非编码RNA在米非司酮诱导的红鳍东方鲀性腺雄性化中的潜在调控作用。

The potential regulatory role of non-coding RNAs in mifepristone-induced masculinization in Takifugu rubripes gonads.

作者信息

Gao Rui, Yan Hongwei, Zhou Huiting, Hu Mingtao, Wang Jia, Shen Xufang, Li Meiyuan, Chen Jinfeng, Liu Qi, Liu Ying, Wang Xiuli, Sun Qunwen, Zhou He

机构信息

Key Laboratory of Environment Controlled Aquaculture (Dalian Ocean University), Ministry of Education, Beijing, 116023, China.

College of Marine Science and Environment Engineering, Dalian Ocean University, Dalian, Liaoning, 116023, China.

出版信息

BMC Genomics. 2025 May 12;26(1):471. doi: 10.1186/s12864-025-11647-1.

DOI:10.1186/s12864-025-11647-1

PMID:40355821

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12067753/

Abstract

BACKGROUND

The regulatory roles of non-coding RNAs (ncRNAs) during sex differentiation in teleosts have received widespread attention recently. Mifepristone (RU486, a progesterone antagonist), which acts as an endocrine disruptor, can affect reproduction and sex differentiation in teleosts.

RESULTS

The expression of ncRNAs in the gonads of tiger puffer (Takifugu rubripes) during RU486 (500 µg/g diet) induced masculinization process was examined by RNA-sequencing. A total of 4,381 long non-coding RNAs (lncRNAs), 309 circular RNAs (circRNAs), and 1,020 microRNAs (miRNAs) were identified. The expression of 41 differentially expressed (DE) lncRNAs and 20 DE miRNAs, which showed sexual dimorphic expression patterns in genetic female gonads in the control group (C-XX) vs. genetic male gonads in the control group (C-XY), were altered in genetic female gonads in the RU486 treated group (RU-XX). The genes targeted by DE ncRNAs were mainly enriched in sex-related pathways, such as calcium signaling, ovarian steroidogenesis, and cortisol synthesis and secretion. The results of co-expression and competing endogenous RNA (ceRNA) network analysis indicated that miRNAs (e.g., miR-205-z and fru-miR-122) and lncRNAs (including XR_003890915.1 and XR_003885862.1) may have pivotal roles, and lncRNAs (including XR_003890295.1, MSTRG.11750.1, and XR_003888827.1) may act as miRNA sponges, involved in the competition between miRNAs and sex-related genes during tiger puffer masculinization process. Dual luciferase reporter assay results identified that ovarian steroidogenesis related gene hsd17b1 is a downstream target of fru-miR-122. The expression of 4 lncRNAs, 4 circRNAs, and 6 miRNAs were validated by qPCR, indicating the accuracy and dependability of RNA-Seq.

CONCLUSIONS

This study provided the evidence that ncRNAs may participate in RU486-induced masculinization in T. rubripes, and may enhance our understanding of the regulatory network of sex differentiation in fugu.

摘要

背景

非编码RNA（ncRNA）在硬骨鱼性别分化过程中的调控作用近来受到广泛关注。米非司酮（RU486，一种孕酮拮抗剂）作为一种内分泌干扰物，可影响硬骨鱼的繁殖和性别分化。

结果

通过RNA测序检测了在RU486（500μg/g饲料）诱导的雄性化过程中虎河豚（Takifugu rubripes）性腺中ncRNA的表达。共鉴定出4381条长链非编码RNA（lncRNA）、309条环状RNA（circRNA）和1020条微小RNA（miRNA）。在对照组遗传雌性性腺（C-XX）与对照组遗传雄性性腺（C-XY）中表现出性二态性表达模式的41条差异表达（DE）lncRNA和20条DE miRNA的表达，在RU486处理组的遗传雌性性腺（RU-XX）中发生了改变。DE ncRNA靶向的基因主要富集在与性别相关的途径中，如钙信号传导、卵巢类固醇生成以及皮质醇合成与分泌。共表达和竞争性内源RNA（ceRNA）网络分析结果表明，miRNA（如miR-205-z和fru-miR-122）和lncRNA（包括XR_003890915.1和XR_003885862.1）可能起关键作用，lncRNA（包括XR_003890295.1、MSTRG.11750. 和XR_003888827.1）可能作为miRNA海绵，在虎河豚雄性化过程中参与miRNA与性别相关基因之间的竞争。双荧光素酶报告基因检测结果确定卵巢类固醇生成相关基因hsd17b1是fru-miR-122的下游靶标。通过qPCR验证了4条lncRNA、4条circRNA和6条miRNA的表达，表明RNA测序的准确性和可靠性。