Felmer R, Arias M E
Laboratory of Reproduction, Centre of Reproductive Biotechnology (CEBIOR-BIOREN), Universidad de La Frontera, Temuco, Chile.
Faculty of Agriculture and Forestry, Department of Agricultural Sciences and Natural Resources, Universidad de La Frontera, Temuco, Chile.
Mol Reprod Dev. 2015 Jun;82(6):441-9. doi: 10.1002/mrd.22492. Epub 2015 Apr 29.
We evaluated the potential effect of anisomycin, an antibiotic produced by Streptomyces griseolus, on the parthenogenetic activation of bovine oocytes and reconstructed somatic-cell nuclear transfer (SCNT) embryos. A higher cleavage and blastocyst rate were achieved with anisomycin (70.3% and 27.8%) and 6-dimethylaminopurine (DMAP) (73.3% and 30.2%), relative to oocytes parthenogenetically activated with cycloheximide (CHX) (54.1% and 20.2%). In reconstructed SCNT embryos, a greater proportion of embryos reached the blastocyst stage after anisomycin (32.2%) compared to DMAP (22.3%) and CHX (23.5%) treatment. Furthermore, the quality of embryos-assessed by the total number of cells and the inner cell mass-to-total-cell ratio-was higher with anisomycin (166.2 ± 6.9 and 26.9 ± 1.9) compared to DMAP (135.0 ± 8.7 and 39.4 ± 3.5) and CHX (149.1 ± 8.4 and 36.3 ± 2.5), while a lower percentage of chromosomal abnormalities was observed with anisomycin compared to DMAP and CHX treatments, both in parthenotes (though not significant) and in SCNT embryos (P < 0.05). Therefore, anisomycin can enhance the in vitro developmental potential in parthenotes and reconstructed SCNT embryos, specifically improving the quality of SCNT embryos and decreasing the abnormal ploidy of parthenotes and SCNT embryos compared to the traditional protocols of chemical activation with DMAP and CHX. These results may have important implications for the success of reproductive technologies, including SCNT and intracytoplasmic sperm injection, in different mammalian species.
我们评估了由灰链霉菌产生的抗生素茴香霉素对牛卵母细胞孤雌激活及重构体细胞核移植(SCNT)胚胎的潜在影响。与用放线菌酮(CHX)孤雌激活的卵母细胞(54.1%和20.2%)相比,茴香霉素(70.3%和27.8%)和6-二甲基氨基嘌呤(DMAP)(73.3%和30.2%)处理后的卵裂率和囊胚率更高。在重构的SCNT胚胎中,与DMAP(22.3%)和CHX(23.5%)处理相比,茴香霉素处理后达到囊胚阶段的胚胎比例更高(32.2%)。此外,与DMAP(135.0±8.7和39.4±3.5)和CHX(149.1±8.4和36.3±2.5)相比,茴香霉素处理的胚胎质量(根据细胞总数和内细胞团与总细胞比例评估)更高(166.2±6.9和26.9±1.9),同时在孤雌胚(虽无显著性差异)和SCNT胚胎中,与DMAP和CHX处理相比,茴香霉素处理观察到的染色体异常百分比更低(P<0.05)。因此,与用DMAP和CHX进行化学激活的传统方案相比,茴香霉素可提高孤雌胚和重构SCNT胚胎的体外发育潜力,特别是提高SCNT胚胎的质量,并降低孤雌胚和SCNT胚胎的异常倍性。这些结果可能对包括SCNT和胞浆内单精子注射在内的不同哺乳动物物种生殖技术的成功具有重要意义。