Houdelet Camille, Blondeau-Bidet Eva, Estevez-Villar Mathilde, Mialhe Xavier, Hermet Sophie, Ruelle François, Dutto Gilbert, Bajek Aline, Bobe Julien, Geffroy Benjamin
MARBEC Univ Montpellier, CNRS, Ifremer, IRD, Palavas-Les-Flots, France.
MARBEC Univ Montpellier, CNRS, Ifremer, IRD, Montpellier, France.
Mar Biotechnol (NY). 2023 Oct;25(5):749-762. doi: 10.1007/s10126-023-10237-0. Epub 2023 Aug 15.
MicroRNAs (miRNAs) constitute a new category of biomarkers. Studies on miRNAs in non-mammalian species have drastically increased in the last few years. Here, we explored the use of miRNAs as potential, poorly invasive markers, to identify sex and characterize acute stress in fish. The European seabass (Dicentrarchus labrax) was chosen as a model because of its rapid response to stress and its specific sex determination system, devoid of sexual chromosomes. We performed a small RNA-sequencing analysis in the blood plasma of male and female European seabass (mature and immature) as well as in the blood plasma of juveniles submitted to an acute stress and sampled throughout the recovery period (at 0 h, 0.5 h, 1.5 h and 6 h). In immature individuals, both miR-1388-3p and miR-7132a-5p were up-regulated in females, while miR-499a-5p was more abundant in males. However, no miRNAs were found to be differentially expressed between sexes in the blood plasma of mature individuals. For the acute stress analysis, five miRNAs (miR-155-5p, miR-200a-3p, miR-205-1-5p, miR-143-3p, and miR-223-3p) followed cortisol production over time. All miRNAs identified were tested and validated by RT-qPCR on sequenced samples. A complementary analysis on the 3'UTR sequences of the European seabass allowed to predict potential mRNA targets, some of them being particularly relevant regarding stress regulation, e.g., the glucocorticoid receptor 1 and the mineralocorticoid receptor. The present study provides new avenues and recommendations on the use of miRNAs as biomarkers of sex or stress of the European seabass, with potential application on other fish species.
微小RNA（miRNA）构成了一类新的生物标志物。在过去几年中，对非哺乳动物物种中miRNA的研究急剧增加。在此，我们探索了将miRNA用作潜在的、低侵入性标志物，以识别鱼类的性别并表征其急性应激反应。由于欧洲海鲈（Dicentrarchus labrax）对压力反应迅速且具有独特的性别决定系统（无性染色体），因此被选为模型。我们对雄性和雌性欧洲海鲈（成熟和未成熟）的血浆以及遭受急性应激并在整个恢复期（0小时、0.5小时、1.5小时和6小时）采样的幼鱼血浆进行了小RNA测序分析。在未成熟个体中，雌性中miR-1388-3p和miR-7132a-5p均上调，而miR-499a-5p在雄性中更为丰富。然而，在成熟个体的血浆中未发现有性别差异表达的miRNA。对于急性应激分析，五种miRNA（miR-155-5p、miR-200a-3p、miR-205-1-5p、miR-143-3p和miR-223-3p）随时间跟踪皮质醇的产生。对所有鉴定出的miRNA在测序样本上通过RT-qPCR进行了测试和验证。对欧洲海鲈3'UTR序列的补充分析能够预测潜在的mRNA靶标，其中一些在应激调节方面特别相关，例如糖皮质激素受体1和盐皮质激素受体。本研究为将miRNA用作欧洲海鲈性别或应激的生物标志物提供了新途径和建议，并有可能应用于其他鱼类物种。
