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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Consequences of exposure to serum, with or without vitamin E supplementation, in terms of the fatty acid content and viability of bovine blastocysts produced in vitro

A. Reis A B , J. A. Rooke A , G. J. McCallum A , M. E. Staines A , M. Ewen A , M. A. Lomax B and T. G. McEvoy A C
+ Author Affiliations
- Author Affiliations

A Scottish Agricultural College, Animal Biology Division, Craibstone Estate, Bucksburn, Aberdeen AB21 9YA, UK.

B University of Aberdeen, Department of Agriculture and Forestry, King Street, Aberdeen AB24 5UA, UK.

C To whom correspondence should be addressed. email: t.mcevoy@ab.sac.ac.uk

Reproduction, Fertility and Development 15(5) 275-284 https://doi.org/10.1071/RD03004
Submitted: 8 January 2003  Accepted: 2 July 2003   Published: 7 April 2003

Abstract

To determine whether serum supplementation influenced fatty acid content of bovine blastocysts and whether vitamin E addition to culture medium containing serum could improve development in vitro, cleaved eggs were cultured in synthetic oviduct fluid supplemented with bovine serum albumin (BSA, 0.4% w/v, fraction V) (SVBSA), fetal calf serum (FCS, 10% v/v) (SFCS) or FCS (10% v/v) plus 100 μM vitamin E (SFCS + E). Blastocyst yields were recorded and fatty acid composition was determined by gas chromatography. Day 7 blastocysts were incubated with [2-14C] pyruvate for 3 h and then fixed for cell counts. Yields of good quality blastocysts were greatest from cleaved eggs cultured in serum-free conditions (P < 0.01). In the presence of serum, supplementation with vitamin E increased both total and good quality blastocyst yields (P < 0.01). Presence of serum increased fatty acid content (mean ± SEM) of blastocysts (SVBSA v. SFCS = 57 ± 2  v. 74 ± 2 ng embryo−1; P < 0.001). In contrast, pyruvate metabolism was greater in blastocysts produced without serum (27 ± 3 v. 21 ± 3 picomoles embryo−13 h−1; P < 0.01) but, on a per cell basis, no differences were detected. Addition of vitamin E to the serum-supplemented formulation did not alter either the fatty acid content (73 ± 2 ng embryo−1) or pyruvate metabolism index (19 ± 1 pmol embryo−13 h−1) of SFCS + E blastocysts. Thus, despite lipid accumulation, supplementary vitamin E improved blastocyst yields in embryos exposed to serum.

Extra keywords: antioxidant


Acknowledgments

The Scottish Agricultural College receives funding from the Scottish Executive Environment and Rural Affairs Department. A. Reis is supported by the Portuguese Ministry of Science and Technology (Praxis XXI).


References

Abe, H. , Yamashita, S. , Itoh, T. , Satoh, T. , and Hoshi, H. (1999a). Ultrastructure of bovine embryos developed from in vitro-matured and fertilised oocytes: comparative morphological evaluation of embryos cultured either in serum-free medium or in serum-supplemented medium. Mol. Reprod. Dev. 53, 325–35.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Abe, H. , Otoi, T. , Tachikawa, S. , Yamashita, S. , Satoh, T. , and Hoshi, H. (1999b). Fine structure of bovine morulae and blastocysts in vivo and in vitro.  Anat. Embryol. 199, 519–27.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Abe, H. , Yamashita, S. , Satoh, T. , and Hoshi, H. (2002). Accumulation of cytoplasmic lipid droplets in bovine embryos and cryotolerance of embryos developed in different culture systems using serum-free or serum-containing media. Mol. Reprod. Dev. 61, 57–66.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Barnett, D. , and Bavister, B. (1996). What is the relationship between the metabolism of preimplantation embryos and their developmental competence? Mol. Reprod. Dev. 43, 105–33.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Byrne, A. , Southgate, J. , Brison, D. R. , and Leese, H. J. (1999). Analysis of apoptosis in the preimplantation bovine embryo using TUNEL. J.Reprod. Fertil. 117, 97–105.
PubMed |

Christie, W. W. (1982). Isolation of lipids from tissues. In ‘Lipid Analysis: Isolation, Separation, Identification and Structural Analysis of Lipids’, 2nd edn.  pp. 17–25. (Pergamon Press: Oxford, UK.)

Christie, W. W. , Noble, R. C. , and Moore, J. H. (1970). Determination of lipid classes by gas chromatography procedure. Analyst 95, 940–44.


Crosier, A. , Farin, P. W. , Dykstra, M. J. , Alexander, J. E. , and Farin, C.E. (2000). Ultrastructural morphometry of bovine compact morulae produced in vivo or in vitro.  Biol. Reprod. 62, 1459–65.
PubMed |

Crosier, A. , Farin, P. W. , Dykstra, M. J. , Alexander, J. E. , and Farin, C. E. (2001). Ultrastructural morphometry of bovine blastocysts produced in vivo or in vitro.  Biol. Reprod. 64, 1375–1385.
PubMed |

Dorland, M. , Gardner, D. K. , and Trounson, A. O. (1994). Serum in synthetic oviduct fluid causes mitochondrial degeneration in ovine embryos. J. Reprod. Fertil. Abstract Series 13, 25.(Abstract.)


