Czech J. Anim. Sci., 2008, 53(9):388-397 | DOI: 10.17221/336-CJAS

Hypodiploidy as a prominent attributor to chromosomal aneuploidy in transgenic rabbit embryos

S. Roychoudhury1, J. Bulla1, J. Čurlej1,2, P. Chrenek2
1 Department of Animal Physiology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Nitra, Slovak Republic
2 Slovak Agricultural Research Centre, Nitra, Slovak Republic

Transgenic animals play a vital role in basic research, agriculture and pharmaceutical industries. Rabbits have the advantage of other large laboratory species in that they have a short gestation period and yield large numbers of embryos. Production of transgenic rabbits has been directed towards using the rabbit as a model for large domestic animals or as a basic biological model for studying the mammalian gene regulation. In connection with their use, developmental and health disorders have also been reported in genetically modified animals. Random integration of a transgene can disrupt the function or regulation of an endogenous gene, resulting in insertion mutations or chromosomal aneuploidy. Chromosomal abnormalities affect the developmental potential of early embryos and serve as potential predictors of developmental outcome. This study was aimed at analyzing the cytogenetic profile of transgenic rabbit embryos, which is necessary for selecting optimal lines for dissemination in order to eliminate animals with chromosomal aberrations. Conventional Giemsa stained c-metaphase spreads obtained from blastomeres of intact as well as microinjected transgenic (EGFP and hFVIII) and non-transgenic embryos revealed a significantly higher (P < 0.01) rate of aneuploid cells in transgenic rabbits compared to non-transgenic animals. However, microinjection did not seem to influence the rate of aneuploidy, as the incidence of aneuploidy in non-transgenic blastomeres was significantly lower (P < 0.01) in comparison with intact ones (14.3 vs. 73.33%). The findings suggest hypodiploidy as the prominent attributor to the occurrence of aneuploidy. This is the first report of 100% chromosomal aneuploidy in the embryos of both EGFP and hFVIII transgenic rabbits.

Keywords: chromosomal aneuploidy; EGFP; embryo; hFVIII; rabbit; transgenic

Published: September 30, 2008  Show citation

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Roychoudhury S, Bulla J, Čurlej J, Chrenek P. Hypodiploidy as a prominent attributor to chromosomal aneuploidy in transgenic rabbit embryos. Czech J. Anim. Sci. 2008;53(9):388-397. doi: 10.17221/336-CJAS.
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    References

