University of Wisconsin, Department of Animal Sciences, Madison, WI, 53706, USA.
University of Milan, Department of Veterinary Medicine, Lodi, 26900, Italy.
Sci Rep. 2019 Dec 30;9(1):20347. doi: 10.1038/s41598-019-55868-3.
Embryos utilize oocyte-donated RNAs until they become capable of producing RNAs through embryonic genome activation (EGA). The sperm's influence over pre-EGA RNA content of embryos remains unknown. Recent studies have revealed that sperm donate non-genomic components upon fertilization. Thus, sperm may also contribute to RNA presence in pre-EGA embryos. The first objective of this study was to investigate whether male fertility status is associated with the RNAs present in the bovine embryo prior to EGA. A total of 65 RNAs were found to be differentially expressed between 2-4 cell bovine embryos derived from high and low fertility sires. Expression patterns were confirmed for protein phosphatase 1 regulatory subunit 36 (PPP1R36) and ataxin 2 like (ATXN2L) in three new biological replicates. The knockdown of ATXN2L led to a 22.9% increase in blastocyst development. The second objective of this study was to characterize the parental origin of RNAs present in pre-EGA embryos. Results revealed 472 sperm-derived RNAs, 2575 oocyte-derived RNAs, 2675 RNAs derived from both sperm and oocytes, and 663 embryo-exclusive RNAs. This study uncovers an association of male fertility with developmentally impactful RNAs in 2-4 cell embryos. This study also provides an initial characterization of paternally-contributed RNAs to pre-EGA embryos. Furthermore, a subset of 2-4 cell embryo-specific RNAs was identified.
胚胎利用卵母细胞捐赠的 RNA，直到它们能够通过胚胎基因组激活（EGA）产生 RNA。精子对 EGA 前胚胎 RNA 含量的影响尚不清楚。最近的研究表明，精子在受精时会捐赠非基因组成分。因此，精子也可能有助于 EGA 前胚胎中 RNA 的存在。本研究的第一个目的是研究男性生育能力是否与 EGA 前牛胚胎中存在的 RNA 有关。发现 65 种 RNA 在高、低生育能力公牛来源的 2-4 细胞牛胚胎中存在差异表达。在三个新的生物学重复中，对蛋白磷酸酶 1 调节亚基 36（PPP1R36）和共济失调 2 样（ATXN2L）的表达模式进行了确认。ATXN2L 的敲低导致囊胚发育增加了 22.9%。本研究的第二个目的是表征 EGA 前胚胎中存在的亲本来源的 RNA。结果显示了 472 种精子来源的 RNA、2575 种卵母细胞来源的 RNA、2675 种来自精子和卵母细胞的 RNA 以及 663 种胚胎特有的 RNA。本研究揭示了男性生育能力与 2-4 细胞胚胎中具有发育影响的 RNA 之间的关联。本研究还首次对 EGA 前胚胎中父本贡献的 RNA 进行了特征描述。此外，还鉴定了一组 2-4 细胞胚胎特异性 RNA。
