Guo Z, Islam M S, Liu D, Liu G, Lv L, Yang Y, Fu B, Wang L, Liu Z, He H, Wu H
Heilongjiang Academy of Agricultural Sciences Postdoctoral Programme, Animal Husbandry Research Institute, Harbin, China.
Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation, Ministry of Agriculture, Beijing, China.
Reprod Domest Anim. 2018 Feb;53(1):3-10. doi: 10.1111/rda.13035. Epub 2017 Nov 14.
Several oocyte-derived genes/proteins are essential to early embryonic development. The expression and stability of these proteins are influenced by the autocrine/paracrine activity of factors released by oocytes and cumulus cells. This study investigated the paracrine and autocrine activity of follistatin (FS), which is secreted by oocytes and cumulus cells as part of porcine embryogenesis. Immunohistochemical (IHC) localization of follistatin was conducted on 100 randomly selected early- and late-cleaving two-cell embryos. Dissociated cumulus cells were treated with various doses of follistatin for determination of the follistatin gene (FST) mRNA expression levels by quantitative real-time PCR analysis. Microinjection of siRNA induced a downregulation of FST mRNA during embryonic development, thereby decreasing the proportion embryos developing to the blastocyst stage (19.33%). Immunolocalization analysis showed enhanced staining for follistatin in early-cleavage stage embryos. Quantitative real-time PCR indicated a significantly lower FST transcript level in cumulus cells after application of the highest dose of follistatin (100 ng/ml). Exogenous follistatin treatment of in vitro maturation embryos resulted in statistically significant dose-dependent changes during development. Application of the highest concentration (100 ng/ml) of follistatin decreased the maturation rate of the oocytes. On the other hand, the application of 10 ng/ml follistatin resulted in an increase in the number of embryos. The observed differential effect of exogenous follistatin might be due to maternal FST and autocrine/paracrine factors secreted by cumulus cells.
几种卵母细胞衍生的基因/蛋白质对早期胚胎发育至关重要。这些蛋白质的表达和稳定性受卵母细胞和颗粒细胞释放因子的自分泌/旁分泌活性影响。本研究调查了卵泡抑素（FS）的旁分泌和自分泌活性，卵泡抑素是猪胚胎发生过程中由卵母细胞和颗粒细胞分泌的。对100个随机选择的早期和晚期分裂的二细胞胚胎进行卵泡抑素的免疫组织化学（IHC）定位。用不同剂量的卵泡抑素处理解离的颗粒细胞，通过定量实时PCR分析测定卵泡抑素基因（FST）mRNA表达水平。显微注射siRNA可在胚胎发育过程中下调FST mRNA，从而降低发育到囊胚阶段的胚胎比例（19.33%）。免疫定位分析显示早期分裂阶段胚胎中卵泡抑素的染色增强。定量实时PCR表明，应用最高剂量的卵泡抑素（100 ng/ml）后，颗粒细胞中FST转录水平显著降低。体外成熟胚胎的外源性卵泡抑素处理在发育过程中导致统计学上显著的剂量依赖性变化。应用最高浓度（100 ng/ml）的卵泡抑素可降低卵母细胞的成熟率。另一方面，应用10 ng/ml卵泡抑素可使胚胎数量增加。观察到的外源性卵泡抑素的差异效应可能归因于母体FST和颗粒细胞分泌的自分泌/旁分泌因子。
