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Abstract For deeply researching parthenogenetic development of cashmy goat oocytes, and offerring an efficient in vitro activated method for the somatic cell nuclear transfer of cashmy goat, the ovaries from slaughterhouse were used as material and cutted to collect oocyte, then the oocytes were matured in vitro. In this experement the matured oocytes were treated with IA23187＋ 6-dimethylaminopurine(6-DMAP), Ionomycin＋ 6-DMAP, 7% ethanol＋ 6-DMAP, IA23187＋ CHX, Ionomycin＋ CHX and 7% ethanol ＋ cycloheximide (CHX) respectively. Activated oocytes were cultured in vitro in modified synthetic oviductal fluid (mSOF) medium and assessed for cleavage, 8-cell stage (48 h) and the blastocyst stage (7~9 d). The cleavage rates were lower significantly in IA23187＋ 6-DMAP(89.6±4.9%) compared to Ionomycin＋ 6-DMAP(99.1± 0.8%), IA23187＋CHX(97.6±1.2%), Ionomycin＋ CHX (99.3±0.6%) and 7% ethanol＋ CHX(97.5±1.4%)(P＜0.05), but no significantly different with 7% ethanol＋ 6-DMAP(96.3±0.5%, P > 0.05); The 8-cell stage rates were higher significantly in IA23187＋ CHX(81.3±5.8%) and Ionomycin＋CHX (82.5±6.6%) compared to other treatments(48.4± 4.3, 42.5± 1.8, 38.2± 2.1 and 62.5± 2.2%, respectively, P＜0.05); The blastocyst rates were higher significantly in IA23187＋ 6-DMAP(22.8± 2.4%) and Ionomycin＋ 6-DMAP(20.6±4.8%) compared to other treatments (10.5±1.8, 8.4±1.2, 11.8±2.0 and 8.8±2.5%, respectively, P＜0.05); The cell number of blastocysts were not significantly different among treatments 200.0 ±14.0, 169.6 ± 11.4, 187.0 ± 8.8, 201.3±16.5, 179.6±5.2 and 177.6±20.8, respectively, (P > 0.05). All groups of IA23187, Ionomycin and the combination of ethanol and CHX used to activate goat oocyte could obtain higher rate of oocyte cleavage and higher 8-cell rate, the combination of IA23187 and 6-DMAP group and the combination of Ionomycin and 6-DMAP group could obtain higher blastocyst rate, and the blastocyst from different actived group all had higher cell number, the group of either IA23187 or the combination of Ionomycin and 6-DMAP was the best method to activate goat oocyte, and they were also the better in vitro activated method for the technology of somatic cell nuclear transfer, the results of this resarch can offer reference for parthenogenetic development and in vitro activation of goat oocyte.
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Received: 09 November 2010
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