Arias María Elena, Sánchez Raúl, Felmer Ricardo
Laboratory of Reproduction,Centre of Reproductive Biotechnology (CEBIOR-BIOREN),Universidad de La Frontera,Temuco,Chile.
Laboratory of Reproduction,Centre of Reproductive Biotechnology (CEBIOR-BIOREN),Department of Agricultural Sciences and Natural Resources,Faculty of Agriculture and Forestry,Universidad de La Frontera,Montevideo 0870,P.O. Box 54-D,Temuco,Chile.
Zygote. 2016 Oct;24(5):724-32. doi: 10.1017/S0967199416000034. Epub 2016 May 3.
Increasing the efficiency of intracytoplasmic sperm injection (ICSI) in domestic animals has been attempted by many researchers, however embryonic development to the blastocyst stage remains low compared with that of in vitro fertilization (IVF) embryos. One of the main problems observed in cattle is inadequate oocyte activation after ICSI. The present study compared the effect of cycloheximide (CHX), 6-dimethylaminopurine (DMAP), and anisomycin (ANY) on the fertilization rate, development, ploidy and quality of bovine embryos generated by ICSI. Although no differences were observed between treatments in terms of cleavage, higher blastocyst rates were observed for ANY (37.3%) compared with CHX (21.8%, P 0.05) treatments. No differences were observed in the quality of embryos as assessed by the total number of cells, their distribution to the different embryo compartments [inner cell mass (ICM) and trophectoderm (TE)], the proportion of ICM cells to the total cell numbers and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL)-positive cells. Similarly, no differences were observed in the normal ploidy of embryos (56, 67, and 55%) for ANY, CHX and DMAP, respectively. However, higher fertilization rates were observed for ANY (75%) and CHX (87%) treatments compared with DMAP (35%). In conclusion, ANY showed a superior developmental rate compared with CHX treatment. Although no significant differences were observed compared with an improved protocol of DMAP (2Io-DMAP), the lower fertilization rate recorded with DMAP strongly suggests that ANY could be a better alternative for oocyte activation than traditional chemical compounds used currently in ICSI.
许多研究人员尝试提高家畜卵胞浆内单精子注射(ICSI)的效率,然而与体外受精(IVF)胚胎相比,发育至囊胚阶段的胚胎仍然较少。在牛身上观察到的主要问题之一是ICSI后卵母细胞激活不足。本研究比较了环己酰亚胺(CHX)、6-二甲基氨基嘌呤(DMAP)和茴香霉素(ANY)对ICSI产生的牛胚胎的受精率、发育、倍性和质量的影响。虽然在卵裂方面各处理间未观察到差异,但与CHX(21.8%,P<0.05)处理相比,ANY处理的囊胚率更高(37.3%)。通过细胞总数、其在不同胚胎区域[内细胞团(ICM)和滋养外胚层(TE)]的分布、ICM细胞占总细胞数的比例以及末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)阳性细胞评估,胚胎质量未观察到差异。同样,ANY、CHX和DMAP处理的胚胎正常倍性分别为56%、67%和55%,未观察到差异。然而,与DMAP(35%)处理相比,ANY(75%)和CHX(87%)处理的受精率更高。总之,与CHX处理相比,ANY显示出更高的发育率。虽然与改进的DMAP方案(2Io-DMAP)相比未观察到显著差异,但DMAP记录的较低受精率强烈表明,与目前ICSI中使用的传统化合物相比,ANY可能是卵母细胞激活的更好选择。