Winger Q A, De La Fuente R, King W A, Armstrong D T, Watson A J
Department of Obstetrics and Gynaecology, University of Western Ontario, London, Canada.
Dev Genet. 1997;21(2):160-6. doi: 10.1002/(SICI)1520-6408(1997)21:2<160::AID-DVG5>3.0.CO;2-5.
The present study was conducted to examine the karyotypes of parthenogenetic bovine embryos arising from the application of standard oocyte activation and diploidization methods. Bovine cumulusoocyte complexes were collected and matured in vitro for 24 hr prior to oocyte activation with either 5 microM ionomycin or 7% ethanol for 5 min. Groups of activated oocytes were further treated with 5 micrograms/ml cytochalasin D or 1.9 mM 6-dimethylaminopurine (DMAP) for 6 hr. Cleavage varied significantly (P < .05) among the treatment groups with 68.0% of the ethanol- and DMAP-treated oocytes dividing. Blastocyst development did not vary with 18.4 +/- 2.5% of all treated oocytes progressing to this stage. Blastocyst development did not occur in groups subjected to oocyte activation alone. Blastocysts displayed haploid (2.3%), diploid (11.4%), tetraploid (40.9%), octaploid (4.5%), and mixoploid chromosomal complements (40.9%). Two-cell stage parthenogenotes resulting from ethanol or ionomycin treatment alone displayed haploid (66.7%), diploid (16.7%), tetraploid (4.2%), and mixoploid (12.5%) complements. Our results demonstrate that diploid bovine parthenogenotes arising from these procedures are a minority, with the majority of parthenogenotes displaying polyploid and mixoploid chromosomal complements. The events contributing to these abnormal chromosomal complements occur as early as completion of the first cell cycle, possibly linking these events with the absence of a paternally supplied centrosome.
本研究旨在检测应用标准卵母细胞激活和二倍体化方法产生的孤雌生殖牛胚胎的核型。收集牛卵丘-卵母细胞复合体,在体外成熟24小时,然后用5微摩尔离子霉素或7%乙醇激活卵母细胞5分钟。激活的卵母细胞组再用5微克/毫升细胞松弛素D或1.9毫摩尔6-二甲基氨基嘌呤(DMAP)处理6小时。各处理组的卵裂率差异显著(P <.05),其中经乙醇和DMAP处理的卵母细胞有68.0%发生分裂。囊胚发育情况无差异,所有处理的卵母细胞中有18.4±2.5%发育到该阶段。仅进行卵母细胞激活的组未出现囊胚发育。囊胚显示出单倍体(2.3%)、二倍体(11.4%)、四倍体(40.9%)、八倍体(4.5%)和混合倍体染色体组成(40.9%)。仅经乙醇或离子霉素处理产生的二细胞期孤雌生殖体显示出单倍体(66.7%)、二倍体(16.7%)、四倍体(4.2%)和混合倍体(12.5%)组成。我们的结果表明,通过这些程序产生的二倍体牛孤雌生殖体是少数,大多数孤雌生殖体显示多倍体和混合倍体染色体组成。导致这些异常染色体组成的事件早在第一个细胞周期完成时就已发生,这可能将这些事件与缺乏父源中心体联系起来。