title: Effects of superovulation, culture and microinjection on development of rabbit embryos in vitro.
abstract: Factors influencing the developmental potential of cultured rabbit zygotes and their ability to incorporate and integrate the WAP-hPC (human protein C) gene were investigated. Rabbit zygotes (n = 1053) were recovered from both superovulated and nontreated New Zealand White females. The hormonal treatment of rabbit donors resulted in a doubling of the number of recovered ova per donor when compared with the nontreated group (18 vs 9 ova). However, the quality of recovered zygotes (presence of both pronuclei) was significantly better in the nontreated group (99 vs 88%, Experiment 1). The effect of various culture media on the development of rabbit zygotes in vitro was evaluated after incubation under CO2-free conditions (Experiment 2). In serum-free, growth factor-supplemented medium (BSEITS, DME/F12, 1.5% BSA, EGF, insulin, transferrin and sodium selenite) the percentage of morula/blastocyst stage embryos was significantly higher (88%) than in DME/FCS, (DME/F12, 10% fetal calf serum, 59%) or the control group (DME/F12, 1.5% BSA, 25%). In Experiment 3, zygotes were microinjected with the WAP-hPC gene and were examined after 72 h of culture. Zygote cleavage and the percentage of morula/blastocyst stage intact embryos were higher (79 and 58%, respectively) than in microinjected embryos (31.0 and 21.5%, respectively). Summarized data of the PCR assay of microinjected zygotes demonstrated positive signals for the integration of the WAP-hPC gene in 6.6% (34 of 515) of all the microinjected zygotes.
authors.affiliation: Research Institute of Animal Production, Nitra, Slovak Republic. chrenekp@vuzv.sk
authors.forename: P
authors.initials: P
authors.lastname: Chrenek
authors.forename: A
authors.initials: A
authors.lastname: Makarevich
authors.forename: D
authors.initials: D
authors.lastname: Vasícek
authors.forename: J
authors.initials: J
authors.lastname: Laurincík
authors.forename: J
authors.initials: J
authors.lastname: Bulla
authors.forename: T
authors.initials: T
authors.lastname: Gajarská
authors.forename: J
authors.initials: J
authors.lastname: Rafay
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000502:physiology; D003470:Culture Media; D046508:Culture Techniques; D004622:Embryo, Mammalian / Q000502:physiology*; D005314:Embryonic and Fetal Development; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D008845:Microinjections; D009028:Morula / Q000502:physiology; D011247:Pregnancy; D011486:Protein C / Q000235:genetics; D011817:Rabbits / Q000196:embryology*; D013480:Superovulation; D015053:Zygote / Q000502:physiology
chemical_list: D003470:Culture Media; D011486:Protein C