Ducibella, T. (1998). Biochemical and cellular insights into the temporal windonw of normal fertilization. Theriogenology 49, 53–65.
Crossref | GoogleScholarGoogle Scholar | PubMed |

El Mouatassim, S. , Guerin, P. , and Ménézo, Y. (1999). Expression of genes encoding antioxidant enzymes in human and mouse oocytes during the final stages of maturation. Mol. Hum. Reprod. 5, 720–25.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Fariss, M. W. , Nicholls-Grzemski, F. A. , Tirmenstein, M. A. , and Zhang, J.-G. (2001). Enhanced antioxidant and cryoprotective abilities of vitamin E succinate is associated with a rapid uptake advantage in rat hepatocytes and mitochondria. Free Radical Biol. & Med. 31, 530–41.
Crossref | GoogleScholarGoogle Scholar |

Ferguson, E. M. , and Leese, H. J. (1999). Triglyceride content of bovine oocytes and early embryos. J. Reprod. Fertil. 116, 373–78.
PubMed |

Goodhand, K. L. , Watt, R. G. , Staines, M. E. , Hutchinson, J. S. , and Broadbent, P. J. (1999). In vivo oocyte recovery and in vitro embryo production from bovine donors aspirated at different frequencies or following FSH treatment. Theriogenology 51, 951–61.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Guerin, P. , El Mouatassim, S. , and Ménézo, Y. (2001). Oxidative stress and protection against reactive oxygen species in the preimplantation embryo and its surroundings. Hum. Reprod. Update 7, 175–89.
PubMed |

Harvey, A. J. , Kind, K. L. , and Thompson, J. (2002). REDOX regulation of early embryo development. Biol Reprod. 123, 479–86.


Harvey, M. B. , Arcellana-Panlilio, M. Y. , Zhang, X. , Schultz, G. A. , and Watson, A. J. (1995). Expression of genes encoding antioxidant enzymes in preimplantation mouse and cow embryos and primary bovine oviduct cultures employed for embryo coculture. Biol. Reprod. 53, 532–40.
PubMed |

Homa, S. , Webster, S. , and Russell, R. (1991). Phospholipid turnover and ultrastructural correlates during spontaneous germinal vesicle breakdown of the bovine oocyte: effects of a cyclic AMP phosphodiesterase inhibitor. Dev. Biol. 146, 461–72.
PubMed |

Johnson, M. , and Nasr-Esfahani, M. (1994). Radical solutions and cultural problems: could free oxygen radicals be responsible for the impaired development of preimplantation mammalian embryos in vitro? Bioessays 16, 31–38.
PubMed |

Kamal-Eldin, A. , and Appelqvist, L.-A. (1996). The chemistry and antioxidant properties of tocopherols and tocotrienols. Lipids 31, 671–701.
PubMed |

Kane, M. , Norris, M. , and Harrison, R. (1992). Uptake and incorporation of inositol by preimplantation mouse embryos. J. Reprod. Fertil. 96, 617–25.
PubMed |

Khandoker, M. , and Tsujii, H. (1998). Metabolism of exogenous fatty acids by preimplantation rabbit embryos. J J. Fertil. Steril. 43, 195–201.


Kim, J. , Kinoshita, M. , Ohnishi, M. , and Fukui, Y. (2001). Lipid and fatty acid analysis of fresh and frozen-thawed immature and in vitro matured bovine oocytes. Reproduction 122, 131–8.
PubMed |

Kowaltowski, A. J. , and Vercesi, A. E. (1999). Mitochondrial damage induced by conditions of oxidative stress. Free Radical Biol. & Med. 26, 463–71.
Crossref | GoogleScholarGoogle Scholar |

Kuran, M. , Robinson, J. J. , Staines, M. E. , and McEvoy, T. G. (2001). Development and de novo protein synthetic activity of bovine embryos produced in vitro in different culture systems. Theriogenology 55, 593–606.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Lequarre, A. S. , Feugang, A. , Malhomme, O. , Donnay, I. , Massip, A. , Dessy, F. , and van Langendonckt, A. (2001). Expression of Cu/Zn and Mn superoxide dismutase during bovine embryo development: influence of in vitro culture. Mol. Reprod. Dev. 58, 45–53.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Liebler, D. C. (1993). The role of metabolism in the antioxidant function of vitamin E. Crit. Rev. Toxicol. 23, 147–69.
PubMed |

Liu, L. , and Keefe, D. L. (2000). Cytoplasm mediates both development and oxidation-induced apoptotic cell death in mouse zygotes. Biol. Reprod. 62, 1828–34.
PubMed |