    1. Austin C.R. (1967): Biological mechanisms in aging. Nature, 213, 1018-1019. Go to original source...
    2. Bielanska M.M. (2002): Chromosomal mosaicism in the human preimplantation embryos in vitro. [PhD. Thesis]. McGill University, Canada.
    3. Bronson S.K., Smithies O., Mascarello J.T. (1995): High incidence of XXY and XYY males among the offspring of female chimeras from embryonic stem cells. In: Proceedings of the National Academy of Sciences of the United States of America, USA, 92, 3120-3123. Go to original source...
    4. 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 Research, 14, 417-428. Go to original source... Go to PubMed...
    5. Curlej J., Roychoudhury S., Bulla J., Chrenek P. (2006): Chromosomal analysis of transgenic rabbits. Acta fytotechnica et zootechnica. Special Issue. Year 9, Slovak Agricultural University in Nitra, Slovak Republic, 57-59.
    6. Curlej J., Roychoudhury S., Omelka R., Chrenek P. (2007): Level of chromosomal aneuploidy detected from peripheral blood and bone marrow in transgenic rabbits. Slovak Journal of Animal Science, 40, 5-8.
    7. Diskin M.G. (1986): Embryonic mortality in farm animals: a seminar in the CEC programme of coordination of research. Published by Springer. 292 pp.
    8. Earley E.M. (1976): Growth characteristics, cytogenetic stability and virus susceptibility of cell lines from New Zealand rabbits. Developments in Biological Standardization, 37, 35-39. Go to PubMed...
    9. Fulka H., John J.C.S., Fulka J., Hozak P. (2008): Chromatin in early mammalian embryos: achieving the pluripotent state. Differentiation, 76, 3-14. Go to original source... Go to PubMed...
    10. Gómez M.C., Pope C.E., Lopez M., Dumas C., Giraldo A., Dresser B.L. (2006): Chromosomal aneuploidy in African wildcat somatic cells and cloned embryos. Cloning and Stem Cells, 8, 69-78. Go to original source... Go to PubMed...
    11. Goepfert T.M., McCarthy M., Kittrell F.S., Stephens C., Ullrich R.L., Brinkley B.R., Medina D. (2000): Progesterone facilitates chromosome instability (aneuploidy) in p53 null normal mammary epithelial cells. FASEB Journal, 14, 2221-2229. Go to original source... Go to PubMed...
    12. Hare et al. (1980): Chromosomal abnormalities: A potential quality issue for cloned cattle embryos. In: Slimane-Bureau W.C., King WA.. Cloning and Stem Cells, 4319-4329.
    13. Hughes D.T. (1968): Cytogenetical polymorphism and evolution in mammalian somatic cell populations in vivo and in vitro. Nature, 217, 518-523. Go to original source... Go to PubMed...
    14. Ikawa M., Kominami K., Yoshimura Y., Tanaka K., Nishimune Y., Okabe M. (1995): A rapad and non-invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP). FEBS Letters, 375, 125-128. Go to original source... Go to PubMed...
    15. Karp L.E., Smith W.D. (1975): Experimental production of aneuploidy in mouse oocytes. Gynecology Investigation, 6, 337-341. Go to original source... Go to PubMed...
    16. King W.A. et al. (1979): Embryonic mortality in farm animals: a seminar in the CEC programme of coordination of research (eds.): Sreenan J.M., Diskin M.G.. Published by Springer. 292 pp.
    17. King W.A. et al. (1981): Chromosomal abnormalities: a potential quality issue for cloned cattle embryos. In: Slimane-Bureau W.C. and King W.A.. Cloning and Stem Cells, 4, 319-329. Go to original source... Go to PubMed...
    18. King W.A., Coppola G., Alexander B., Mastromonaco G., Perrault S., Nino-Soto M.I., Pinton A., Joudrey E.M., Betts D.H. (2006): The impact of chromosome alteration on embryo development. Theriogenology, 65, 166-177. Go to original source... Go to PubMed...
    19. Korstanje R., O'Brien P.C.M., Yang F., Rens W., Bosma A.A., van Lith H.A., van Zutphen L.F.M., FergusonSmith M.A. (1999): Complete homology maps of the rabbit (Oryctolagus cuniculus) and human by reciprocal chromosome painting. Animal Cytogenetics and Comparative Mapping, 86, 317-322. Go to original source... Go to PubMed...
    20. Kupriyanov S., Zeh K., Baribault H. (1998): Double pronuclei injection of DNA into zygotes increase yields of transgenic mouse lines. Transgenic Research, 7, 223-226. Go to original source... Go to PubMed...
