Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos.
Effect of activation treatments on actin filament distribution and in vitro development of miniature pig somatic cell nuclear transfer embryos.
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DOI：
10.1262/jrd.18162
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发表时间：
2007-08
期刊：
影响因子：
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通讯作者：
K. Yamanaka;S. Sugimura;T. Wakai;T. Shoji;J. Kobayashi;H. Sasada;E. Sato
中科院分区：
文献类型：
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作者：
K. Yamanaka;S. Sugimura;T. Wakai;T. Shoji;J. Kobayashi;H. Sasada;E. Sato
本研究探讨了激活处理对小型猪体细胞核移植（SCNT）胚胎肌动蛋白丝分布和体外发育的影响。我们将离子霉素（ION）、电刺激（ES）和放线菌酮处理（CH）三种活化方法相结合，制备了七种活化处理（ION、ES、CH、ION + CH、ION + ES、ES + CH和ION + ES + CH）。首先，我们研究了卵母细胞的激活率和孤雌体的体外发育。ION、ES、CH、ION + CH、ION + ES、ES + CH和ION + ES + CH组的卵母细胞激活率分别为42.9、51.3、0.0、82.1、80.6、78.1和78.6%，联合处理组的激活率显著高于单一处理组（P<0.05）。虽然联合处理组的激活率没有显著差异，但ION + CH处理组的囊胚发育率（36.1%）显著高于其他联合处理组（14.6-24.7%）（P<0.05）。随后，我们研究了SCNT胚胎中微丝的体外发育和分布。ION + CH处理组的SCNT胚胎囊胚发育率（11.3%）显著高于ES和ION + ES + CH处理组（分别为4.5%和5.2%）（P<0.05）。ION + CH激活的SCNT胚胎中正常肌动蛋白丝分布率显著高于ES或ION + ES + CH激活的SCNT胚胎（63.3% vs.46.8%或46.4%）（P<0.05）。此外，ION + CH激活的SCNT胚胎的碎裂率明显低于ION + ES + CH激活的SCNT胚胎（14.9%比26.1%）（P<0.05）。目前的研究结果表明，离子霉素结合放线菌酮的活化处理可以避免对微丝的物理损伤，并导致小型猪SCNT胚胎的后续发育得到改善。
In the present study, we investigated the effect of activation treatments on the actin filament distribution and in vitro development of somatic cell nuclear transfer (SCNT) embryos in miniature pigs. We combined three activation methods, ionomycin (ION), electrical stimulation (ES), and cycloheximide treatment (CH), to prepare seven activation treatments (ION, ES, CH, ION + CH, ION + ES, ES + CH and ION + ES + CH). First, we investigated the activation rate of oocytes and in vitro development of parthenotes. The activation rates of the oocytes in the ION, ES, CH, ION + CH, ION + ES, ES + CH, and ION + ES + CH groups were 42.9, 51.3, 0.0, 82.1, 80.6, 78.1 and 78.6%, respectively, showing that the rates of the combined treatment groups were significantly higher (P<0.05) than those of the single treatment groups. Although there were no significant differences in the activation rates of the combined treatment groups, the developmental rate to blastocysts in the ION + CH treatment group (36.1%) was significantly higher (P<0.05) than the other combined treatment groups (14.6-24.7%). Subsequently, we investigated the in vitro development and distribution of microfilaments in SCNT embryos. The developmental rate to blastcysts of the SCNT embryos in the ION + CH treatment group (11.3%) was significantly higher (P<0.05) than in the ES and ION + ES + CH treatment groups (4.5 and 5.2%, respectively). The rate of normal actin filament distribution in the SCNT embryos activated with ION + CH was significantly higher (P<0.05) than those activated with ES or ION + ES + CH treatment (63.3 vs. 46.8 or 46.4%). In addition, the fragmentation rate of the SCNT embryos activated with ION + CH was significantly lower (P<0.05) than those activated with ION + ES + CH (14.9 vs. 26.1%). The present results suggest that an activation treatment of ionomycin combined with cycloheximide may avoid physical damage to microfilaments and result in improved subsequent development of miniature pig SCNT embryos.