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Developmentally regulated markers of in vitro-produced preimplantation bovine embroys

Published online by Cambridge University Press:  26 September 2008

D. Shehu
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
G. Marsicano
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
J.-E. Fléchon
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
C. Galli
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.


Expression of various developmentally regulated markers was screened throughout the preimplantation stages of in vitro-derived bovine embryos. This was done by investigating the distribution of several nuclear, cytoplasmic and extracellular proteins by means of immunofluorescence microscopy. While lamin B appeared as a constitutive component of nuclei of all preimplantation stages, lamins A/C had a stage-related distribution. The early cleavage stage nuclei contained lamins A/C which generally disappeared in the following stages, with the possible exception of a few positive nuclei in the morula and early blastocyst stage. In the expanded blastocyst stage the nuclei of trophectoderm cells became positive while no positivity was observed in the inner cell mass cells. Starting from day 6, the appearance and/or polarised distribution of various cytoskeletal and cytoskeleton-related components such as Factin, α-catenin and E-cadherin gave an insight into the timing of events related to compaction of bovine e bryos. Compaction was correlated with the first differentiation event, i.e. the formation of trophectoderm; this is the first embryonic epithelium, characterised by cytokeratins and desmoplakin. Extracellular fibronectin was first detected in the early blastocyst stage shortly before the morphological differentiation of primitive endoderm, and in the later stages it was localised at the interface between trophectoderm and extraembryonic endoderm. Laminin and collagen IV were expressed by the endoderm cells and contributed to the extracellular matrix underlying the trophectoderm. This study is a first attempt to characterise the cells of in vitro-derived bovine embryos valid for cell line derivation.

