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Quality of rabbit vitrified/thawed transgenic embryos

Published online by Cambridge University Press:  15 August 2011

P. Chrenek ,
Z. Turanová ,
J. Slamečka Jr  and
A. V. Makarevich
P. Chrenek*
Affiliation:
Animal Production Research Centre Nitra, 95141 Luzianky near Nitra, Slovak Republic. Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Slovak Republic.
Z. Turanová
Affiliation:
Animal Production Research Centre Nitra, Hlohovecka 2, 951 41 Luzianky, Slovak Republic.
J. Slamečka Jr
Affiliation:
Animal Production Research Centre Nitra, Hlohovecka 2, 951 41 Luzianky, Slovak Republic. Faculty of Biotechnology and Food Science, Slovak University of Agriculture, Slovak Republic.
A. V. Makarevich
Affiliation:
Animal Production Research Centre Nitra, Hlohovecka 2, 951 41 Luzianky, Slovak Republic.
*
All correspondence to: P. Chrenek. Animal Production Research Centre Nitra, 95141 Luzianky near Nitra, Slovak Republic. Tel: +421 37 6546 289. Fax: +421 37 6546 285. e-mail: [email protected]
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Summary

The aim of our study was to investigate the influence of vitrification on developmental rate and quality (total number of cells, number of blastomeres in inner cell mass (ICM) area, apoptotic index and embryo diameter) of transgenic (carrying an endogenous–hFVIII or exogenous–enhanced green fluorescent protein (EGFP) gene) rabbit embryos. EGFP-positive rabbit embryos were produced under in vitro conditions by the microinjection of foreign genes into the pronucleus of fertilized eggs. The transgenic rabbit embryos with the hFVIII gene were produced by mating homozygous transgenic rabbits and flushing at the single-cell stage. Developmental rate of vitrified/thawed transgenic embryos that reached hatching blastocyst stage (68.00% and 69.00%) and differed significantly (p < 0.001) from those in control embryos (100.00%). Significant difference (p < 0.05) was found in total cell counts between control (117.00 ± 36.00) and vitrified (141.00 ± 34.80) hFVIII-positive embryos. The higher proportion of ICM cells (32.00%) and greatest embryo diameter (130.85 ± 10.90) were found in the control group compared with the transgenic. Ratio of apoptotic cells was significantly higher (p < 0.01) in the control group (2.50%) and vitrified EGFP-positive embryos (2.90%) compared with the vitrified, hFVIII-positive group of embryos (0.70%). Our results demonstrate that neither gene microinjection itself, nor exogenous (EGFP) and endogenous (hFVIII) gene expression interferes with developmental rate and quality of rabbit embryos. However, a combination of microinjection and vitrification significantly decreases (p < 0.001) the survival rate of rabbit embryos.

Keywords

EmbryoQualityRabbitTransgenicVitrification
Type
Research Article
Information
Zygote , Volume 21 , Issue 1 , February 2013 , pp. 53 - 58
Copyright
Copyright © Cambridge University Press 2011

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