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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Accumulation and distribution of neutral lipid droplets is non-uniform in ovine blastocysts produced in vitro in either the presence or absence of serum

A. Reis A B , G. J. McCallum A and T. G. McEvoy A C
+ Author Affiliations
- Author Affiliations

A Scottish Agricultural College, Sustainable Livestock Systems Group, Craibstone Estate, Bucksburn, Aberdeen AB21 9YA, UK.

B Present address: University of Newcastle, Institute for Cell and Molecular Biosciences, Framlington Place, Newcastle upon Tyne NE2 4HH, UK.

C Corresponding author. Email: tom.mcevoy@sac.ac.uk

Reproduction, Fertility and Development 17(8) 815-823 https://doi.org/10.1071/RD05017
Submitted: 14 February 2005  Accepted: 12 October 2005   Published: 16 December 2005

Abstract

Sheep zygotes were cultured in serum-free or serum-supplemented media to determine effects on blastocyst yields and within-blastocyst abundance and distribution of neutral lipid droplets. Embryos cultured in synthetic oviduct fluid supplemented with bovine serum albumin (0.4% w/v) (SBSA) generated similar blastocyst yields (mean ± s.e.m. = 20% ± 5) to those in synthetic oviduct fluid supplemented with serum (10% v/v) from ewes fed a diet containing 0% (SZFO; 26% ± 2) or 3% fish oil (S3FO; 23% ± 3). SBSA zygotes generated more good-quality blastocysts than their SZFO or S3FO counterparts (P < 0.05). Within-blastocyst abundance of neutral lipid droplets was non-uniform; data were collected from discrete embryo sectors (each = 2700 µm2) representing highest (H), intermediate (I) and lowest (L) densities of accumulation. For all sectors, area (µm2) occupied by lipid droplets in SBSA blastocysts (mean H = 470; I = 370; L = 245) was smaller (P < 0.01) than occupied in others (SBSA : SZFO = 1 : 1.41, 1 : 1.48 and 1 : 1.42; SBSA : S3FO = 1 : 1.36, 1 : 1.30 and 1 : 1.31; data for H, I and L, respectively). Among S3FO blastocysts only, inferior quality was associated with greater lipid abundance. Overall, embryo culture in the presence of serum increased neutral lipid droplet abundance but accumulation was non-uniform.


Acknowledgments

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


References

Abe, H. , and Hoshi, H. (2003). Evaluation of bovine embryos produced in high performance serum-free media. J. Reprod. Dev. 49, 193–203.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Abe, H. , Otoi, T. , Tachikawa, S. , Yamashita, S. , Satoh, T. , and Hoshi, H. (1999a). Fine structure of bovine morulae and blastocysts in vivo and in vitro. Anat. Embryol. (Berl.) 199, 519–527.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Abe, H. , Yamashita, S. , Itoh, T. , Satoh, T. , and Hoshi, H. (1999b). 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, 325–335.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Abe, H. , Yamashita, S. , Satoh, T. , and Hoshi, H. (2002a). 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 |

Abe, H. , Matsuzaki, S. , and Hoshi, H. (2002b). Ultrastructural differences in bovine morulae classified as high and low qualities by morphological evaluation. Theriogenology 57, 1273–1283.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Bruce, J. , and Salter, A. (1996). Metabolic fate of oleic acid, palmitic acid and stearic acid in cultured hamster hepatocytes. Biochem. J. 316, 847–852.
PubMed |

Calder, P. , Yaqoob, P. , Harvey, D. , Watts, A. , and Newsholme, E. (1994). Incorporation of fatty acids by concavalin A-stimulated lymphocytes and the effect on fatty acid composition and membrane fluidity. Biochem. J. 300, 509–518.
PubMed |

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, 1459–1465.
Crossref | GoogleScholarGoogle Scholar | 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.
Crossref | GoogleScholarGoogle Scholar | PubMed |

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


Ferguson, E. M. , and Leese, H. J. (1999). Triglyceride content of bovine oocytes and early embryos. J. Reprod. Fertil. 116, 373–378.
PubMed |

Finstad, H. S. , Drevon, C. A. , Kulseth, M. A. , Synstad, A. V. , Knudsen, E. , and Kolset, S. O. (1998). Cell proliferation, apoptosis and accumulation of lipid droplets in U937-1 cells incubated with eicosapentaenoic acid. Biochem. J. 336, 451–459.
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 of following FSH treatment. Theriogenology 51, 951–961.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Kellner-Weibel, G. , McHendry-Rinde, B. , Haynes, M. P. , and Adelman, S. (2001). Evidence that newly synthesised esterified cholesterol is deposited in existing cytoplasmic lipid inclusions. J. Lipid Res. 42, 768–777.
PubMed |

Kim, J. Y. , Kinoshita, M. , Ohnishi, M. , and Kukui, Y. (2001). Lipid and fatty acid analysis of fresh and frozen-thawed immature and in vitro matured bovine oocytes. Reproduction 122, 131–138.
Crossref | GoogleScholarGoogle Scholar | PubMed |

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 |

Lindner, G. M. , and Wright, R. W. (1983). Bovine embryo morphology and evaluation. Theriogenology 20, 407–416.
Crossref | GoogleScholarGoogle Scholar |

McEvoy, T. G. , Coull, G. D. , Broadbent, P. J. , Hutchinson, J. S. , and Speake, B. K. (2000). Fatty acid composition in immature cattle, pig and sheep oocytes with intact zona pellucida. J. Reprod. Fertil. 118, 163–170.
Crossref | GoogleScholarGoogle Scholar | PubMed |

McEvoy, T. G. , Robinson, J. J. , and Sinclair, K. D. (2001). Developmental consequences of embryo and cell manipulation in mice and farm animals. Reproduction 122, 507–518.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Mitchell, L. M. , Reis, A. , Staines, M. E. , Rooke, J. A. , Dingwall, W. S. , and McEvoy, T. G. (2002). Laparoscopic oocyte aspiration from ewes fed diets with or without 3% fish oil at contrasting stages of the breeding season. Theriogenology 57, 614..


