Sexual Dimorphism of miRNAs Secreted by Bovine In vitro-produced Embryos.
Sexual Dimorphism of miRNAs Secreted by Bovine In vitro-produced Embryos.
复制标题
DOI：
10.3389/fgene.2017.00039
复制
发表时间：
2017
影响因子：
3.7
通讯作者：
Khatib H
中科院分区：
文献类型：
--
作者：
Gross N;Kropp J;Khatib H
牛囊胚的性别二态性此前已通过发育、细胞死亡、代谢、端粒长度、DNA 甲基化和转录组学的差异观察到。然而，miRNA 分泌到培养基中的二态性尚未得到评估。本研究的目的是确定是否发生性别特异性囊胚 miRNA 分泌，并进一步研究这些 miRNA 在囊胚与母体环境之间的相互作用中可能发挥的作用。进行体外胚胎培养，并将来自雄性和雌性囊胚的培养基收集到性别特异性池中。对 68 个 miRNA 的分析揭示了总共 8 个 miRNA 在雌性和雄性条件培养基中存在差异表达。 qPCR 验证证实，在另外三个生物学重复中，女性培养基中 miR-22 (P < 0.05)、miR-122 (P < 0.05) 和 miR-320a (P < 0.05) 的表达较高。为了检查分泌的 miRNA 在与母体环境通讯中对介质的潜在作用，将 miR-22、miR-122 和 miR-320a 分别补充到原代牛子宫内膜上皮细胞培养物的四个重复中。检测miR-122（P < 0.05）和miR-320a（P < 0.05）的摄取，并检测miR-22的摄取趋势（P > 0.05）。此外，发现补充 miR-122 (P < 0.05) 和 miR-320a (P < 0.05) 后，黄体酮受体转录物（所有三种 miRNA 的预测靶标）的表达在细胞中上调，并且在补充 miR-22 (P > 0.05) 后观察到转录物的趋势上调。这项工作表明，雄性和雌性受孕者能够在囊胚阶段差异性地分泌 miRNA，并且这些 miRNA 在应用于母体细胞时能够诱导转录组反应。这一知识建立在囊胚阶段概念的已知二态性差异的基础上，并证明了囊胚分泌的 miRNA 在细胞间通讯中的作用。
Sexual dimorphism of bovine blastocysts has previously been observed through differences in development, cell death, metabolism, telomere length, DNA methylation, and transcriptomics. However, dimorphism in the secretion of miRNAs to culture media has not yet been evaluated. The objectives of this study were to determine if sex-specific blastocyst miRNA secretion occurs and to further investigate the role these miRNAs may have in the interaction between a blastocyst and the maternal environment. In vitro embryo culture was performed and media from male and female blastocysts was collected into sex-specific pools. Profiling of 68 miRNAs revealed a total of eight miRNAs that were differentially expressed between female and male-conditioned media. Validation by qPCR confirmed higher expression of miR-22 (P < 0.05), miR-122 (P < 0.05), and miR-320a (P < 0.05) in female media for three additional biological replicates. To examine the potential roles of secreted miRNAs to the media in communication with the maternal environment, miR-22, miR-122, and miR-320a were each supplemented to four replicates of primary bovine endometrial epithelial cell culture. Uptake of miR-122 (P < 0.05) and miR-320a (P < 0.05) was detected, and a trend of uptake was detected for miR-22 (P > 0.05). Further, expression of the progesterone receptor transcript, a predicted target of all three miRNAs, was found to be upregulated in the cells following supplementation of miR-122 (P < 0.05) and miR-320a (P < 0.05), and a trend upregulation of the transcript was observed following miR-22 (P > 0.05) supplementation. This work demonstrates that male and female conceptuses are able to differentially secrete miRNAs at the blastocyst stage and that these miRNAs have the ability to induce a transcriptomic response when applied to maternal cells. This knowledge builds on the known dimorphic differences in conceptuses at the blastocyst stage and demonstrates a role for blastocyst-secreted miRNAs in cell–cell communication.