    21. Ma Sai (1995): Superovulation and chromosomal aberrations. [PhD Thesis]. University of British Columbia, Canada.
    22. Mahmoud G.H.M. (2001): Cytogenetic studies on in vitro fertilization in buffaloes. [PhD thesis]. Faculty of Veterinary Medicine, Cairo University, Egypt.
    23. Mahmoud G.H.M., Hamam A.M., Nawito M.F., Seida A.A.M., Nawar S.M.A. (2002): Cytogenetic analysis of early stage buffalo embryos matured and fertilized in vitro. Egypt Journal of Veterinary Science, 36, 87-98.
    24. Mench J.A. (1999): Ethics, animal welfare, and transgenic farm animals. In: Murray J.D., Anderson G.B., Oberbauer A.M., McGloughlin M.M. (eds.): Transgenic Animals in Agriculture. CABI International, Wallingford, UK, 251-268.
    25. Palmiter R.D., Brinster R.L. (1986): Germ-line transformation of mice. Annual Review of Genetics, 20, 465-499. Go to original source... Go to PubMed...
    26. Parkanyi V. (1981): Utilization of some methods of cytogenetic analysis in the detection of chromosome anomalies in the species Oryctolagus cuniculus. Agriculture, 27, 148-154.
    27. Parkanyi V., Chrenek P., Rafay J., Suvegova K., Jurcik R., Makarevich A.V., Pivko J., Hetenyi L., Paleyanda R.K. (2004): Aneuploidy in the transgenic rabbit. Folia Biologica, 50, 194-199. Go to original source... Go to PubMed...
    28. Roychoudhury S., Bulla J., Curlej J., Chrenek P. (2007): Chromosomal ploidy in transgenic rabbits. Acta Biochimica Polonica, 1, 53-54.
    29. Schumacher A., Fischer B., Agorastos T., Beier H.M. (1993): High frequency of DNA ploidy abnormalities in preimplantation embryos of the rabbit. Molecular Reproduction and Development, 35, 127-133. Go to original source... Go to PubMed...
    30. Shephard S.E., Werner K.L., Schlatter C. (1994): The lacI transgenic mouse mutagenicity assay: quantitative evaluation in comparison to tests for carcinogenicity and cytogenetic damage in vivo. Mutation Research, 306, 119-128. Go to original source... Go to PubMed...
    31. Shi W., Dirim F., Wolf E., Zakhartchenko V., Haaf T. (2004): Methylation reprogramming and chromosomal aneuploidy in in vivo fertilized and cloned rabbit preimplantation embryos. Biology of Reproduction, 71, 340-347. Go to original source... Go to PubMed...
    32. Singh E.L., Hare W.C. (1980): Embryonic mortality in farm animals: a seminar in the CEC programme of coordination of research (eds.): Sreenan J.M., Diskin M.G. Springer. 292 pp.
    33. Slimane W., Heyman Y., Lavergne Y., Humblot P., Renard J.P. (2000): Assessing chromosomal abnormalities in two-cell bovine in vitro-fertilized embryos by using fluorescent in situ hybridization with three different cloned probes. Biology of Reproduction, 62, 628-635. Go to original source... Go to PubMed...
    34. Stromqvist M., Houdebine L.M., Anderson J.O., Edlund A., Johansson T., Viglitta C., Puissant C., Hansson L. (1997): Recombinant human extracellular superoxide dismutase produced in milk of transgenic rabbits. Transgenic Research, 6, 271-278. Go to original source... Go to PubMed...
    35. Susiarjo M., Hassold T.J., Freeman E., Hunt P.A. (2007): Bisphenol A exposure in utero disrupts early oogenesis in the mouse. PLoS Genetic. Available at: doi:10.1371/journal.pgen.0030005 Go to original source... Go to PubMed...
    36. Viuff D., Rickords L., Offenberg H., Hyttel P., Avery B., Greve T., Olsaker I., Williams J.L., Callesen H., Thomson P.D. (1999): A high proportion of bovine blastocysts produced in vitro are mixoploid. Biology of Reproduction, 60, 1273-1278. Go to original source... Go to PubMed...
    37. Ward B.E., Gersen S.L., Carelli M.P., McGuire N.M., Dackowski W.R., Weinstein M., et al. (1993): Rapid prenatal diagnosis of chromosomal aneuploidies by fluorescence in situ hybridization: clinical experience with 4500 specimens. American Journal of Human Genetics, 52, 854-865.
    38. Yang D.P., Rosanoff E.I. (1977): Specific chromosome changes associated with rabbit cell lines cultured in vitro. Cytogenetics and Cell Genetics, 18, 212-230. Go to original source... Go to PubMed...
    39. Zartman D.L., Fechheimer N.S. (1967): Somatic aneuploidy and polyploidy in inbred and linecross cattle. Journal of Animal Science, 26, 252-258. Go to original source... Go to PubMed...

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