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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 ,
G. Marsicano ,
J.-E. Fléchon  and
C. Galli
D. Shehu
Affiliation:
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
G. Marsicano
Affiliation:
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
J.-E. Fléchon*
Affiliation:
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
C. Galli
Affiliation:
CIZ Srl, Cremona, Italy, and INRA, Jouy en Josas, France.
*
J.-E. Fléchon, Biologie Cellulaire et Moléculaire, INRA 78352 Jouy en Josas cedex, France.
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Summary

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.

Keywords

Cell junctionsCytoskeletonExtracellular matrixImmunofluorescencePreimplantation bovine embryo
Type
Article
Information
Zygote , Volume 4 , Issue 2 , May 1996 , pp. 109 - 121
Copyright
Copyright © Cambridge University Press 1996

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