Hostname: page-component-8448b6f56d-jr42d Total loading time: 0 Render date: 2024-04-18T19:48:59.442Z Has data issue: false hasContentIssue false
  • English
  • Français

Ultrastructural characterization of in vivo-produced ovine morulae and blastocysts

Published online by Cambridge University Press:  30 July 2014

E.M.V. Bettencourt ,
C.M.V. Bettencourt ,
J.N. Chagas e Silva ,
P. Ferreira ,
E. Oliveira ,
R. Romão ,
A. Rocha  and
M. Sousa
E.M.V. Bettencourt*
Affiliation:
Department of Veterinary Medicine, University of Évora, 7000-092 Évora, Portugal. Department of Veterinary Medicine, University of Évora, 7000-092 Évora, Portugal. ICAAM – Instituto de Ciências Agrárias e Ambientais Mediterrâneas. Universidade de Évora, Núcleo da Mitra, Ap. 94 7002–554 Évora, Portugal.
C.M.V. Bettencourt
Affiliation:
Regional Agriculture Direction of Alentejo (DRAAL), Herdade da Abóbada, V.N.S. Bento, Portugal.
J.N. Chagas e Silva
Affiliation:
Veterinary Medicine Faculty, Reproduction Department, Lisbon University (UL), Lisbon, Portugal.
P. Ferreira
Affiliation:
ICBAS, CECA/ICETA, University of Porto, da UP, Portugal.
E. Oliveira
Affiliation:
Department of Microscopy, Laboratory of Cell Biology, UMIB, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Portugal.
R. Romão
Affiliation:
ICAAM – Instituto de Ciências Agrárias e Ambientais Mediterrâneas. Universidade de Évora, Núcleo da Mitra, Ap. 94 7002–554 Évora, Portugal. Department of Zootechny, University of Évora, 7000-092 Évora, Portugal.
A. Rocha
Affiliation:
ICBAS, CECA/ICETA, University of Porto, da UP, Portugal.
M. Sousa
Affiliation:
Department of Microscopy, Laboratory of Cell Biology, UMIB, Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Portugal.
*
All correspondence to: Elisa Maria Varela Bettencourt. Department of Veterinary Medicine, University of Évora, 7000-092 Évora, Portugal. Tel: +35 1266760809. Fax: +35 1266760944. e-mail: emvb@uevora.pt
Get access Rights & Permissions [Opens in a new window]

Summary

The ultrastructure of in vivo-produced ovine embryos, at the morula, early blastocyst and late blastocyst stages, was evaluated using transmission electron microscopy. Embryonic cells were characterized by the presence of intact intercellular junctions, numerous mitochondria, smooth endoplasmic reticulum cisternae and light vesicles. Polyribosomes, rough endoplasmic reticulum cisternae, secondary lysosomes, Golgi complexes and lipid droplets were also observed in the cytoplasm. The nucleus was well defined and organized, with an intact envelope rich in nuclear pore complexes, and one or more reticular nucleoli. Microvilli were present in external blastomeres of morulae and became more abundant in trophectoderm cells of early and late blastocysts. Light vesicles seemed to be associated with small cisternae of Golgi and endoplasmic reticulum origin. These cisternae fused and created light vesicles with engulfed heterogeneous cytosolic structures, small cisternae and vesicles. Their labile membrane enabled them to rapidly coalesce into medium-sized vesicles that began to engulf mitochondria and lipid droplets, forming giant vacuoles mostly filled with fat. Incomplete matured secretory vesicles were observed to exocytose into the perivitelline space of morulae, whereas fully matured secretory vesicles appeared only in trophectoderm cells, being exocytosed into the blastocoelic cavity. These observations suggested that these endoplasmic-/Golgi-derived vesicles behave as active autophagic organelles presenting probably a maturation process from compact morulae to blastocyst.