Copyright © Cambridge University Press 1996

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Betteridge, K.J. & Flèchon, J.E. (1988). The anatomy and physiology of pre–attachment bovine embryos. Theriogenology 29, 155–86.CrossRefGoogle Scholar
Chisholm, J.C. & Houliston, E. (1987). Cytokeratin filament assembly in the preimplantation mouse embryo. Development. 101, 565–82.Google ScholarPubMed
Collins, J.E. & Fleming, T.P. (1995). Epithelial differentiation in the mouse preimplantation embryo: making adhesive cell contacts for the first time. Trends Biochem. Sci. 220, 307–12.CrossRefGoogle Scholar
Cowin, P., Kaprell, H.P. & Franke, W.W. (1985). The complement of desmosomal plaque proteins in different cell types. J. Cell Biol. 101, 11442–54.CrossRefGoogle ScholarPubMed
Enders, A.C., Given, R.L. & Schlafke, S., (1978). Differentiation and migration of endoderin in the rat and mouse at implantation. Anat. Rec. 190, 6577.CrossRefGoogle ScholarPubMed
Eyestone, W.H., Liebfried-Rutledge, L., Northey, D.L., Gillman, B.G. & First, N.L. (1987). Culture of one- and two-cell bovine embryos to the blastocyst stage in the ovine oviduct. Theriogenology 28, 17.CrossRefGoogle ScholarPubMed
Fléchon, J.E. (1994). Request for a consensus on the definition of putative embryonic stem cells. Mol. Reprod. Dev. 41, 274.CrossRefGoogle Scholar
Fleming, T.P., Garrod, R. & Elsmore, A.J. (1991). Desmosome biogenesis in the mouse preimplantation embryo. Development 112, 527–39.Google ScholarPubMed
Fleming, T.P., Butler, L., Lei, X., Collins, J., Javed, Q., Sheth, B., Stoddart, N., Wild, A. & Hay, M. (1994). Molecular maturation of cell adhesion systems during mouse early development. Histochemistry 101, 17.CrossRefGoogle ScholarPubMed
Galli, C. & Lazzari, G. (1994). Large scale production of bovine embryos. In: Proceedings of the First European Conference on Progress in Embryo Technology and Genetic Engineering in Cattle and Sheep Breeding. Krakòw, Poland, pp. 111–16.Google Scholar
Galli, C., Duchi, R. & Lazzari, G. (1994a). Production of purebred embryos from beef cattle by in vitro embryo technology. Theriogenology 41, 20.CrossRefGoogle Scholar
Galli, C., Lazzari, G., Fléchon, J.E. & Moor, R.M. (1994b). Embryonic stem cells in farm animals. Zygote 2, 385–9.CrossRefGoogle ScholarPubMed
Guilly, M.-N., Danon, F., Brouet, J.-C., Bomens, M. & Courvalin, J.-C. (1987). Autoantibodies to nuclear lamin B in a patient with thrombopenia. Eur. J. Cell Biol. 43, 266–72.Google Scholar
Hanzel, D., Reggio, H., Bretcher, A., Forte, J.G. & Mangeat, P. (1991). The secretion stimulated 80K phosphoprotein of parietal cells is ezrin, and has the properties of a membrane cytoskeletal linker in the induced apical microvilli. EMBO J. 10, 2363–73.Google ScholarPubMed
Häger, T.H., Grund, C., Franke, W.W. & Krohne, G. (1991). Immunolocalization of lamins in the thick nuclear lamina of human synovial cells. Eur. J. Cell Biol. 54, 150–6.Google Scholar
Holthäfer, H., Miettinen, A., Paasivuo, R., Lehto, V.P., Linster, E., Aefthan, O. & Virtonen, I. (1983). Cellular origin and differentiation of renal carcinomas. Lab, Invest. 49, 317–26.Google Scholar
Johnson, M.H. & Maro, B. (1984). The distribution of cytoplasmic actin in mouse 8–cell blastomeres. I. Embryol. Exp. Morphol. 90, 311–34.Google Scholar
Johnson, M.H. & Maro, B. (1986). Time and space in the mouse early embryo: a cell biological approach to cell diversification. In Experimental Approaches to Mammalian Embryonic Development, ed. J., Rossant & R.A., Pedersen, pp. 3566. Cambridge: Cambridge University Press.Google Scholar
Kidder, G.M. & McLachlin, J.R. (1985). Timing of transcription and protein synthesis in preimplantation mouse embryos. J. Embryol. Exp. Morphol. 89, 223–34.Google Scholar
Kopečný, V. (1989). High–resolution autoradiographic studies of comparative nucleologenesis and genome reactivation during early embryogenesis in pig, man and cattle. Reprod. Nutr. Dev. 29, 589600.CrossRefGoogle ScholarPubMed
Krohne, G. & Benavente, R. (1986). The nuclear lamins: a multigene family of proteins in evolution and differentiation. Exp. Cell Res. 16, 110.CrossRefGoogle Scholar
Lehtonen, E. & Reima, I. (1986). Changes in the distribution of vinculin during preimplantation mouse development. Differentiation 32, 125–34.CrossRefGoogle ScholarPubMed
Lehtonen, E., Ordónez, G. & Reima, I. (1988). Cytoskeleton in the preimplantation mouse development. Cell Differ. 24, 165–78.CrossRefGoogle ScholarPubMed
Massip, A., Mulnard, J., Huygens, R., Hanzen, C., Van, der, Zwalmen, P. & Ectors, F. (1981). Ultrastructure of the cow blastocyst. J. Submicrosc. Cytol. 13, 3140.Google Scholar
