ARSci  Vol.2 No.3 , August 2014
The Conditions of Ovary Storage Affect the Quality of Porcine Oocytes
Abstract: We studied the effect of different storage conditions of porcine ovaries (time, temperature) on the characteristics of the follicular fluid, immature oocyte quality, meiotic competence and in vitro fertilization of oocytes. Ovaries were stored for 2, 4 or 6 h at 3 different temperatures (15°C, 25°C or 35°C). As the storage time increased, pH and glucose concentration of the follicular fluid, percentage of live immature oocytes at germinal vesicle stage, oxidative activity and maturation rate decreased. At higher temperatures, pH and glucose concentration decreased, but oxidative activity and oocyte maturation rate increased. Lactate concentration and immature oocyte ROS production increased as storage time and temperature increased. The ovary storage for longer than 2 h at 25°C and 35°C resulted in low pH of the follicular fluid and high ROS level in immature oocytes. Such conditions seem to damage oocytes and impair their meiotic competence. A decrease in the oxidative activity caused by long time and/or low storage temperature may imply a decrease in oocyte vitality. In conclusion, in the porcine species, the transport of ovaries at 25°C and 35°C for 2 h are the best conditions to maintain adequate oocyte quality, meiotic competence and in vitro fertilization rates.
Cite this paper: Tellado, M. , Alvarez, G. , Dalvit, G. and Cetica, P. (2014) The Conditions of Ovary Storage Affect the Quality of Porcine Oocytes. Advances in Reproductive Sciences, 2, 56-67. doi: 10.4236/arsci.2014.23007.

[1]   Guignot, F., Bezard, J. and Palmer, E. (1999) Effect of Time during Transport of Excised Mare Ovaries on Oocyte Recovery Rate and Quality after in Vitro Maturation. Theriogenology, 52, 757-766.

[2]   Moodie, G. and Graham, E. (1989) The Effect of Incubating Sheep Ovaries for Various Times and Temperatures on Oocyte Maturation Rates in Vitro. Biology of Reproduction, 40, 53.

[3]   Evecen, M., Cirit, U., Demir, K., Ozdas, O.B., Tas, M., Birler, S. and Pabuccuoglu, S. (2010) Effects of Estrous Cycle Stage and Transport Temperature of Ovaries on in Vitro Maturation of Canine Oocytes. Animal Reproduction Science, 117, 160-165.

[4]   Yang, N., Lu, K. and Gordon, I. (1990) In Vitro Fertilization (IVF) and Culture (IVC) of Bovine Oocytes from Stored Ovaries. Theriogenology, 33, 352.

[5]   Solano, R., De Armas, R., Pupo, C. and Castro, F. (1994) Short Term Preservation of Intrafollicular Oocytes at 4 C. Theriogenology, 41, 299.

[6]   Evecen, M., Cirit, U., Demir, K., Karaman, E., Hamzaoglu, A.I. and Bakirer, G. (2009) Developmental Competence of Domestic Cat Oocytes from Ovaries Stored at Various Durations at 4°C Temperature. Animal Reproduction Science, 116, 169-172.

[7]   Wongsrikeao, P., Otoi, T., Karja, N.W., Agung, B., Nii, M. and Nagai, T. (2005) Effects of Ovary Storage Time and Temperature on DNA Fragmentation and Development of Porcine Oocytes. Journal of Reproduction and Development, 51, 87-97.

[8]   Lin, Y.A., Tsai, H.B., Liao, M.H. and Chen, M.C. (2010) Effect of Preserving Condition of Porcine Ovaries on the Development of in Vitro Matured Oocytes. Reproduction in Domestic Animals, 46, 333-337.

[9]   Nelson, D. and Cox, M. (2005) Enzymes. In: Nelson, D. and Cox, M., Eds., Lehninger Principles of Biochemistry, 4th Edition, W. H. Freeman and Company, New York, 190-237.

[10]   Chang, S.C., Jones, J.D., Ellefson, R.D. and Ryan, R.J. (1976) The Porcine Ovarian Follicle: I. Selected Chemical Analysis of Follicular Fluid at Different Developmental Stages. Biology of Reproduction, 15, 321-328.

[11]   Orsi, N.M., Gopichandran, N., Leese, H.J., Picton, H.M. and Harris, S.E. (2005) Fluctuations in Bovine Ovarian Follicular Fluid Composition throughout the Oestrous Cycle. Reproduction, 129, 219-228.

[12]   Nelson, D. and Cox, M. (2005) Glycolysis, Gluconeogenesis, and the Pentose Phosphate Pathway. In: Nelson, D. and Cox, M., Eds., Lehninger Principles of Biochemistry, 4th Edition. W. H. Freeman and Company, New York, 521-559.

[13]   Fatehi, A.N., Roelen, B.A., Colenbrander, B., Schoevers, E.J., Gadella, B.M., Beverst, M.M. and van den Hurk, R. (2005) Presence of Cumulus Cells during in Vitro Fertilization Protects the Bovine Oocyte against Oxidative Stress and Improves First Cleavage but Does Not Affect Further Development. Zygote, 13, 177-185.

[14]   Choi, W.J., Banerjee, J., Falcone, T., Bena, J., Agarwal, A. and Sharma, R.K. (2007) Oxidative Stress and Tumor Necrosis Factor-Alpha-Induced Alterations in Metaphase II Mouse Oocyte Spindle Structure. Fertility and Sterility, 88, 1220-1231.

