Department of Animal Sciences and D.H. Barron Reproductive and Perinatal Biology Research Program, Genetics Institute, University of Florida, Gainesville, FL 32611-0910, USA.
Campo Experimental Centro Altos de Jalisco, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias, Tepatitlán de Morelos, Jalisco, 47600, México.
J Anim Sci. 2021 Aug 1;99(8). doi: 10.1093/jas/skab180.
Alterations in the environment of the preimplantation embryo can affect competence to establish pregnancy and phenotype of resultant calves. In this study, the bovine embryo produced in vitro was used to evaluate postnatal programming actions of the embryokine colony-stimulating factor 2 (CSF2) and serum, which is a common additive of culture media. Oocytes were collected by ovum pick up from Brahman donors and fertilized with semen from Brahman bulls. Embryos were randomly assigned to one of the three treatments: vehicle, CSF2 10 ng/mL, or 1% (v/v) serum. Treatments were added to the culture medium from day 5 to 7 after fertilization. Blastocysts were harvested on day 7 and transferred into crossbred recipients. Postnatal body growth and Longissimus dorsi muscle characteristics of the resultant calves were measured. The percent of cleaved embryos becoming blastocysts was increased by serum and, to a lesser extent, CSF2. Treatment did not affect survival after embryo transfer but gestation length was shortest for pregnancies established with serum-treated embryos. Treatment did not significantly affect postnatal body weight or growth. At 3 mo of age, CSF2 calves had lower fat content in the Longissimus dorsi muscle and less subcutaneous fat over the muscle than vehicle calves. There was a tendency for cross-sectional area of the muscle to be smaller for serum calves than vehicle calves. Results confirm the importance of the preimplantation period as a window to modulate postnatal phenotype of resultant calves. In particular, CSF2 exerted actions during the preimplantation period to program characteristics of accumulation of intramuscular and subcutaneous fat of resultant calves. The use of a low serum concentration in culture medium from day 5 to 7 of development can increase the yield of transferrable embryos without causing serious negative consequences for the offspring.
胚胎植入前环境的改变会影响胚胎着床能力和后代牛的表型。本研究以体外生产的牛胚胎为模型,评估胚胎集落刺激因子 2(CSF2)和血清这两种常见培养液添加剂的后生编程作用。卵母细胞通过从婆罗门牛供体中吸取卵母细胞进行收集,并与婆罗门公牛的精液进行受精。胚胎被随机分配到以下三种处理之一:载体、10ng/ml CSF2 或 1%(v/v)血清。处理从受精后第 5 天至第 7 天添加到培养液中。第 7 天收获囊胚并移植到杂交受体中。测量所得小牛的产后体生长和背最长肌特征。血清和 CSF2 均能增加胚胎的卵裂率,使胚胎发育为囊胚的比例增加,但 CSF2 的作用较小。处理并不影响胚胎移植后的存活率,但用血清处理胚胎建立的妊娠,其妊娠期最短。处理对产后体重或生长没有显著影响。在 3 月龄时,CSF2 小牛的背最长肌脂肪含量较低,肌肉上的皮下脂肪较少,而载体小牛的脂肪含量较高。血清小牛的肌肉横截面积比载体小牛的更小,有这种趋势。结果证实,着床前时期是调节后代牛产后表型的一个重要窗口。特别是,CSF2 在着床前时期发挥作用,调节了后代牛肌肉内和皮下脂肪积累的特征。在发育的第 5 天至第 7 天的培养液中使用低浓度血清,可以增加可转移胚胎的产量,而不会对后代产生严重的负面影响。