Hostname: page-component-848d4c4894-xm8r8 Total loading time: 0 Render date: 2024-06-22T10:31:10.007Z Has data issue: false hasContentIssue false
  • English
  • Français

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: chrenekp@yahoo.com
Get access Rights & Permissions [Opens in a new window]

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

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

Brison, D.R. & Schultz, R.M. (1997). Apoptosis during mouse blastocyst formation: evidence for a role for survival factors including TGF-α. Biol. Reprod. 56, 1088–96.CrossRefGoogle Scholar
Chrenek, P. & Makarevich, A.V. (2005). Production of rabbit chimeric embryos by aggregation of zona-free nuclear transfer blastomeres. Zygote 13, 39–44.CrossRefGoogle ScholarPubMed
Chrenek, P., Vašíček, D., Makarevich, A., Uhrin, P., Petrovičová, I., Lubon, H., Binder, B.R. & Bulla, J. (2002). Integration and expression of the WAP-hPC gene in three generations of transgenic rabbits. Czech J. Anim. Sci. 47, 45–9.Google Scholar
Chrenek, P., Vasicek, D., Makarevich, A.V., Jurcik, R., Suvegova, K., Parkanyi, V., Bauer, M., Rafay, J., Batorova, A. & Paleyanda, R.K. (2005). Increased transgene integration efficiency upon microinjection of DNA into both pronuclei of rabbit embryos. Transgenic Res. 14, 417–28.CrossRefGoogle ScholarPubMed
Chrenek, P., Makarevich, A.V., Bauer, M. & Jurcik, R. (2008). Development rate and allocation of transgenic cells in rabbit chimeric embryos. Zygote 16, 87–91.CrossRefGoogle ScholarPubMed
Dobrinski, J.R. (2002). Advancements in cryopreservation of domestic animal embryos. Theriogenology 57, 285–302.CrossRefGoogle 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
Fabian, D., Makarevich, A.V., Chrenek, P., Bukovská, A. & Koppel, J. (2007). Chronological appearance of spontaneous and induced apoptosis during preimplantation development of rabbit and mouse embryos. Theriogenology 68, 1271–81.CrossRefGoogle ScholarPubMed
Fouladi-Nashta, A.A., Alberio, R., Kafi, M., Nicholas, B., Campbell, K.H.S. & Webb, R. (2005). Differential staining combined with TUNEL labeling to detect apoptosis in preimplantation bovine embryos. Reprod. Biomed. Online 4, 497–502.CrossRefGoogle Scholar
Gutierrez-Adan, A., Rizos, D., Fair, T., Moneira, P.N., Pintado, B., de la Fuente, J., Boland, M.P. & Lonergan, P. (2004). Effect of speed of development on mRNA expression pattern in early bovine embryos cultured in vivo or in vitro. Mol. Reprod. Dev. 68, 441–448.CrossRefGoogle ScholarPubMed
Hardy, K. (1999). Apoptosis in the human embryo. Rev. Reprod. 4, 125–34.CrossRefGoogle ScholarPubMed
Hsiao-Sheng, L., Ming-Shion, J., Chao-Kai, C.H., Ping-Hong, C.H. & Nir-Jihn, K. (1999). Is green fluorescent protein toxic to living cells? Biochem. Biophys. Res. Comm. 260, 712–7.Google Scholar
Jurisicova, A. & Acton, B.M. (2004). Deadly decisions the role of genes regulating programmed cell death in human preimplantation embryo development. Reproduction 128, 281–91.CrossRefGoogle ScholarPubMed
Kasai, M., Hamaguchi, Y., Zhu, S.E., Miyake, T., Sakurai, T. & Machida, T. (1992). High survival of rabbit morulae after vitrification in an ethylene glycol-based solution by a simple method. Biol. Reprod. 46, 1042–6.CrossRefGoogle Scholar
Makarevich, A.V., Chrenek, P., Zilka, N., Pivko, J. & Bulla, J. (2005). Preimplantation development and viability of in vitro cultured rabbit embryos derived from in vivo fertilized gene–microinjected eggs: apoptosis and ultrastructure analyses. Zygote 13, 125–37.CrossRefGoogle ScholarPubMed
Makarevich, A.V., Chrenek, P. & Fľak, P. (2006). The influence of microinjection of foreign gene into the pronucleus of fertilized egg on the preimplantation development, cell number and diameter of rabbit embryos. Asian-Aust. J. Anim. Sci. 19, 1–5.Google Scholar
Makarevich, A.V., Chrenek, P., Olexíková, L., Popelková, M., Turanová, Z., Ostró, A. & Pivko, J. (2008). Post-thaw survival, cell death and actin cytoskeleton in gene-microinjected rabbit embryos after vitrification. Theriogenology 70, 675–81.CrossRefGoogle ScholarPubMed
Özkavukcu, S. & Erdemli, E. 2002. Cryopreservation: basic knowledge and biophysical effects. J. Ankara Med. Sch. 4, 187–96.Google Scholar
Papis, K., Sypecka, J., Korwin-Kossakowski, M., Wenta-Muchalska, E. & Bilska, B. (2005). Banking of embryos of mutated, paralytic tremor rabbit by means of vitrification. Lab. Anim. 39, 284–9.CrossRefGoogle ScholarPubMed
Popelková, M., Chrenek, P., Pivko, J., Makarevich, A.V., Kubovičová, E. & Kačmárik, J. (2005). Survival and ultrastructure of gene – microinjected rabbit embryos after vitrification. Zygote 13, 283–93.CrossRefGoogle ScholarPubMed
Popelková, M., Turanová, Z., Koprdová, L., Ostró, A., Toporcerová, S., Makarevich, A.V. & Chrenek, P. (2008). Effect of vitrification technique and assisted hatching on rabbit embryo developmental rate. Zygote 17, 57–61.CrossRefGoogle ScholarPubMed
Roychoudhury, S., Bulla, J., Curlej, J. & Chrenek, P. (2008). Hypodiploidy as a prominent attributor to chromosomal aneuploidy in transgenic rabbit embryos. Czech J. Anim. Sci. 53, 388–97.CrossRefGoogle Scholar
Schwartzman, R.A. & Cidlowski, J.A. (1993). Apoptosis: the biochemistry and molecular biology of programmed cell death. Endocrine Rev. 14, 133–51.Google ScholarPubMed
Voss, A.K., Sandmoller, A., Suske, G., Strojek, R.M., Beato, M. & Hahn, J. (1990). A comparison of mouse and rabbit embryos for the production of transgenic animals by pronuclear microinjection. Theriogenology 34, 813–24.CrossRefGoogle ScholarPubMed
Warner, C.M., Exley, G.E., McElhinny, A.S. & Tang, C. (1998). Genetic regulation of preimplantation mouse embryo survival. J. Exp. Zool. 282, 272–9.3.0.CO;2-U>CrossRefGoogle ScholarPubMed