牛输卵管液中的 miR-17-5p 影响胚胎发育。

miR-17-5p in bovine oviductal fluid affects embryo development.

机构信息

Department of Animal Science, Graduate School of Agriculture, Tokyo University of Agriculture, Funako, 1737, Atsugi City, Kanagawa, Japan.

NODAI Genome Research Center, Tokyo University of Agriculture, Sakuragaoka 1-1-1, Setagaya, Tokyo, Japan.

出版信息

Mol Cell Endocrinol. 2022 Jul 1;551:111651. doi: 10.1016/j.mce.2022.111651. Epub 2022 Apr 20.

DOI:10.1016/j.mce.2022.111651

PMID:35452772

Abstract

This study identified microRNAs (miRNAs) in bovine oviductal fluids (OFs) and examined the effect of miR-17-5p in OFs on embryonic development to the blastocyst stage. Small RNA-seq of extracellular vesicles of OFs revealed 242 miRNAs. Additionally, analyzing expressions of randomly selected OF-miRNAs with RT-qPCR in the culture medium of oviductal epithelial cells indicated that the abundance of miRNAs in OFs increased during the luteal phase. miR-17-5p mimic-treated eight-cell-stage zona pellucida-free embryos showed improved embryonic development to the blastocyst stage. The effect of the miR-17-5p mimic was confirmed using a dual-luciferase assay and immunostaining. In addition, RNA-seq of the miR-17-5p mimic- or control-treated embryos revealed differentially expressed genes (DEGs), suggesting possible pathways that overlapped with the in silico-predicted pathways for miR-17-5p targeting genes. Furthermore, ingenuity pathway analysis of DEG predicted miR-17 to be a significant upstream regulator. Our results suggest that miR-17-5p in OFs regulates embryonic development in bovines.

摘要

本研究鉴定了牛输卵管液（OFs）中的 microRNAs（miRNAs），并研究了 OFs 中的 miR-17-5p 对胚胎发育至囊胚阶段的影响。OFs 细胞外囊泡的小 RNA-seq 显示了 242 种 miRNAs。此外，用 RT-qPCR 分析在输卵管上皮细胞培养基中随机选择的 OF-miRNAs 的表达表明，miRNAs 在 OFs 中的丰度在黄体期增加。miR-17-5p 模拟物处理的无透明带期 8 细胞胚胎显示胚胎发育至囊胚阶段的能力得到改善。通过双荧光素酶测定和免疫染色证实了 miR-17-5p 模拟物的作用。此外，miR-17-5p 模拟物或对照处理胚胎的 RNA-seq 显示差异表达基因（DEGs），提示可能存在与 miR-17-5p 靶基因的计算机预测途径重叠的途径。此外，DEG 的 ingenuity 途径分析预测 miR-17 是一个重要的上游调节剂。我们的结果表明，OFs 中的 miR-17-5p 调节牛胚胎的发育。