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miR-450-5p and miR-202-5p Synergistically Regulate Follicle Development in Black Goat

文献类型： 外文期刊

作者： Feng, Guanghang 1 ; Liu, Jie 1 ; Lu, Zitao 1 ; Li, Yaokun 1 ; Deng, Ming 1 ; Liu, Guangbin 1 ; Sun, Baoli 1 ; Guo, Yongqing 1 ; Zou, Xian 2 ; Liu, Dewu 1 ;

作者机构： 1.South China Agr Univ, Coll Anim Sci, Herbivore Lab, Guangzhou 510642, Peoples R China

2.Guangdong Acad Agr Sci, Inst Anim Sci, State Key Lab Livestock & Poultry Breeding, Guangdong Key Lab Anim Breeding & Nutr, Guangzhou 510640, Peoples R China

3.South China Agr Univ, Natl Joint Engn Res Ctr, Guangzhou 510642, Peoples R China

4.South China Agr Univ, Guangdong Key Lab Agr Anim Genom & Mol Breeding, Guangzhou 510642, Peoples R China

5.Shihezi Univ, Collaborat Innovat Ctr Hlth Sheep Breeding & Zoono, Shihezi 832000, Peoples R China

关键词： goat; follicle development; granulosa cells; MiR-202-5p; MiR-450-5p

期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.6； 五年影响因子：6.2 ）

ISSN：

年卷期： 2023 年 24 卷 1 期

页码：

收录情况： SCI

摘要： Follicle maturation is a complex biological process governed by numerous factors, and researchers have observed follicle development by studying the proliferation and apoptosis of follicular granulosa cells (GCs). However, the regulatory mechanisms of GCs proliferation and death during follicle development are largely unknown. To investigate the regulatory mechanisms of lncRNAs, mRNAs, and microRNAs, RNA sequencing (RNA-seq) and small RNA-seq were performed on large (>10 mm) and small follicles (<3 mm) of Leizhou black goat during estrus. We discovered two microRNAs, miR-450-5p and miR-202-5p, which can target GCs in goats and may be involved in follicle maturation, and the effects of miR-450-5p and miR-202-5p on ovarian granulosa cell lines were investigated (KGN). Using cell counting kit-8 (CCK-8) assays, 5-Ethynyl-2'-deoxyuridine (EdU) assay and flow cytometry, miR-202-5p overexpression could suppress the proliferation and induce apoptosis of GCs, whereas miR-450-5p overexpression induced the opposite effects. The dual-luciferase reporter assay confirmed that miR-450-5p could directly target the BMF gene (a BCL2 modifying factor), and miR-202-5p targeted the BCL2 gene. A considerable rise in phosphorylated Akt (p-AKT) protein was observed following the downregulation of BMF by miR-450-5p mimics. After BMF gene RNAi therapy, a notable elevation in p-AKT was detected. Mimics of miR-202-5p inhibited BCL2 protein expression, significantly decreasing p-AMPK protein expression. These results imply that during the follicular development in black goats, the miR-450-5p-BMF axis favored GC proliferation on a wide scale, while the miR-202-5p-BCL2 axis triggered GC apoptosis.

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