Artificial activation of bovine and equine oocytes with cycloheximide, roscovitine, strontium, or 6-dimethylaminopurine in low or high calcium concentrations
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Data
2014-08-01
Autores
Fernandes, Claudia Barbosa
Devito, Liani Gasparini [UNESP]
Martins, Lilian Rigatto [UNESP]
Pria Blanco, Ieda Dala [UNESP]
Lima Neto, Joao Ferreria de [UNESP]
Tsuribe, Patricia Myakawa
Pereria Goncalves, Camila Gabriela
Landim-Alvarenga, Fernanda da Cruz [UNESP]
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Editor
Cambridge University Press
Resumo
Knowledge on parthenogenetic activation of oocytes is important to improve the efficiency of nuclear transfer (NT) and intracytoplasmic sperm injection (ICSI) because artificial activation of oocyte (AOA) is an essential step to achieve embryo production. Although different procedures for AOA have been established, the efficiency of in vitro production of embryos remains low, especially in equines and Bos taurus bovines. In an attempt to improve the techniques of NT and ICSI in bovine and equine species, we tested different combinations of drugs that had different mechanisms of action for the parthenogenetic activation of oocytes in these animals. The oocytes were collected, in vitro matured for 24 to 30 h and activated artificially, in the presence of low or high concentrations of calcium, with combinations of calcium ionophore (ionomycin) with cycloheximide, roscovitine, strontium, or 6-dimethylaminopurine (6-DMAP). For assessment of activation rates, oocytes were stained with Hoechst 33342 and observed under an inverted microscope. We showed that all combinations of drugs were equally efficient in activating bovine oocytes, with the best results obtained when high concentrations of calcium were adopted. For equine oocytes, high concentrations of calcium were not beneficial for the parthenogenetic activation and the combination of ionomycin with either 6-DMAP or roscovitine was effective in inducing artificial activation of oocytes. We believe that our preliminary findings provide some clues for the development of a better AOA protocol to be used with these species.
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Palavras-chave
Cloning, ICSI, Nuclear transfer, Parthenogenetic activation
Como citar
Zygote. New York: Cambridge Univ Press, v. 22, n. 3, p. 387-394, 2014.