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miR-430 对幼稻水蛇类固醇激素合成相关基因表达的综合分析。

Integrated Analysis of miR-430 on Steroidogenesis-Related Gene Expression of Larval Rice Field Eel .

机构信息

College of Fisheries, Huazhong Agricultural University, Wuhan 430070, China.

Engineering Research Center of Green Development for Conventional Aquatic Biological Industry in the Yangtze River Economic Belt, Ministry of Education, Wuhan 430070, China.

出版信息

Int J Mol Sci. 2021 Jun 29;22(13):6994. doi: 10.3390/ijms22136994.

Abstract

The present study aims to reveal the mechanism by which miR-430s regulate steroidogenesis in larval rice field eel . To this end, embryos were respectively microinjected with miRNA-overexpressing mimics (agomir430a, agomir430b, and agomir430c) or miRNA-knockdown inhibitors (antagomir430a, antagomir430b, and antagomir430c). Transcriptome profiling of the larvae indicated that a total of more than 149 differentially expressed genes (DEGs) were identified among the eight treatments. Specifically, DEGs related to steroidogenesis, the GnRH signaling pathway, the erbB signaling pathway, the Wnt signaling pathway, and other pathways were characterized in the transcriptome. We found that steroidogenesis-related genes (hydroxysteroid 17-beta dehydrogenase 3 (), hydroxysteroid 17-beta dehydrogenase 7 (), hydroxysteroid 17-beta dehydrogenase 12 (), and cytochrome P450 family 19 subfamily a ()) were significantly downregulated in miR-430 knockdown groups. The differential expressions of miR-430 in three gonads indicated different roles of three miR-430 (a, b, and c) isoforms in regulating steroidogenesis and sex differentiation. Mutation of the miR-430 sites reversed the downregulation of cytochrome P450 family 17 (), , and forkhead box L2 () reporter activities by miR-430, indicating that miR-430 directly interacted with , , and genes to inhibit their expressions. Combining these findings, we concluded that miR-430 regulated the steroidogenesis and the biosynthesis of steroid hormones by targeting in larval . Our results provide a novel insight into steroidogenesis at the early stage of fish at the molecular level.

摘要

本研究旨在揭示 miR-430s 调节幼稻田鳗类固醇生成的机制。为此,分别向胚胎注射 miRNA 过表达模拟物(agomir430a、agomir430b 和 agomir430c)或 miRNA 敲低抑制剂(antagomir430a、antagomir430b 和 antagomir430c)。幼虫的转录组分析表明,在这 8 种处理中,总共鉴定出超过 149 个差异表达基因(DEGs)。具体而言,转录组中鉴定出与类固醇生成、GnRH 信号通路、erbB 信号通路、Wnt 信号通路和其他通路相关的 DEGs。我们发现,类固醇生成相关基因(羟甾醇 17-β脱氢酶 3 ()、羟甾醇 17-β脱氢酶 7 ()、羟甾醇 17-β脱氢酶 12 ()和细胞色素 P450 家族 19 亚家族 a ())在 miR-430 敲低组中显著下调。三种性腺中 miR-430 的差异表达表明三种 miR-430(a、b 和 c)亚型在调节类固醇生成和性别分化中的不同作用。miR-430 结合位点的突变逆转了 miR-430 对细胞色素 P450 家族 17 ()、和叉头框 L2 ()报告基因活性的下调,表明 miR-430 直接与、和基因相互作用,抑制它们的表达。综合这些发现,我们得出结论,miR-430 通过靶向在幼稻田鳗中调节类固醇生成和类固醇激素的生物合成。我们的研究结果从分子水平为鱼类早期的类固醇生成提供了新的见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6965/8269179/a0aff86b3424/ijms-22-06994-g001.jpg

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