Keywords

BlastocystMorulaOvineUltrastructureVesicles
Type
Research Article
Information
Zygote , Volume 23 , Issue 4 , August 2015 , pp. 583 - 593
Copyright
Copyright © Cambridge University Press 2014 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Abe, H., Otoi, T., Tachikawa, S., Yamashita, S., Satoh, T. & Hoshi, H. (1999). Fine structure of bovine morulae and blastocyst in vivo and in vitro. Anat. Embryol. 199, 519–27.CrossRefGoogle ScholarPubMed
Abe, H., Matsuzaki, S. & Hoshi, H. (2002). Ultrastructural differences in bovine morulae classified as high and low qualities by morphological evaluation. Theriogenology 57, 1273–83.CrossRefGoogle ScholarPubMed
Bettencourt, E.M.V., Bettencourt, C.M., Silva, J.N.C., Ferreira, P., Matos, C.P., Oliveira, E., Romão, R.J., Rocha, A. & Sousa, M. (2009). Ultrastructural characterization of fresh and cryopreserved in vivo produced ovine embryos. Theriogenology 71, 947–58.CrossRefGoogle ScholarPubMed
Boni, R., Tosti, E., Roviello, S. & Dale, B. (1999). Intracellular communication in in vivo and in vitro produced bovine embryos. Biol. Reprod. 61, 1050–5.CrossRefGoogle Scholar
Calarco, P.G. & McLaren, A. (1976). Ultrastructural observations of preimplantation stages of the sheep. J. Embryol. Exp. Morph. 36, 609–22.Google ScholarPubMed
Carvalho, F., Sousa, M., Oliveira, E., Carvalheiro, J. & Baldaia, L. (1998a). Ultrastructure of oogenesis in Penaeus kerathurus (Crustacea, Decapoda). I Previtellogenic oocytes. J. Sumicrosc. Cytol. Pathol. 30, 408–16.Google ScholarPubMed
Carvalho, F., Sousa, M., Oliveira, E., Carvalheiro, L.& Baldaia, L. (1998b). Ultrastructure of oogenesis in Penaeus kerathurus (Crustacea, Decapoda). II Vitellogenesis. J. Sumicrosc. Cytol. Pathol. 30, 527–35.Google Scholar
Carvalho, F., Sousa, M., Oliveira, E., Carvalheiro, J. & Baldaia, L. (1999). Ultrastructure of oogenesis in Penaeus kerathurus (Crustacea, Decapoda). III Cortical vesicle formation. J. Sumicrosc. Cytol. Pathol. 31, 57–63.Google Scholar
Cocero, M.J., Díaz de la Espina, S.M. & Aguilar, B. (2002). Ultrastructural characteristics of fresh and frozen-thawed ovine embryos using two cryoprotectants. Biol. Reprod. 66, 1244–58.CrossRefGoogle ScholarPubMed
Cognié, Y., Baril, G., Poulin, N. & Mermillod, P. (2003). Current status of embryo technologies in sheep and goats. Theriogenology 59, 171–88.CrossRefGoogle Scholar
Crosier, A.E., Farin, P.W., Dykstra, M.J., Alexander, J.E. & Farin, C.E. (2000). Ultrastructural morphometry of bovine compact morulae produced in vivo or in vitro. Biol. Reprod. 62, 1459–65.CrossRefGoogle ScholarPubMed
Crosier, A.E., Farin, P.W., Dykstra, M.J., Alexander, J.E. & Farin, C.E. (2001). Ultrastructural morphometry of bovine blastocysts produced in vivo or in vitro. Biol. Reprod. 64, 1375–85.CrossRefGoogle ScholarPubMed
Cuello, C., Berthelot, F., Delaleu, B., Venturi, E., Pastor, L.M., Vazquez, J.M., Roca, J., Martinat-Botté, F. & Martinez, E.A. (2007). The effectiveness of the stereomicroscopic evaluation of embryo quality in vitrified-warmed porcine blastocysts: An ultrastructural and cell death study. Theriogenology 67, 970–82.CrossRefGoogle ScholarPubMed
Dobrinsky, J.R. (1996). Cellular approach to cryopreservation of embryos. Theriogenology 45, 17–26.CrossRefGoogle Scholar
