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Alteration in ultrastructural morphology of bovine embryos following subzonal microinjection of bovine viral diarrhea virus (BVDV)

Published online by Cambridge University Press:  01 August 2008

E. Kubovičová ,
A. V. Makarevich ,
J. Pivko ,
P. Chrenek ,
P. Grafenau ,
L'. Ríha ,
A. V. Sirotkin  and
F. Louda
E. Kubovičová*
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
A. V. Makarevich
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
J. Pivko
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
P. Chrenek
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
P. Grafenau
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
L'. Ríha
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
A. V. Sirotkin
Affiliation:
Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic.
F. Louda
Affiliation:
Research Institute for Cattle Breeding Ltd., Rapotín, Vikýřovice 78813, Czech Republic.
*
All correspondence to: E. Kubovičová. Slovak Agricultural Research Centre, Hlohovská 2, 949 92 Nitra, Slovak Republic. Tel: +421 37 6546 276. Fax: +421 37 6546 480. e-mail: [email protected]
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Summary

The aim of the present study was to evaluate the development and ultrastructure of preimplantation bovine embryos that were exposed to bovine viral diarrhea virus (BVDV) in vitro.

The embryos were recovered from superovulated and fertilized Holstein-Friesian donor cows on day 6 of the estrous cycle. Compact morulae were microinjected with 20 pl of BVDV suspension (105.16 TCID50/ml viral stock diluted 1:4) under the zona pellucida (ZP), then washed in SOF medium and cultured for 24–48 h. Embryos were evaluated for developmental stages and then processed immunocytochemically for the presence of viral particles, using fluorescent anti-BVDV–FITC conjugate. Ultrastructure of cellular organelles was analysed by transmission electron microscopy (TEM).

After microinjection of BVDV under the ZP, significantly more (p < 0.001) embryos (83.33%) were arrested at the morula stage compared with the intact control (30.33%). Immunocytochemical analysis localized the BVDV–FITC signal inside the microinjected embryos. TEM revealed: (i) the presence of virus-like particles in the dilated endoplasmic reticulum and in cytoplasmic vacuoles of the trophoblast and embryoblast cells; (ii) the loss of microarchitecture: and (iii) abnormal disintegrated nuclei, which lacked reticular structure and the heterochromatin area. In all, the embryo nuclear structure was altered and the microarchitecture of the nucleolus had disappeared when compared with the nuclei from control embryos. Dilatation of the intercellular space and the loss of the intercellular gap junctions were often observed in bovine BVDV-exposed embryos.

These findings provide evidence for the adverse effect of BVDV virus on the development of bovine embryos, which is related to irreversible changes in the ultrastructure of cell organelles.

Keywords

Bovine embryoBovine viral diarrhea virusCell organellesMicroinjectionUltrastructure
Type
Research Article
Information
Zygote , Volume 16 , Issue 3 , August 2008 , pp. 187 - 193
Copyright
Copyright © Cambridge University Press 2008

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