摘要
Using sex-sorted semen to produce offspring of desired sex is associated with reduced developmental competence in vitro and lower fertility rates in vivo. The objectives of the present study were to investigate the effects of exogenous follistatin supplementation on the developmental competence of bovine embryos produced with sex-sorted semen and possible link between TGF-β regulated pathways and embryotrophic actions of follistatin. Effects of follistatin on expression of cell lineage markers (CDX2 and Nanog) and downstream targets of SMAD signaling (CTGF, ID1, ID2 and ID3) and AKT phosphorylation were investigated. Follistatin was supplemented during the initial 72 h of embryo culture. Exogenous follistatin restored the in vitro developmental competence of embryos produced with sex-sorted semen to the levels of control embryos produced with unsorted semen, and comparable results were obtained using sorted semen from three different bulls. The mRNA abundance for SMAD signaling downstream target genes, CTGF (SMAD 2/3 pathway) and ID2 (SMAD 1/5 pathway), was lower in blastocysts produced using sex-sorted versus unsorted semen, but mRNA levels for CDX2, NANOG, ID1 and ID3 were similar in both groups. Follistatin supplementation restored CTGF and ID2 mRNA in blastocysts produced using sex-sorted semen to levels of control embryos. Moreover, levels of phosphorylated (p)AKT (Ser-473 and Thr-308) were similar in embryos derived from sex-sorted and unsorted semen, but follistatin treatment increased pAKT levels in both groups. Taken together, results demonstrated that follistatin improves in vitro development of embryos produced with sex-sorted semen and such effects are associated with enhanced indices of SMAD signaling.
摘要译文
使用性别分选的精液产生所需性别的后代与体外发育能力降低和体内生育率降低有关。卵泡抑素补充对性别分选精液产生的牛胚胎的发育能力以及TGF-β调节的途径与卵泡抑素的胚胎营养作用之间可能存在联系。研究了卵泡抑素对SMAD信号传导（CTGF，ID1，ID2和ID3）和AKT磷酸化的细胞谱系标志物（CDX2和Nanog）和下游靶标表达的影响。在胚胎培养的最初72小时期间补充卵泡抑素。外源性卵泡抑素将性别分选精液产生的胚胎的体外发育能力恢复到未分选精液产生的对照胚胎水平，使用来自三种不同公牛的分选精液获得了可比较的结果。 SMAD信号传导下游靶基因，CTGF（SMAD 2/3途径）和ID2（SMAD 1/5途径）的mRNA丰度，使用性别分选与未分类精液产生的胚泡中较低，但两组中CDX2，NANOG，ID1和ID3的mRNA水平相似。卵泡抑素补充剂使用性别分选的精液将胚胎中的CTGF和ID2 mRNA恢复至对照胚胎的水平。此外，磷酸化（p）AKT（Ser-473和Thr-308）的水平在来自性别分选和未分选精液的胚胎中相似，但卵泡抑素治疗增加了两组的pAKT水平。总之，结果表明卵泡抑素改善了用性别分选的精液产生的胚胎的体外发育，并且这种效应与SMAD信号传导的增强指数相关。
MohamedAshry[a][b];KyungBonLee[a][c];Joseph K.Folger[a];Sandeep K.Rajput[a][d];George W.Smith[a];. Follistatin supplementation during in vitro embryo culture improves developmental competence of bovine embryos produced using sex-sorted semen[J]. Reproductive Biology, 2018,18(3): 267-273