Parthenogenetic development of in vitro matured bovine oocytes activated by combining methods
1998
Jung, Y.G. (Obihiro Univ. of Agriculture and Veterinary Medicine, Hokkaido (Japan)) | Takeuchi, D. | Fukui, Y.
The present study was undertaken to determine an appropriate method of artificial activation of in vitro matured bovine oocytes on in vitro development rate (DR) to blastocysts. In Experiment 1, in vitro matured (IVM) bovine oocytes were activated once or twice (1 h interval) by 3.5% or 7% ethanol and 2.5 or 5 mu-M Ca-ionophore (A23187), and were treated with cycloheximide and cytochalasin B. The treated oocytes were cultured in synthetic oviduct fluid medium (SOFM) containing 0.8% BSA. The rates of cleavage and development of the treated oocytes were recorded. In the treatment with 3.5% ethanol or 2.5 mu-M Ca-ionophore, the DR of oocytes was significantly (P0.05) higher when a double-activation was given. When the concentration of ethanol and Ca-ionophore increased to 7% and 5 mu-M, however, no significant difference was found between the single- and double-activation. The highest DR (30 or 27%) was achieved when a single treatment with either 7% ethanol or 5 mu-M Ca-ionophore was employed. The cell numbers of the blastocyst derived from treatments of a double-activation with both 3.5 and 7% ethanol were higher (P0.05) than those of each single-activation. There was no significant difference in the cell numbers of blastocyst among the treatments with Ca-ionophore. In Experiment 2, IVM bovine oocytes were activated once with DC-pulse, 5 mu-M Ca-ionophore + DC, or 7% ethanol + DC. The DR of oocytes in the latter two treatments were significantly (P0.05) higher than
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