Liu, L. , Trimarchi, J. , and Keefe, L. (2000). Involvement of mitochondria in oxidative stress-induced cell death in mouse zygotes. Biol. Reprod. 62, 1745–53.
PubMed |

McEvoy, T. G. , Robinson, J. J. , Aitken, R. P. , Findlay, P. A. , and Robertson, I. S. (1997). Dietary excesses of urea influence the viability and metabolism of preimplantation sheep embryos and may affect fetal growth among survivors. Anim. Reprod. Sci. 47, 71–90.
Crossref | GoogleScholarGoogle Scholar | PubMed |

McEvoy, T. , Coull, G. , Broadbent, P. J. , Hutchinson, J. S. M. , and Speake, B. K. (2000a). Fatty acid composition in immature cattle, pig and sheep oocytes with intact zona pellucida. J. Reprod. Fertil. 118, 163–70.
PubMed |

McEvoy, T. G. , Sinclair, K. D. , Young, L. E. , Wilmut, I. , and Robinson, J. J. (2000b). Large offspring syndrome and other consequences of ruminant embryo culture in vitro: relevance to blastocyst culture in human ART. Hum. Fertil. 3, 238–46.


McEvoy, T. G. , Robinson, J. J. , and Sinclair, K. D. (2001). Developmental consequences of embryos and cell manipulation in mice and farm animals. Reproduction 122, 507–18.
PubMed |

Nonogaki, T. , Noda, Y. , Goto, Y. , Kishi, J. , and Mori, T. (1994). Developmental blockage of mouse embryos caused by fatty acids. J. Assist. Reprod. Genet. 11, 482–488.
PubMed |

Olson, S. , and Seidel, G. E. (2000). Culture of in vitro-produced bovine embryos with vitamin E improves development in vitro and after transfer to recipients. Biol. Reprod. 62, 248–52.
PubMed |

Pinyopummintr, T. , and Bavister, B. (1991). In vitro matured/in vitro fertilised bovine oocytes can develop into morulae/blastocysts in chemically defined, protein-free culture media. Biol. Reprod. 45, 736–42.
PubMed |

Pratt, H. (1978). Lipids and transitions in embryos. In ‘Development in Mammals’, Vol. 3. (Ed M. H. Johnson)  pp. 83–129. (North-Holland Publishing Co.: Amsterdam.)

Reis, A. , McCallum, G. J. , Staines, M. E. , Ewen, M. , Lomax, M. A. , Rooke, J. A. , and McEvoy, T. G. (2002). Vitamin E supplementation improves bovine embryo development in vitro.  Reproduction Abstr. Ser. 28, 15–16.


Ricciarelli, R. , Zingg, J.-M. , and Azzi, A. A. (2000). Vitamin E reduces the uptake of oxidised LDL by inhibiting CD36 scavenger receptor expression in cultured aortic smooth muscle cells. Circulation 102, 82–7.
PubMed |

Rizos, D. , Lonergan, P. , Boland, M. P. , Arroyo-Garcia, R. , Pintado, B. , de la Fuente, J. , and Gutiérrez-Adán, A. (2002). Analysis of differential messenger RNA expression between bovine blastocysts produced in different culture systems: implications for blastocyst quality. Biol. Reprod. 66, 589–95.
PubMed |

Sata, R. , Tsujii, H. , Abe, H. , Yamashita, S. , and Hoshi, H. (1999). Fatty acid composition of bovine embryos cultured in serum-free and serum-containing medium during early embryonic development. J.Reprod. Dev. 45, 97–103.
Crossref | GoogleScholarGoogle Scholar |

Shamsuddin, M. , and Rodriguez-Martinez, H. (1994). Fine structure of bovine blastocysts developed either in serum-free medium or in conventional co-culture epithelial cells. J. Vet. Med. 41, 307–16.


Sinclair, K. D. , Young, L. E. , Wilmut, I. , and McEvoy, T. G. (2000). In utero overgrowth in ruminants following embryo culture: lessons from mice and a warning to men. Hum. Reprod. 15, 68–86.


Thompson, J. G. , Sherman, A. N. M. , Aleen, N. W. , McGowan, L. T. , and Tervit, H. R. (1998). Total protein content and protein synthesis within pre-elongation stage bovine embryos. Mol. Reprod. Dev. 50, 139–45.
Crossref | GoogleScholarGoogle Scholar | PubMed |

van Saun, R. J. , Herdt, T. H. , and Stowe, H. D. (1989). Maternal and fetal vitamin E concentrations and selenium-vitamin E interrelationships in dairy cattle. J. Nutr. 119, 1156–64.
PubMed |

Zeron, Y. , Ocheretny, A. , Kedar, O. , Borochov, A. , Sklan, D. , and Arav, A. (2001). Seasonal changes in bovine fertility: relation to developmental competence of oocytes, membrane properties and fatty acid composition of follicles. Reproduction 121, 447–54.
PubMed |