McLaren, A. (1982). The embryo. In Reproduction in Mammals, vol. 2, Embryonic and Fetal Development, ed. C.R., Austin & R.V., Short, pp. 126. Cambridge: Cambridge University Press.Google Scholar
McLaren, A. & Smith, R. (1977). Functional test of tight junctions in the mouse blastocyst. Nature 267, 351–3.CrossRefGoogle ScholarPubMed
Mohr, L.R. & Trounson, A.O. (1981). Structural changes associated with freezing of bovine embryos. Biol. Reprod. 26, 787–90.Google Scholar
Nagafuchi, A. & Tsukita, S. (1994). The loss of the expression of α-catenin, the 120 kD cadherin associated protein, in central nervous tissues during development. Dev. Growth Differ. 36, 5971.CrossRefGoogle Scholar
Papaioannou, V.E. & Ebert, K.M. (1988). The preimplantation pig embryo: cell number and allocation of trophectoderm and inner cell mass of the blastocyst in vivo and in vitro. Development. 102, 793803.Google ScholarPubMed
Pollard, J.W. & Leibo, S.P. (1993). Comparative cryobiology of in vitro– and in vivo-derived bovine embryos. Theriogenology. 39, 287.CrossRefGoogle Scholar
Prather, R.S., Sims, M.M., Maul, G.G., First, N.L. & Shatten, G. (1989). Nuclear lamin antigens are developmentally regulated during porcine and bovine embryogenesis. Biol. Reprod. 41, 123–32.CrossRefGoogle ScholarPubMed
Pratt, H.P.M., Ziomek, C.A., Reeve, W.J.D. & Johnson, M.H. (1982). Compaction of the mouse embryo: an analysis of its components. J. Embryol. Exp. Morphol. 70,113–32.Google ScholarPubMed
Reima, I. (1990). Maintenance of compaction and adherent-type junctions in mouse morula-stage embryos. Cell Differ. Dev. 29, 143–53.CrossRefGoogle ScholarPubMed
Reima, I. & Lehtonen, E. (1985). Localization of nonerythroid spectrin and actin in mouse oocytes and preimplantation embryos. Differentiation. 30, 6875.CrossRefGoogle ScholarPubMed
Reima, I., Lehtonen, E., Virtanen, I. & Fléchon, J.E. (1993). The cytoskeleton and associated proteins during cleavage, compaction and blastocyst differentiation in the pig. Differentiation. 54, 3545.CrossRefGoogle ScholarPubMed
Richoux, V., Darribère, T., Boucaut, J.C., Fléchon, J.E. & Thiéry, J.P. (1989). Distribution of fibronectins and laminin in the early pig embryo. Anat. Rec. 223, 7281.CrossRefGoogle ScholarPubMed
Rieger, D., Loskutoff, N.M. & Betteridge, K.J. (1992). Developmentally related changes in the metabolism of glucose and glutamine by cattle embryos produced and cultured in vitro. J. Reprod. Fertil. 95. 585–95.CrossRefGoogle Scholar
Róber, R.A., Weber, K. & Osborn, M. (1989). Differential timing of nuclear lamin A/C expression in the various organs of the mouse embryos and the young animal: a developmental study. Development 105, 365–78.Google Scholar
Stevenson, B.R., Siciliano, J.D., Mooseher, M.F. & Goodenough, D.A. (1986). Identification of ZO-l: a high molecular weight polypeptide associated with the tight junctions (zonula occludens) in a variety of epithelia. J. Cell Biol. 103, 755–66.CrossRefGoogle Scholar
Stroband, H.W.J., Taverne, N. & Bogaard, M. (1984). The pig blastocyst its ultrastructure and the uptake of protein macromolecules. Cell Tissue Res. 253,347–56.Google Scholar
Van Stekelenburg-Hamers, A.E.P., Van Achterberg, T.A.E., Rebel, H.G., Fléchon, J.E., Campbell, K.H.S., Weima, S.M. & Mummery, C.L. (1995). Isolation and characterization of permanent cell lines from inner cell mass cells of bovine blastocysts. Mol. Reprod. Dev. 40, 444–54.CrossRefGoogle ScholarPubMed
Vartio, T., Laitinen, L., Närvänen, O., Cutolo, M., Thomell, L.E., Zardi, L. & Virtanen, I. (1987). Differential expression of the ED sequence-containing form of cellular fibronectin in embryonic and adult human tissues. J. Cell Sci. 88, 419–30.Google ScholarPubMed
Wartiovaara, J., Leivo, I. & Vaheri, A. (1979). Expression of the cell surface associated glycoprotein, fibronectin, in the early mouse embryo. Dev. Biol. 69, 247557.CrossRefGoogle ScholarPubMed
Ylänne, J. & Virtanen, I. (1989). The Mr 140000 fibronectins complex in normal and virus-transformed human fibroblast and in fibrosarcoma cells: identical localization and function. Int. J. Cancer. 43, 1126–36.CrossRefGoogle ScholarPubMed
Ziomek, C.A. & Johnson, M.H. (1980). Cell surface interactions induces polarization of mouse 8-cell blastomeres at compaction. Cell. 21, 935–42.CrossRefGoogle Scholar
Ziomek, C.A. & Johnson, M.H. (1981). Properties of polar and apolar cells from the 16-cell mouse morula. Rouxs Arch. Dev. Biol. 190, 287–96.CrossRefGoogle ScholarPubMed

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