[15]   Tatemoto, H., Sakurai, N. and Muto, N. (2000) Protection of Porcine Oocytes against Apoptotic Cell Death Caused by Oxidative Stress during in Vitro Maturation: Role of Cumulus Cells. Biology of Reproduction, 63, 805-810.

[16]   Zhang, X., Li, X.H., Ma, X., Wang, Z.H., Lu, S. and Guo, Y.L. (2006) Redox-Induced Apoptosis of Human Oocytes in Resting Follicles in Vitro. Journal of the Society for Gynecologic Investigation, 13, 451-458.

[17]   Lopes, A.S., Lane, M. and Thompson, J.G. (2010) Oxygen Consumption and ROS Production Are Increased at the Time of Fertilization and Cell Cleavage in Bovine Zygotes. Human Reproduction, 25, 2762-2773.

[18]   Dumollard, R., Ward, Z., Carroll, J. and Duchen, M.R. (2007) Regulation of Redox Metabolism in the Mouse Oocyte and Embryo. Development, 134, 455-465.

[19]   Cunningham, J.G. and Klein, B.G. (2007) Textbook of Veterinary Physiology. Elsevier Science Health Science Division.

[20]   Barham, D. and Trinder, P. (1972) An Improved Colour Reagent for the Determination of Blood Glucose by the Oxidase System. Analyst, 97, 142-145.

[21]   Gutnisky, C., Dalvit, G.C., Pintos, L.N., Thompson, J.G., Beconi, M.T. and Cetica, P.D. (2007) Influence of Hyaluronic Acid Synthesis and Cumulus Mucification on Bovine Oocyte in Vitro Maturation, Fertilisation and Embryo Development. Reproduction, Fertility and Development, 19, 488-497.

[22]   Alvarez, G.M., Dalvit, G.C., Achi, M.V., Miguez, M.S. and Cetica, P.D. (2009) Immature Oocyte Quality and Maturational Competence of Porcine Cumulus-Oocyte Complexes Subpopulations. Biocell, 33, 167-177.

[23]   Hoppe, R. and Bavister, B. (1984) Evaluation of the Fluorescein Diacetate (FDA) Vital Dye Viability Test with Hamster and Bovine Embryos. Animal Reproduction Science, 6, 323-325.

[24]   Luttmer, S. and Longo, F. (1986) Examination of Living and Fixed Gametes and Early Embryos Stained with Supravital Fluorochromes. Gamete Research, 15, 267-283.

[25]   Abeydeera, L.R., Wang, W.H., Prather, R.S. and Day, B.N. (2001) Effect of Incubation Temperature on in Vitro Maturation of Porcine Oocytes: Nuclear Maturation, Fertilisation and Developmental Competence. Zygote, 9, 331-337.

[26]   Hancock, J. and Hovell, G. (1959) The Collection of Boar Semen. The Veterinary Record, 71, 664-665.

[27]   Abeydeera, L.R. and Day, B.N. (1997) In Vitro Penetration of Pig Oocytes in a Modified Tris-Buffered Medium: Effect of BSA, Caffeine and Calcium. Theriogenology, 48, 537-544.

[28]   Pereira, R.J., Tuli, R.K., Wallenhorst, S. and Holtz, W. (2000) The Effect of Heparin, Caffeine and Calcium Ionophore A23187 on in Vitro Induction of the Acrosome Reaction in Frozen-Thawed Bovine and Caprine Spermatozoa. Theriogenology, 54, 185-192.

[29]   LeBel, C., Ischiropoulos, H. and Bondy, S. (1992) Evaluation of the Probe 2',7'-Dichlorofluorescein as an Indicator of Reactive Oxygen Species Formation and Oxidative Stress. Chemical Research in Toxicology, 5, 227-231.

[30]   Morado, S.A., Cetica, P.D., Beconi, M.T. and Dalvit, G.C. (2009) Reactive Oxygen Species in Bovine Oocyte Maturation in Vitro. Reproduction, Fertility and Development, 21, 608-614.

[31]   Lane, M., Maybach, J.M. and Gardner, D.K. (2002) Addition of Ascorbate during Cryopreservation Stimulates Subsequent Embryo Development. Human Reproduction, 17, 2686-2693.

[32]   Somfai, T., Imai, K., Kaneda, M., Akagi, S., Watanabe, S., Haraguchi, S., Mizutani, E., Dang-Nguyen, T.Q., Inaba, Y., Geshi, M. and Nagai, T. (2011) The Effect of Ovary Storage and in Vitro Maturation on mRNA Levels in Bovine Oocytes; A Possible Impact of Maternal ATP1A1 on Blastocyst Development in Slaughterhouse-Derived Oocytes. Journal of Reproduction and Development, 57, 723-730.

[33]   Oakberg, E.F. (1979) Follicular Growth and Atresia in the Mouse. In Vitro, 15, 41-49.

[34]   Murdoch, W.J. (1992) Comparative Morphometry and Steroidogenic Function of Antral Ovine Follicles Destined for Ovulation or Atresia. Domestic Animal Endocrinology, 9, 219-224.

[35]   Assey, R.J., Hyttel, P., Greve, T. and Purwantara, B. (1994) Oocyte Morphology in Dominant and Subordinate Follicles. Molecular Reproduction and Development, 37, 335-344.

[36]   Cetica, P.D., Dalvit, G.C. and Beconi, M.T. (1999) Study of Evaluation Criteria Used for in Vitro Bovine Oocyte Selection and Maturation. Biocell, 23, 125-133.