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卵泡液外泌体来源的 miR-339-5p 通过靶向 ERK1/2 通路调节因子 SFPQ 增强猪卵母细胞的体外成熟。

Follicular fluid exosome-derived miR-339-5p enhances in vitro maturation of porcine oocytes via targeting SFPQ, a regulator of the ERK1/2 pathway.

机构信息

Yanbian University, Jilin, Yanji, 133000, China.

Institute of Animal Biotechnology, Jilin Academy of Agricultural Sciences, Jilin, Gongzhuling, 136100, China.

出版信息

Theriogenology. 2024 Sep 1;225:107-118. doi: 10.1016/j.theriogenology.2024.04.022. Epub 2024 May 15.

Abstract

In this study, we aimed to investigate cytoplasmic maturation and miRNA expression of mature oocytes cultured in porcine follicular fluid exosomes. We also examined the effect of miR-339-5p on oocyte maturation. Twenty eight differentially expressed miRNAs were detected using miRNA-seq. We then transfected cumulus oocyte complexes with miR-339-5p mimics and inhibitor during culture. The results showed that exosomes increased endoplasmic reticulum levels and the amount of lipid droplets, and decreased ROS levels, lipid droplet size, and percentage of oocytes with abnormal cortical granule distribution. Overexpressing miR-339-5p significantly decreased cumulus expansion genes, oocyte maturation-related genes, target gene proline/glutamine-rich splicing factor (SFPQ), ERK1/2 phosphorylation levels, oocyte maturation rate, blastocyst rate, and lipid droplet number, but increased lipid droplet size and the ratio of oocytes with abnormal cortical granule distribution. Inhibiting miR-339-5p reversed the decrease observed during overexpression. Mitochondrial membrane potential and ROS levels did not differ significantly between groups. In summary, exosomes promote oocyte cytoplasmic maturation and miR-339-5p regulating ERK1/2 activity through SFPQ expression, thereby elevating oocyte maturation and blastocyst formation rate in vitro.

摘要

在这项研究中,我们旨在研究培养在猪卵泡液外泌体中的成熟卵母细胞的细胞质成熟和 miRNA 表达,并研究 miR-339-5p 对卵母细胞成熟的影响。我们使用 miRNA-seq 检测到 28 个差异表达的 miRNA。然后,在培养过程中用 miR-339-5p 模拟物和抑制剂转染卵丘-卵母细胞复合物。结果表明,外泌体增加了内质网水平和脂滴数量,降低了 ROS 水平、脂滴大小和异常皮质颗粒分布的卵母细胞比例。过表达 miR-339-5p 显著降低了卵丘扩展基因、卵母细胞成熟相关基因、靶基因脯氨酸/谷氨酰胺丰富拼接因子(SFPQ)、ERK1/2 磷酸化水平、卵母细胞成熟率、囊胚率和脂滴数量,但增加了脂滴大小和异常皮质颗粒分布的卵母细胞比例。抑制 miR-339-5p 逆转了过表达观察到的降低。各组间线粒体膜电位和 ROS 水平无显著差异。综上所述,外泌体通过 SFPQ 表达促进卵母细胞细胞质成熟和 miR-339-5p 调节 ERK1/2 活性,从而提高体外卵母细胞成熟和囊胚形成率。

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