366-375. Go to original source... Go to PubMed...
  13. Orrenius S., Zhivotovsky B., Nicotera P. (2003): Regulation of cell death: The calcium-apoptosis link. Nature Rev. Molec. Cell. Biol., 4, 552-565. Go to original source... Go to PubMed...
  14. Pavlok A., Kaláb P., Bobák P. (1997): Fertilisation competence of bovine normally matured or aged oocytes derived from different antral follicles: morphology, protein synthesis, H1 and MBP kinase activity. Zygote, 5, 235-246. Go to original source... Go to PubMed...
  15. Perez G.I., Tilly J.L. (1997): Cumulus cells are required for the increased apoptotic potential in oocytes of aged mouse. Human Reprod., 12, 2781-2783. Go to original source... Go to PubMed...
  16. Perez G.I., Tao X.J., Tilly J.L. (1999): Fragmentation and death (a.k.a. apoptosis) of ovulated oocytes. Molec. Human Reprod., 5, 414-420. Go to original source... Go to PubMed...
  17. Petr J., Grocholová R., Rozinek J., Jílek F. (1995): Activation of in vitro matured pig oocytes by combined treatment of ethanol and cycloheximide. Theriogenology, 45, 1473-1478. Go to original source...
  18. Petr J., Rozinek J., Jílek F., Urbánková D. (2000): Activation of porcine oocytes using cyclopiazonic acid, an inhibitor of calcium-dependent ATPases. J. Exp. Zool., 287, 304-315. Go to original source...
  19. Petr J., Urbánková D., Tománek M., Rozinek J., Jílek F. (2002): Activation of in vitro matured pig oocytes using activators of inositol triphosphate or ryanodine receptors. Anim. Reprod. Sci., 70, 235-249. Go to original source... Go to PubMed...
  20. Pinyopummin A., Takahashi Y., Hishimuna K., Kanagawa H. (1993): Development of haploid and diploid mouse parthenotes - effect of oocyte aging in-vivo. Jap. J. Vet. Res., 41, 81-87.
  21. Snedecor G.W., Cochran W.G. (1957): Statistical methods, Iowa: Iowa State University Press.
  22. Suzuki H., Takashima Y., Toyokawa K. (2002): Cytoskeletal organization pf porcine oocytes aged and activated electrically or by sperm. J. Reprod. Dev., 48, 293-301. Go to original source...
  23. Takahashi T., Saito H., Hiroi M., Doi K., Takahashi E. (2000): Effects of aging on inositol 1,4,5-triphosphateinduced Ca2+ release in unfertilized mouse oocytes. Molec. Reprod. Dev., 55, 299-306. Go to original source...
  24. Takahashi T., Takahashi E., Igarishi H., Tezuka N., Kurachi H. (2003): Impact of oxidative stress in aged oocytes on calcium oscillations at fertilization. Molec. Reprod. Dev., 66, 143-152. Go to original source... Go to PubMed...
  25. Wassarman P.M. (1988): The mammalian ovum. In: Knobil E., Neill J. (eds.): The Physiology of Reproduction. Raven Press, New York. 69-102.
  26. Yanagimachi R. (1988): Mammalian fertilization. In: Knobil E., Neill J. (eds.): The Physiology of Reproduction. Raven Press, New York. 135-185.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International (CC BY NC 4.0), which permits non-comercial use, distribution, and reproduction in any medium, provided the original publication is properly cited. No use, distribution or reproduction is permitted which does not comply with these terms.