Rooke J A, McEvoy T G, Ashworth C J, Robinson J J, Wilmut I, Young L E, Sinclair K D
Scottish Agricultural College, Craibstone Estate, Bucksburn, Aberdeen AB21 9YA, UK.
Theriogenology. 2007 Feb;67(3):639-47. doi: 10.1016/j.theriogenology.2006.09.040. Epub 2006 Oct 30.
The effects on subsequent fetal development of the presence or absence of serum at different times during IVC of ovine zygotes were studied. Zygotes, recovered from superovulated ewes 36h after intrauterine AI using semen from a single sire, were cultured for 5 days in synthetic oviductal fluid (SOF) media supplemented with either BSA and amino acids (SOF-) or with 10% (v/v) steer serum (SOF+). Serum was present or absent during the first two and last 2 days of IVC giving four treatments (SOF-/SOF-; SOF-/SOF+;SOF+/SOF- and SOF+/SOF+). In total, 224 embryos, including 26 in vivo controls, were transferred singly at day 6 post-AI to synchronous recipients and the products of conception recovered at day 125 of gestation. Presence of serum during IVC had a biphasic effect on embryo development. The inclusion of serum during the first 2 days of IVC retarded early embryo development while the inclusion of serum during the last 2 days of IVC produced more blastocysts by day 6. These effects were independent of each other. The presence of serum during the first 2 days of IVC resulted in increased weights of gravid uterus, placenta, fetus, fetal heart and liver. The incidence of fetuses whose total or organ weights were greater than three standard deviations above the corresponding mean weights of control fetuses was also greater when serum was present during the first 2 days of IVC. However, even when serum was absent throughout IVC there was still an infrequent incidence of fetal weights greater than three standard deviations above the mean for control fetuses. These observations provide evidence that it is the early pre-compaction stages of embryo development that are particularly sensitive to perturbations leading to abnormal fetal development.
研究了在体外培养绵羊受精卵(IVC)的不同时间添加或不添加血清对后续胎儿发育的影响。使用来自单一公羊的精液进行子宫内人工授精(AI)36小时后,从超排母羊中回收的受精卵,在添加了牛血清白蛋白(BSA)和氨基酸的合成输卵管液(SOF)培养基(SOF-)或添加10%(v/v)的犊牛血清的培养基(SOF+)中培养5天。在IVC的前两天和最后两天添加或不添加血清,形成四种处理方式(SOF-/SOF-;SOF-/SOF+;SOF+/SOF-和SOF+/SOF+)。总共224个胚胎,包括26个体内对照胚胎,在AI后第6天单独移植到同期受体中,并在妊娠第125天回收受孕产物。IVC期间血清的存在对胚胎发育有双相影响。IVC前2天添加血清会延迟早期胚胎发育,而IVC最后2天添加血清在第6天时会产生更多囊胚。这些影响相互独立。IVC前2天添加血清会导致妊娠子宫、胎盘、胎儿、胎儿心脏和肝脏重量增加。当IVC前2天添加血清时,总重量或器官重量大于对照胎儿相应平均重量三个标准差以上的胎儿发生率也更高。然而,即使在整个IVC过程中都不添加血清,胎儿重量大于对照胎儿平均值三个标准差以上的情况仍偶尔发生。这些观察结果提供了证据,表明胚胎发育的早期致密化前阶段对导致胎儿发育异常的干扰特别敏感。