Dorland, M., Gardner, D.K. & Trounson, A.O. (1994). Serum in synthetic oviduct fluid causes mitochondrial degeneration in ovine embryos. J. Reprod. Fertil. Abstr. Ser. 13, 25.Google Scholar
Fabian, D., Gjorret, J.O., Berthelot, F., Martinat-Botté, F. & Maddox-Hyttel, P. (2005). Ultrastructure and cell death of in vivo derived and vitrified porcine blastocysts. Mol. Reprod. Dev. 70, 155–65.CrossRefGoogle ScholarPubMed
Fair, T., Lonergan, P., Dinnyes, A., Cottell, D.C., Hyttel, P., Ward, F.A. & Boland, M.P. (2001). Ultrastructure of bovine blastocysts following cryopreservation: effect of method of blastocyst production. Mol. Reprod. Dev. 58, 186–95.3.0.CO;2-N>CrossRefGoogle ScholarPubMed
Ferrer, F., García, C., Villar, J. & Arias, M. (1995). Ultrastructural study of the early development of the sheep embryo. Anat. Histol. Embryol. 24, 191–6.CrossRefGoogle ScholarPubMed
Hyttel, P., Laurincik, J., Viuff, D., Fair, T., Zakhartchenko, V., Rosenkranz, C., Avery, B., Rath, D., Niemann, H. & Thomsen, P.D.et al. (2000). Activation of ribosomal RNA genes in preimplantation cattle and swine embryos. Anim. Reprod. Sci. 60–61, 49–60.CrossRefGoogle ScholarPubMed
Hyttel, P., Viuff, D., Fair, T., Laurincik, J., Thomsen, P.D., Callesen, H., Vos, P.L., Hendriksen, P.J., Dieleman, S.J. & Schellander, K.et al. (2001). Ribosomal RNA gene expression and chromosome aberrations in bovine oocytes and preimplantation embryos. Reproduction 122, 21–30.CrossRefGoogle ScholarPubMed
Linares, T. & Plõen, L. (1981). On the ultrastructure of seven day old normal (blastocysts) and abnormal bovine embryos. Anat. Embryol. 10, 212–26.CrossRefGoogle ScholarPubMed
Linder, G.M. & Wright, R.W. (1983). Bovine embryo morphology and evaluation. Theriogenology 20, 407–16.CrossRefGoogle Scholar
McKelvey, A.C., Robinson, J.J., Aitken, R.P. & Henderson, G. (1985). The evaluation of a laparoscopic insemination technique in ewes. Theriogenology 24, 519–35.CrossRefGoogle ScholarPubMed
Rizos, D., Fair, T., Papadopoulos, S., Boland, M.P. & Lonergan, P. (2002). Developmental, qualitative and ultrastructural differences between ovine and bovine embryos produced in vivo or in vitro. Mol. Reprod. Dev. 62, 320–7.CrossRefGoogle ScholarPubMed
Smith, C.L. & Murphy, C.A. (1987). An antegrade surgical uterine flush technique for ova collection in the ewe. Am. J. Vet. Res. 48, 1129–31.Google ScholarPubMed
Smith, G.D. & Silva, C.A.S.E. (2004). Developmental consequences of cryopreservation of mammalian oocytes and embryos. Reprod. BioMed. Online 9, 171–8.CrossRefGoogle ScholarPubMed
Stringfellow, D.A. & Seidel, S.M. (eds) (1998). Manual of the International Embryo Transfer Society. Savoy, IL, USA: International Embryo Transfer Society, Inc.Google Scholar
Vajta, G., Hyttel, P. & Callesen, H. (1997). Morphological changes of in vitro produced bovine blastocysts after vitrification, in-straw direct rehydration and culture. Mol. Reprod. Dev. 48, 9–17.3.0.CO;2-N>CrossRefGoogle ScholarPubMed
Van Soom, A., Vanroose, G. & de Kruif, A. (2001). Blastocyst evaluation by means of differential staining: a practical approach. Reprod. Dom. Anim. 36, 29–35.CrossRefGoogle ScholarPubMed
Winterberger-Torres, S. & Sevellec, C. (1987). Atlas of the Early Development of the Sheep Embryo. INRA (Institute National de la Reserche Agronomique) Publ. Versailles.Google Scholar