Domingues William B, Silveira Tony L R, Nunes Leandro S, Blodorn Eduardo B, Schneider Augusto, Corcine Carine D, Varela Junior Antônio S, Acosta Izani B, Kütter Mateus T, Greif Gonzalo, Robello Carlos, Pinhal Danillo, Marins Luís F, Campos Vinicius F
Laboratório de Genômica Estrutural, Programa de Pós-Graduação em Biotecnologia, Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, Brazil.
Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande, Rio Grande, Brazil.
Front Genet. 2021 Sep 8;12:704778. doi: 10.3389/fgene.2021.704778. eCollection 2021.
Overexpression of growth hormone (GH) in -transgenic zebrafish of a highly studied lineage F0104 has earlier been reported to cause increased muscle growth. In addition to this, GH affects a broad range of cellular processes in transgenic fish, such as morphology, physiology, and behavior. Reports show changes such as decreased sperm quality and reduced reproductive performance in transgenic males. It is hypothesized that microRNAs are directly involved in the regulation of fertility potential during spermatogenesis. The primary aim of our study was to verify whether overexpression disturbs the sperm miRNA profile and influences the sperm quality in transgenic zebrafish. We report a significant increase in body weight of -transgenic males along with associated reduced sperm motility and other kinetic parameters in comparison to the non-transgenic group. MicroRNA transcriptome sequencing of -transgenic zebrafish sperms revealed expressions of 186 miRNAs, among which six miRNA were up-regulated (miR-146b, miR-200a-5p, miR-146a, miR-726, miR-184, and miR-738) and sixteen were down-regulated (miR-19d-3p, miR-126a-5p, miR-126b-5p, miR-22a-5p, miR-16c-5p, miR-20a-5p, miR-126b-3p, miR-107a-3p, miR-93, miR-2189, miR-202-5p, miR-221-3p, miR-125a, miR-125b-5p, miR-126a-3p, and miR-30c-5p) in comparison to non-transgenic zebrafish. Some of the dysregulated miRNAs were previously reported to be related to abnormalities in sperm quality and reduced reproduction ability in other species. In this study, an average of 134 differentially expressed miRNAs-targeted genes were predicted using the approach. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis demonstrated that the genes of affected pathways were primarily related to spermatogenesis, sperm motility, and cell apoptosis. Our results suggested that excess GH caused a detrimental effect on sperm microRNAome, consequently reducing the sperm quality and reproductive potential of zebrafish males.
在一个经过深入研究的品系F0104的转基因斑马鱼中，生长激素（GH）的过表达此前已被报道会导致肌肉生长增加。除此之外，GH还会影响转基因鱼的一系列细胞过程，如形态、生理和行为。报告显示转基因雄性鱼存在精子质量下降和繁殖性能降低等变化。据推测，微小RNA直接参与精子发生过程中生育潜力的调节。我们研究的主要目的是验证过表达是否会扰乱转基因斑马鱼的精子微小RNA谱并影响精子质量。我们报告称，与非转基因组相比，转基因雄性鱼的体重显著增加，同时精子活力及其他动力学参数降低。对转基因斑马鱼精子进行的微小RNA转录组测序揭示了186种微小RNA的表达，其中6种微小RNA上调（miR-146b、miR-200a-5p、miR-146a、miR-726、miR-184和miR-738），16种下调（miR-19d-3p、miR-126a-5p、miR-126b-5p、miR-22a-5p、miR-16c-5p、miR-20a-5p、miR-126b-3p、miR-107a-3p、miR-93、miR-2189、miR-202-5p、miR-221-3p、miR-125a、miR-125b-5p、miR-126a-3p和miR-30c-5p）。一些失调的微小RNA此前在其他物种中被报道与精子质量异常和繁殖能力降低有关。在本研究中，使用该方法预测了平均134个差异表达的微小RNA靶向基因。京都基因与基因组百科全书（KEGG）通路富集分析表明，受影响通路的基因主要与精子发生、精子活力和细胞凋亡有关。我们的结果表明，过量的GH对精子微小RNA组产生了有害影响，从而降低了斑马鱼雄性的精子质量和繁殖潜力。
