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Survival and ultrastructure of gene-microinjected rabbit embryos after vitrification

  • M. Popelková (a1), P. Chrenek (a2), J. Pivko (a2), A.V. Makarevich (a2), E. Kubovičová (a2) and J. Kačmárik (a1)...

Abstract

Morphological signs of injury and subsequent regeneration following vitrification of either rabbit gene microinjected (Gene-Mi) or intact in vitro cultured embryos derived from in vivo fertilized eggs were evaluated by post-warming recovery in culture and analysed by transmission electron microscopy (TEM). The percentages of vitrified/warmed Gene-Mi embryos that reached the blastocyst stage (69%) and hatched (57%) did not differ significantly from those of intact embryos (78% and 56%, respectively). In contrast, in vitro development of embryos to the blastocyst stage among non-vitrified intact (96%) and Gene-Mi (90%) embryos compared with both the intact vitrified (78%) and Gene-Mi vitrified (69%) groups, as well as hatching rate (94%, 90% vs 56%, 57%, respectively) varied significantly (p<0.001). Observations by TEM showed that the vitrified/warmed intact or Gene-Mi embryos without post-culture displayed severe degenerative changes among their cells. During 24 h of culture a proportion of the embryos were able to regenerate and complete the compaction process. Nevertheless the signs of previous injury were retained, such as swollen cytoplasmic organelles and remaining cellular debris in the perivitelline space. These observations indicate that the procedure of gene Mi does not siginificantly compromise embryo tolerance to cryopreservation and post-warming developmental ability. Severe changes in embryo morphology, observed at the ultrastructural level, can be attributed to a direct influence of the vitrification process rather than to the Mi procedure itself.

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Corresponding author

All correspondence to: M. Popelková, DVM, Clinic of Obstetrics, Gynaecology and Andrology, University of Veterinary Medicine, Komenského 73, 041 81, Košice, Slovak Republic. Tel: +421 55 633 8071. Fax: +421 55 632 3666. e-mail: m.popelkova@lycos.com

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Survival and ultrastructure of gene-microinjected rabbit embryos after vitrification

  • M. Popelková (a1), P. Chrenek (a2), J. Pivko (a2), A.V. Makarevich (a2), E. Kubovičová (a2) and J. Kačmárik (a1)...

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