Murphy, D. J. , and Vance, J. (1999). Mechanisms of lipid-body formation. Trends Biochem. Sci. 24, 109–115.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Olson, S. E. , and Seidel, G. E. (2000). Culture of in vitro-produced bovine embryos with vitamin E improves development in vitro after transfer to recipients. Biol. Reprod. 62, 248–252.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Onal, A. G. , Robinson, J. J. , Kuran, M. , King, M. E. , Staines, M. E. , Hutchinson, J. S. M. , and McEvoy, T. G. (2000). Effect of dietary supplementation of fish oil on the yield, quality and in vitro metabolism of embryos from superovulated sheep. Theriogenology 53, 281.


Onal, A. G. , Staines, M. E. , Speake, B. K. , Robinson, J. J. , Hutchinson, J. S. M. , and McEvoy, T. G. (2001). Ovine embryo production in vitro using sera with contrasting lipid profiles. Theriogenology 55, 340.


Plante, L. , and King, W. A. (1994). Light and electron microscopic analysis of bovine embryos derived by in vitro and in vivo fertilisation. J. Asst. Reprod. Genet. 11, 515–529.
Crossref | GoogleScholarGoogle Scholar |

Reis, A. , Staines, M. E. , McCallum, G. J. , Rooke, J. A. , Lomax, M. A. , and McEvoy, T. G. (2002). Vitamin E supplementation and a low oxygen atmosphere improve ovine embryo development in a serum-supplemented culture medium. Reprod. Abstr. Ser. 29, 9.


Reis, A. , Rooke, J. A. , McCallum, G. J. , Staines, M. E. , Ewen, M. E. , Lomax, M. A. , and McEvoy, T. G. (2003a). 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. Reprod. Fertil. Dev. 15, 275–284.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Reis, A. , McCallum, G. J. , Staines, M. E. , Ewen, M. E. , Rooke, J. A. , Lomax, M. A. , and McEvoy, T. G. (2003b). Antioxidant provision sustains ovine preimplantation embryo development and mitochondrial function in vitro in the presence of polyunsaturated fatty acid-enriched serum. Pediatr. Res. 53(Suppl., Part 2), 48A.
Crossref | GoogleScholarGoogle Scholar |

Reis, A. , Rooke, J. A. , McCallum, G. J. , Ewen, M. E. , Lomax, M. A. , and McEvoy, T. G. (2003c). Stress-associated developmental arrest following in vitro ovine embryo culture in the presence of docosahexaenoic acid. Reprod. Abstr. Ser. 30, 48.


Rioux, V. , Lemarchal, P. , and Legrand, P. (2000). Myristic acid, unlike palmitic acid, is rapidly metabolized in cultured rat hepatocytes. J. Nutr. Biochem. 11, 198–207.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Rizos, D. , Fair, T. , Papadopoulos, S. , Boland, M. P. , and Lonergan, P. (2002). Developmental, qualitative and ultrastructural differences between ovine and bovine embryos produced in vivo or in vitro. Mol. Reprod. Dev. 62, 320–327.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Rizos, D. , Gutierrez-Adan, A. , Perez-Garnelo, S. , de la Fuente, J. , Boland, M. P. , and Lonergan, P. (2003). Bovine embryo culture in the presence or absence of serum: implications for blastocyst development, cryotolerance, and messenger RNA expression. Biol. Reprod. 68, 236–243.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Rizos, D. , Gutierrez-Adan, A. , Moreira, P. , O’Meara, C. , Fair, T. , Evans, A. C. , Boland, M. P. , and Lonergan, P. (2004). Species-related differences in blastocyst quality are associated with differences in relative mRNA transcription. Mol. Reprod. Dev. 69, 381–386.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Samuel, D. , Paris, S. , and Ailhaud, G. (1976). Uptake and metabolism of fatty acids and analogues by cultured cardiac cells from chick embryo. Eur. J. Biochem. 64, 583–595.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Sata, R. , Tsujii, H. , Abe, H. , Yamashita, S. , and Hochi, S. (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, 307–316.


Tesoriero, J. V. (1981). Early ultrastructural changes of developing oocytes in the dog. J. Morphol. 168, 171–179.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Thompson, J. G. , Gardner, D. K. , Pugh, P. A. , McMillan, W. H. , and Tervit, H. R. (1995). Lamb birth weight is affected by culture system utilized during in vitro pre-elongation development of ovine embryos. Biol. Reprod. 53, 1385–1391.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Tominaga, K. , Hamada, Y. , Yabuue, T. , and Ariyoshi, T. (2000). Effect of linoleic acid-albumin on post-thaw survival of in vitro-produced bovine embryos at the 16-cell stage. J. Vet. Med. Sci. 62, 465–467.
Crossref | GoogleScholarGoogle Scholar | PubMed |

Zeron, Y. , Sklan, D. , and Arav, A. (2002). Effect of polyunsaturated fatty acid supplementation on biophysical parameters and chilling sensitivity of ewe oocytes. Mol. Reprod. Dev. 61, 271–278.
Crossref | GoogleScholarGoogle Scholar | PubMed |